bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytosolic Histone H1.2 Releasing under Different Apoptotic Stimuli in Chronic Lymphocytic Leukemia (CLL).
Authors: Gine Eva , Crespo Marta , Muntanola Ana , Montserrat Emili , Bosch Francesc 
Published: October 2019

Abstract
Recently, it has been shown that nuclear histone H1.2 is released into cytoplasm when apoptosis is induced by DNA double-strand breaks (DSB’s), this process being dependent on p53 functional status. In addition, cytosolic histone H1.2 induces cytochrome C release in a Bak-dependent manner. Thus, cytosolic histone H1.2 release represents a new mechanism that links DSB’s with activation of the apoptotic mitochondrial pathway. Against this background, we analyzed the release of histone H1.2 in the cytosol of purified CLL cells during apoptosis induced by fludarabine (F), mitoxantrone (M), etoposide, or ionizing radiation. In addition, the presence of histone H1 was correlated with p53 functional status. Cell viability and analysis of apoptosis were investigated by annexin V/PI staining and FACscan analysis. The presence of histone H1 and H1.2 in the cytosolic fraction was assessed by Western Blott using the anti-histone H1 (Upstate) and anti-histone H1.2 (Abcam) antibodies. Histone H1 traffic was also evaluated by using immunofluorescence analysis in CLL cells suspensions. FISH analysis was used to select samples with (n=3) or without (n=6) p53 deletion, and activation of p53 after treatment was assessed by Western Blot. In cases without p53 deletion, increased apoptosis was observed under all stimuli, the FM combination being the most effective. In such cases, histone H1.2 release was apparent 6 hours after the onset of irradiation or pharmacologic treatments, progressively increasing up to 24 hours. In contrast, cases with p53 deletion displayed a low cytotoxic effect upon different treatments. Interestingly, no p53 activation or histone H1.2 release into cytosol was observed. These results were also confirmed by immunofluorescence analysis, in which histone H1.2 was only visible in the cytosol of non-deleted p53 cases. These results demonstrate that, upon drug or irradiation exposure nuclear histone H1.2 is released into the cytoplasm of CLL cells in a p53-dependent manner. This suggests that, in CLL, histone H1.2 traffic contributes to the apoptosis induced by DSB’s and to drug resistance in cases with p53 deletion.
Randomized Comparison of Cladribine with Cyclophosphamide and Fludarabine with Cyclophosphamide in Previously Untreated Progressive and Symptomatic Chronic Lymphocytic Leukemia: The Interim Analysis of PALG CLL3 Trial.
Authors: Robak Tadeusz , Blonski Jerzy Z. , Gora-Tybor Joanna , Najda Jacek , Stella-Holowiecka Beata , Calbecka Malgorzata , Kowal Malgorzata , Dmoszynska Anna , Ceglarek Bernadeta , Konopka Lech , Szpila Tomasz , Warzocha Krzysztof , Piszcz Jaroslaw , Kloczko Janusz , Zawilska Krystyna , Mital Andrzej , Hellmann Andrzej , Potoczek Stanislaw , Kuliczkowski Kazimierz , Sulek Kazimierz , Zdunczyk Andrzej , Kasznicki Marek 
Published: October 2019

Abstract
The efficacy and toxicity of cladribine (2-CdA) + cyclophsphamide (CY) - CC versus fludarabine (FA) + CY - FC were compared in previously untreated chronic lymphocytic leukemia (CLL) patients. The study was started in January 2004. Eligible patients were asigned to either 2-CdA 0,12mg/kg/d and CY 250mg/m2/d for 3 consecutive days or FA 25mg/m2/d and CY 250mg/m2/d also for 3 days. Courses were repeated at 28 days intervals or longer if myelosupression and/or infection developes for a maximum 6 courses. The response and toxicity criteria were those recommended by NCI SWG. Minimal residual disease (MRD) was evaluated by flow cytometry if complete response (CR) was achieved. As shown in the table, there were no significant differences in the rates of overall response (OR), CR, grade 3/4 thrombocytopenia, neutropenia and infections between the programmes. MRD negativity was obtained in 4 patients in CC arm and in 6 patients in FC arm (p=0.69). The death rate was similar in both groups, 2 (3.2%) and 6 (8.2%), respectively (p=0.19). The therapy related mortality was not observed. In conclusion, CC and FC programmes seem to have similar efficacy and toxicity in previously untreated CLL patients. The results of the interim analysis of PALG CLL3 trial justify continuation of this study.
Treatment Pts enrolled Pts evaluated OR (%) CR (%) Thrombocytopenia Neutropenia Infections CC 96 63 57 (90.5) 20 (31.7) 8 (12.7) 15 (23.8) 26 (41.3) FC 100 73 62 (84.9) 25 (34.3) 6 (8.2) 17 (23.3) 20 (27.4) p value 0.24 0.45 0.27 0.53 0.06
Recombinant Mini-β3 Integrin Fragments for the Detection of HPA-1 Antibodies.
Authors: Stafford Prachi , Ghevaert Cedric , Campbell Kate , Smith Graham , Williamson Lorna , Watkins Nicholas A. , Ouwehand Willem H. 
Published: October 2019

