bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunophenotypic Diagnostic Score for MDS Taking into Account Various Cell Populations.
Authors: Millaret Arnaud , Galmarini Carlos , Ritouet Danielle Treille , Gerland Luc M. , Dumontet Charles M. 
Published: October 2019

Abstract
Immunophenotyping has proven to be an interesting approach in patients with myelodysplastic syndromes (MDS)(Maynadie et al., Blood. 2002 Oct 1;100(7):2349–56). Most studies in these diseases have focused on the analysis of blast cells. We chose to determine the potential interest of markers expressed on other bone marrow compartments, namely granulocytes, monocytes and the CD34 compartment.
Fourteen marrow samples from patients with MDS and 10 normal bone marrow were analyzed. The median age in the MDS group and the control group was 64.4 and 59.8 years, respectively. Flow cytometry analysis of bone marrow samples was performed on a Facs Calibur, using four-color panels. The following antigens were analyzed: CD7, CD11c, CD14, CD15, CD16, CD33, CD34, CD45, CD36, CD38, CD56, CD71, CD117 and HLA-DR. The various compartments analyzed were defined using CD45/SSC gating, combined with SSC/CD34 gating for CD34+ cells. The median percentage of blasts in the MDS marrows was < 5%. Seven patients had a hemoglobin value < 100 g/dl. Five patients had a leukocyte count < 3.109/l. Six patients had a platelet count < 100.1012/L. Threshold values were determined by ROC analysis. Comparison of the expression levels of markers between MDS and normal bone marrows was performed using a Wilcoxon test. Analysis was performed on bone marrow granulocytes, monocytes and CD34 cells.
Granulocytes were found to express higher levels of CD33 in MDS than in normal marrows (p=0.007). CD34 + cells in MDS marrows expressed significantly higher levels of CD11c (p=0.007), CD117 (p=0.023) and CD33 (p=0.013) than their normal counterparts, with a trend towards lower expression of CD71 (p=0.053). Monocytes in MDS marrows expressed significantly lower levels of CD16 (p=0.006), CD36 (p=0.004) and CD117 (p=0.007) than their normal counterparts, with a trend towards lower expression of CD11c (p=0.059). We did not observe any differences in the antigen expression of MDS erythroblasts in comparison to their normal counterparts. A diagnostic score (0–7) was constructed taking into account the following seven parameters: expression of CD33 on granulocytes; CD11c, CD33 and CD117 on CD34+ cells; CD16, CD36 and CD117 on monocytes. This score distinguished all cases of MDS samples from controls. These results suggest that analysis of cell populations other than blasts could be useful in patients with MDS and that an immunophenotypic panel composed of markers commonly used in the diagnosis of acute leukemias could constitute a useful tool, in complement to morphological studies, for the diagnosis of myelodysplastic syndromes.
Management of Patients with Multiple Myeloma (MM) Failing Total Therapy 2 (TT 2) According to Thalidomide (THAL) Randomization.
Authors: Barlogie Bart , Rasmussen Erik , Tricot Guido , Crowley John 
Published: October 2019

Abstract
As of August 04, 104 of 668 patients enrolled in TT 2 have relapsed/progressed, 61 of whom had been randomized to no THAL and 43 to THAL. The 2 groups were comparable in 14 pre-treatment features examined including the presence of cytogenetic abnormalities (CA), LDH, B2M and albumin. At the time of treatment failure, which occurred at a similar median time from study enrollment of 23 months, higher proportions of patients on THAL had CA (63% vs 37%, p=.03) and higher levels of bone marrow plasmacytosis >33% (46% vs 12%, p=.001). Their subsequent management included THAL + DEX in 25%; Velcade + THAL in 16% and DT-PACE in 10% of those failing on the no THAL arm; compared to THAL + DEX in 12%; Velcade + THAL in 14% and DT-PACE in 21% of those relapsing on the THAL arm. Subsequent salvage response was 51% for those relapsing on no THAL and 26% of those on THAL (relapsing on THAL) (p=.028). Importantly, survival from the time of TT 2 failure was significantly better when patients had not received prior THAL than when THAL was part of the initial management (median of 29 vs 8 mos, p=.0001).
Univarately adverse features for post-failure survival included the presence of CA at the time of failure, IgA isotype, LDH at baseline prior to TT 2 and initial randomization to THAL. On multivariate analysis, baseline LDH (HR 2.6, p=.0006) and THAL randomization (HR 2.6, p=.0006) were highly associated with inferior post-TT 2 failure survival (R2 =38%). Indeed, those on the THAL arm had inferior survival regardless of CA, present either at baseline or at relapse (Figure 1). Our preliminary conclusion is that initial use of THAL for the primary management of symptomatic MM reduces the subsequent salvage potential, due to the emergence of a more aggressive disease exhibiting CA at relapse and greater tumor burden (high bone marrow plasmacytosis). Thus, randomized trials are needed to determine whether thalidomide is more appropriately used up front or for the management of relapse in order to achieve the longest overall survival.
Figure Figure
Imatinib Mesylate in the Treatment of Systemic Mastocytosis, a Phase I/II Trial.
Authors: Droogendijk H.J. , Kluin-Nelemans J.C. , van Daele P.L.A 
Published: October 2019

Abstract
Introduction: mastocytosis comprimes a group of diseases characterized by abnormal proliferation and accumulation of mast cells in one or more organs. A cutaneous and systemic form of mastocytosis is distinguished. Systemic mastocytosis defines the disease process in which mast cell proliferation exceeds the skin. The clinical manifestations of systemic mastocytosis depend on the tissues involved and the tissue response to the accumulation of mast cells. Although in general the disease progresses slowly, it may develop into a malignant disease. Currently there is no cure for systemic mastocytosis. Mast cells develop from pluripotent bone marrow progenitor cells that express CD34 antigen and are dispersed as precursors which undergo proliferation and maturation in different tissues. Normal mast cell development involves the action of stam cell growth factor and c-kit receptors, which are expressed by mast cells at their different developmental stages. Deregulation and/or abnormalities of the c-kit receptor are assumed to play a causal role in disordered mast-cell proliferation. In most patients a mutation in the gene for c-kit exists. One of the mutations is the D816V mutation.
Aim of the study:imatinib mesylate, formerly called ST1571, is a potent inhibitor of c-kit receptor tyrosine kinase activity. In this study, we evaluate whether imatinib mesylate is safe and effective in the treatment of patients with systemic mastocytosis. Primary end-points of study are reduction in urinary N-methylhistamine excretion, serum tryptase activity, skin lesions, number of mast cells in sections of bone marrow, hepato-and/or splenomegaly and symptoms.Adverse effects on therapy are also considered.
Results: up to now, 10 patients with systemic mastocytosis are treated with 400 mg of imatinib mesylate orally once daily. During the first 2 weeks of the study the patients also received 30 mg of prednisolone daily. In general imatinib mesylate is well tolerated. The first results show a 38–80% reduction in urinary N-methylhistamine excretion and 30–66% reduction in serum tryptase activity. Skin lesions diminish in two of the six patients with cutaneous mastocytosis,. Number of mast cells in sections of bone marrow are reduced in 63% (5/8) of the patients. Hepato-and/or splenomegaly is slightly decreased in two of the three patients with organomegaly. Finally 60 % of all patients experiences relief of symptoms. In eight patients the D816V mutation was found. In contrast with former studies imatinib mesylate is also effective in these patients. Further results are to be awaited.
Conclusion: imatinib mesylate is safe and seems effective in the treatment of patients with systemic mastocytosis (including patients with the D816V mutation).
Enhanced Apoptosis to Chemotherapeutic Agents Is Dependent on NFκB and Bcl2-Related Proteins but Is Independent of P53 and Bax in Burkitt’s Lymphoma Cells.
Authors: Kanda Kayoko , Wong Wendy , Boxer Linda M. 
Published: October 2019

Abstract
Deregulated expression of c-Myc is involved in the pathogenesis and growth of Burkitt’s lymphoma cells. Aberrant NFκB activation has been linked to several forms of cancers, including Hodgkin’s lymphoma, breast cancers, and B-cell malignancies. The cytoplasmic retention of NFκB is mediated by a family of inhibitor proteins, the IκBs. We showed that expression of a super-repressor form of IκBα, IκBα-SR, resulted in decreased proliferation and an increased G0/G1 population of Raji Burkitt’s cells. No spontaneous apoptosis was observed, but there was increased sensitivity to apoptosis following treatment with chemotherapeutic agents. Further studies were performed to determine the mechanisms involved in the induction of apoptosis. Interference with NFκB activity decreased the levels of c-Myc and Bcl-xL, but there was no change in the levels of Bax, Bak, Mcl-1, or p53. Treatment with chemotherapeutic agents alone resulted in decreased expression of Mcl-1 with no change in the other proteins measured. Decreased expression of both Bcl-xL and Mcl-1 following inhibition of NFκB activity and chemotherapy (vincristine, 4HC or doxorubicin) treatment resulted in apoptosis. In support of the important role of Bcl-xL and Mcl-1, siRNA experiments revealed that decreased expression of either one alone did not lead to apoptosis, while decreased expression of both Bcl-xL and Mcl-1 resulted in the induction of apoptosis. Many Burkitt’s lymphoma cells have mutations in p53. We found that the mutated p53 was able to bind to IκBα and Iκ Bα-SR as well as to Bcl-xL, however, no up-regulation of Bax or p53 was observed in response to a chemotherapeutic agent. This indicates that mutated p53 retains the ability to bind the other proteins but has lost transcriptional function. Introduction of high levels of wild-type p53 into Raji Burkitt’s cells was able to restore the induction of Bax and Mdm2 and induce apoptosis, whereas the mutant was not. These results suggest that sensitization of Burkitt’s cells to apoptotic death with chemotherapeutic agents requires interference with NFκB activity and changes in the expression of Bcl2-family members but is independent of p53. Inhibition of NFκB activity could be useful in combination with chemotherapy for the treatment of Burkitt’s lymphoma.
Contig Array CGH at 3p14.2 Points to the FHIT Gene as the Deleted Gene in Diffuse Large B Cell Lymphoma.
Authors: Kameoka Yoshihiro , Tagawa Hiroyuki , Tsuzuki Shinobu , Ota Akinobu , Suzuki Ritsuro , Seto Masao 
Published: October 2019

Abstract
Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin’s lymphoma (NHL), accounting for 30 – 40% of adult NHL. Genomic alterations in DLBCL have been investigated by various methods and many regions of amplifications and losses have been detected. Genomic deletions of the 3p arm have been reported in many solid tumors and leukemias, but no study published to date has reported genomic deletions of this region in DLBCL. Recently we demonstrated that 3p14.2 was deleted in approximately 30% of DLBCL patients and cell lines by use of a genome-wide array-comparative genomic hybridization (array-CGH) (Tagawa et al., 2004). For a more detailed examination of the genomic losses at 3p14.2, here we made use of contig BAC array for 3p14.2, and found that 12 of 27 DLBCL samples displayed losses. All of the deleted regions were located within the Fragile Histidine Triad (FHIT) gene, and the most frequently region of loss was mapped to 0.4 Mbp of the region encompassing the introns 4 and 5 and exon 5 of the FHIT gene. Genomic deletions of the FHIT gene have been observed in most common forms of cancer and it is therefore conceivable that the FHIT gene is the target one of the genomic deletion at 3p in DLBCL. To investigate the relationship between genomic deletions and transcript alterations, we performed nested reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Aberrant transcripts or loss of expression was detected in 33% (19 of 57) of the DLBCL samples, and the lost exons of the aberrant transcripts were correlated with genomic deletions detected by array CGH. We also investigated the CpG island methylation status of the promotor of the FHIT gene by means of methylation specific PCR (MSP). One case with genomic loss of the FHIT gene, and three cases without genomic loss showed methylated allele. These findings indicate that 1) Loss of genomic material at 3q14.2 is responsible for exon losses of the FHIT gene, and 2) Genomic loss of the FHIT gene is one of the causes of the generation of aberrant transcripts. 3) Both genomic deletions and methylations are involved in the causes of inactivation of the FHIT gene.
Direct Adoptive Transfer of Cytomegalovirus-Specific CTL from Unrelated Donors to Stem Cell Transplant Patients Following Selection by HLA-Peptide Tetramers.
Authors: Cobbold Mark , Amrolia Persis , Mahendra Prem , Steward Colin , Olavarria Eduardo , Arrazi Julie , McDonald Dorothy , Tauro Sudhir , Goldman John , Craddock Charles , Moss Paul 
Published: October 2019

Abstract
Cytomegalovirus (CMV) infection remains an important cause of morbidity and mortality in patients undergoing allogeneic stem cell transplantation (SCT). This is particularly true in patients transplanted using volunteer unrelated donors (VUDs) where despite improvements in antiviral therapy CMV positive serostatus remains an adverse prognostic indicator. Adoptive transfer of donor-derived CMV-specific CD8+ T cell clones reduces the rate of viral reactivation but the complexity of this approach severely limits its clinical application.
We have previously reported the purification of CMV-specific CD8+ cytotoxic T lymphocytes (CMV-CTL) from the blood of stem cell transplant donors using HLA-peptide tetramers and their subsequent transfer. Six patients undergoing stem cell transplantation from HLA identical sibling donors were studied. CMV-CTL were infused on a pre-emptive basis in patients who developed evidence of CMV viremia. All patients showed prompt and durable CMV-specific immune reconstitution demonstrating durable CMV-specific immune reconstitution. CMV-CTL constituted up to 21% of the patient lymphocyte CD8+ T cell compartment and were detectable up to 24 months in all patients. There was a reduced requirement for anti-viral agents in treated patients (33%) compared with a control group (82%) p=0.11. No secondary reactivation was seen in any patient in the treatment group. No infusion-related toxicity was seen in any treated patients. We have now extended these studies to recipients of VUD transplants in whom no CMV-CTL are detectable within the first 100 days of stem cell infusion. Tetramer-selected CMV-CTL have been adoptively transferred into six patients undergoing transplantation from an unrelated donor. CMV-CTL were given either for treatment of refractory CMV infection (n=5) or prophylactically 21 days post SCT (n=1). A cell dose of between 6x102/kg and 1.25x105/kg was administered with median purity of 95% of the infused T cell dose. No infusional toxicity or acute graft-versus-host disease was noted. CMV-CTL were not present in any patient prior to therapy. Within 7 days of adoptive transfer CMV-CTL were detectable in all patients comprising 0.6% – 20% of CD8+ T cell compartment. All patients demonstrated a reduction in the viral load and 5/6 cleared their viremia. In the patient who received prophylactic therapy durable expansion of CMV-CTL were observed up to 20% of CD8+ T cell pool. Taken together these data are consistent with the hypothesis that antigen-specific CMV specific CTL can be safely infused to recipients of VUD SCT and expand directly in vivo. Such an approach has considerable potential in the treatment or prophylaxis of CMV infection in patients undergoing VUD SCT.
G12/13 Pathways Potentiate Akt Phosphorylation in Platelets Mediated by Gi/Gz Pathways in a Src Kinase-Dependent Manner.
Authors: Kim Soochong , Jin Jianguo , Kunapuli Satya P. 
Published: October 2019

Akt is a serine/threonine kinase that is activated by various agonists including thrombin and ADP in platelets, and activation of Akt in platelets is known to require Gi signaling pathways. Even though thrombin-induced Akt phosphorylation depends on secretion/Gi pathways, thrombin caused much stronger Akt phosphorylation than ADP and epinephrine. In this study, we investigated the contribution of G12/13 pathways to Akt phosphorylation mediated by Gi or Gz pathways. We used selective agonists to activate different G protein pathways. PAR4-activating peptide (AYPGKF) and thrombin failed to induce Akt phosphorylation in Gαq-deficient platelets, but Akt phosphorylation was restored to the levels achieved by AYPGKF and thrombin in wild-type platelets by selective supplement of either Gi or Gz signaling with 2-MeSADP and epinephrine, respectively. This phosphorylation of Akt was dramatically inhibited in the presence of PP2
(4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazole[3,4-d]pyrimidine), an inhibitor of Src family tyrosine kinase, but not by PP3, an inactive structural analog. Importantly, AYPGKF and thrombin induced the activation of Src kinase in Gαq-deficient platelets, suggesting the involvement of Src-dependent pathways in the G12/13 signaling in potentiation of Akt phosphorylation. When human platelets were stimulated with low concentrations of YFLLRNP, a PAR1-specific partial agonist, to selectively activate the G12/13 signaling cascade, Akt phosphorylation did not occur. However, combined stimulation of YFLLRNP-mediated G12/13 signaling and selective activation of Gi pathways caused the Akt phosphorylation. This Akt phosphorylation was blocked by the P2Y12 receptor antagonist AR-C69931MX, or the PI 3-kinase inhibitor LY294002. Platelet aggregation induced by co-activation of both G12/13 and Gi signaling was dramatically inhibited by ML-9
(1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride), an Akt selective inhibitor, suggesting an important role of Akt in platelet aggregation stimulated by combined G12/13 and Gi pathways. These data demonstrate that G12/13 signaling alone does not cause Akt phosphorylation in platelets, but G12/13 signaling has a significant role in potentiating Akt phosphorylation mediated by selective Gi or Gz signaling in human as well as mouse platelets. Finally, we conclude that Src family kinases play an important role in this Akt phosphorylation in platelets.
Red Blood Cell S-Nitrosohemoglobin Deficiency in Pulmonary Arterial Hypertension.
Authors: Pawloski John R. , McMahon Timothy J. , Moya Martin P. , Gow Andrew J. , Huang Yuh-Chin T. , Luchsinger Benjamin J. , Krichman Abigail D. , Bashore Thomas M. , Califf Robert M. , Singel David J. , Piantadosi Claude A. , Tapson Victor F. , Stamler Jonathan S. 
Published: October 2019

Abstract
Recent studies have suggested that binding of oxygen to hemoglobin (Hb) facilitates reactions of nitric oxide (NO) that lead to production of S-nitrosohemoglobin (SNO-Hb), and that vasodilator S-nitrosthiol (SNO) is dispensed by red blood cells (RBCs) at low oxygen tension (pO2) to dilate blood vessels. In human lungs, NO bioactivity serves to attenuate hypoxic pulmonary vasoconstriction (HPV). We therefore considered the possibility that RBC-SNO may oppose HPV and that defective vasodilation by RBCs may contribute to the pathophysiology of pulmonary arterial hypertension (PAH). Here we report that RBCs from patients with PAH exhibit substantial depletion of SNO-Hb and consequent impairment in hypoxia-mediated vasodilation. Furthermore, levels of RBC-NO correlated inversely with pulmonary artery pressures. A SNO-Hb deficiency characteristic of PAH was reproduced in control RBCs by hypoxia: loss of SNO-Hb was accompanied by a buildup of heme-NO species that are deficient in the pO2-governed intramolecular transfer of NO to cysteine thiol, yielding RBCs deficient in NO bioactivity. SNO-deficient RBCs produced exaggerated HPV responses as compared to SNO-replete RBCs. In PAH patients, SNO-Hb repletion fully restored the hypoxic vasodilator activity of RBCs. Our results suggest that a deficiency in RBC-SNO contributes to pulmonary hypertension and hypoxemia, and that repletion of RBC-SNO represents a rational strategy for treating PAH patients.
TAL1 Regulates Expression of the Band 3 Gene in Erythroid Progenitors through Both Upstream Regulatory and Proximal Promoter Elements.
Authors: Xie Jingping , Xu Zhixiong , Brandt Stephen J. 
Published: October 2019

Abstract
The TAL1 (also known as SCL and TCL-5) protein is a class II basic helix-loop-helix (bHLH) transcription factor that plays an important role during embryonic and adult hematopoiesis. We previously established that the Protein 4.2 (P4.2) gene is a physiologic target of TAL1 in erythroid cells and showed that tandem E box-GATA elements in its proximal promoter mediated TAL1-directed gene activation. Through database searches, we identified two E box-GATA consensus sequences in the proximal promoter of another erythroid membrane skeleton gene, that encoding the Band 3 protein. Identical to what was observed for P4.2, overexpression of wild-type TAL1 slightly increased while enforced expression of a DNA binding-defective TAL1 mutant severely reduced endogenous Band 3 gene expression in murine erythroleukemia (MEL) cells induced to differentiate with dimethyl sulfoxide (DMSO). Overexpression of Ldb1 significantly inhibited and a LIM protein interaction-defective Ldb1 mutant virtually ablated Band 3 mRNA accumulation in DMSO-induced MEL cells. Quantitative chromatin immunoprecipitation (ChIP) analysis with Tal1 antibody confirmed that Tal1 was associated with the proximal promoter of the Band 3 gene in MEL cells, while analysis of approximately 28 kb of genomic sequence spanning the Band 3 gene, including 5 kb 5′ and 3′ of the gene, revealed another consensus E box-GATA element in intron 16; however, no significant TAL1 binding was detected in this region by ChIP analysis. To identify potential non-conventional TAL1 binding site(s), scanning ChIP analysis was applied to the entire 28 kb Band 3 genomic region, and a region ~2.5 kb upstream of the major transcriptional start site was found to bind significantly more TAL1 protein than the proximal promoter. Fine mapping of TAL1 binding to this region by ChIP analysis using more highly sheared DNA and smaller sized amplicons narrowed TAL1 binding to a region of ~100 bp, which we designated as the Band 3 upstream regulatory region (B3URE). Luciferase reporter assays in transiently transfected MEL cells with vectors containing genomic sequence revealed the presence of an orientation- and position- independent repressor in this upstream region, with a 107 bp fragment retaining nearly all the repressing activity of a larger, 700 bp fragment. In contrast, coexpression of TAL1, E47, GATA-1, LMO2, and Ldb1 in COS cells led to transactivation of a reporter gene linked to the promoter-proximal E box-GATA element. These data suggest that TAL1 acts as a repressor on the upstream B3URE in undifferentiated cells and then, as differentiation proceeds, as an activator on the E box-GATA elements in the proximal promoter.