Abstract
The αIIbβ3 integrin (GPIIbIIIa, CD61) is the platelet receptor for fibrinogen, fibronectin and vitronectin. A single nucleotide polymorphism (dbSNP Id= rs5918) in the ITGB3 gene defines the Human Platelet Antigen (HPA)-1 system encoding either a Leucine (1a) or Proline (1b) at position 33. HPA-1a immunisation is of clinical relevance in neonatal alloimmune thrombocytopenia (NAIT), post transfusion purpura (PTP) and platelet refractoriness. Currently, the detection of HPA-1 antibodies is based on a cumbersome sandwich ELISA requiring HPA genotyped platelets as source of antigen. We set out to express soluble calmodulin (CaM) tagged β3 fragments suitable for HPA-1 antibody detection. The design of fragments was informed by domain structural information, analysis of homology between β3 and integrin-like monomeric proteins and the evolutionary β3 domain phylogeny. Combined with computer modelling, this was the basis of the successful expression in Drosophila S2 cells of eight β3 fragments with either Leu33 or Pro33 (ΔSDL, ΔβA, PSI-Hybrid and PSI alone, both alleloforms for each of the four). Murine monoclonal antibodies Y2/51, SZ21 and CRC54 reacted with the β3 fragments as expected with only CRC54 reacting with the PSI fragment. Two of 3 human HPA-1a specific phage antibodies (19–7, 23–15, both derived from a PTP patient) reacted with all 4 Leu33 fragments but CamTran007 derived from a NAIT patient only reacted with the ΔSDL-Leu33 and ΔβA-Leu33 fragments, defining an epitope split. By ELISA, reactivity of both potency (NIBSC-03/152) and sensitivity (NIBSC-93/710) standards were comparable to those seen by MAIPA. Twelve polyclonal samples from NAIT patients (5 with cerebral haemorrhage [CH], 5 with platelet counts <40x109/L but no CH and 2 with counts >40x109/L) of potency ranging from 5–193 au/ml were detected by ELISA with Leu33 forms of ΔSDL and ΔβA. In conclusion soluble mini-β3-Leu33 fragments have been successfully expressed and shown to react with all polyclonal anti-HPA-1a samples tested from NAIT patients. The CaM tagged β3 fragments are now being further validated with a repository of 150 HPA-1a antibody samples from a NAIT case series with well defined treatment and clinical outcome histories to establish the relation between HPA-1a antibody potency, epitope reactivity and clinical phenotype.
Suppression of NK Cell-Mediated Bone Marrow Cell Rejection by CD4+CD25+ Regulatory T Cells: Linkage of Adaptive to Innate Responses.
Authors: Murphy William J. , Bareo Isabel , Hanash Alan M. , Welniak Lisbeth A. , Sun Kai , Blazar Bruce R. , Levy Robert B. 
Published: October 2019

Abstract
While a link between the innate to adaptive immune system has been established, studies demonstrating direct effects of T cells in regulating Natural Killer (NK) cell function have been lacking. Naturally occurring CD4+CD25+ regulatory T cells (Tregs) have been shown to potently inhibit adaptive responses by T cells. We therefore investigated whether Tregs could affect NK cell function in vivo. Using a bone marrow transplantation (BMT) model of hybrid resistance, in which parental (H2d) marrow grafts are rejected by the NK cells of the F1 recipients (H2bxd), we demonstrate that the in vivo removal of host Tregs significantly enhances NK-cell mediated BM rejection. This heightened rejection was mediated by the specific NK cell Ly-49+ subset previously demonstrated to reject the BMC in this donor/host pairing. The depletion of Tregs could also further increase rejection already enhanced by treating recipients with the NK cell activator, poly I:C. Although splenic NK cell numbers were not significantly altered, increased splenic NK in vitro cytotoxic activity was observed from the recovered cells. The regulatory role of Tregs was confirmed in adoptive transfer studies in which transferred CD4+CD25+ Tregs resulted in abrogation of NK cell-mediated hybrid resistance. Thus, Tregs can potently inhibit NK cell function in vivo and their depletion may have therapeutic ramifications with NK cell function in BMT and cancer therapy.
Managing Erythropoietin Usage in a Large Veterans Healthcare System.
Authors: Karri Sirisha , Carter Gary , Safar Mazin , Kyasa Mouhammed Jameel , Mehta Paulette 
Published: October 2019

Abstract
Erythropoietin (EPO) is used for many hematologic and other diseases and for adjunctive, supportive care of patients with cancer undergoing chemotherapy. However the cost is high, and excessive use may cause complications and/or compromise cancer survival. EPO utilization was the singlest largest pharmacy expense in the Central Arkansas Veterans Healthcare System and averaged $780,000 in fiscal year 2002–2003, with projections of over $2 million for the following year. We therefore studied methods to manage EPO usage through education, guideline development, monitoring, and through development of EPO-clinics modeled after coumadin-clinics. We designed and disseminated guidelines and algorithms for use of EPO, using national guidelines tailored to the specific needs of veterans and the Veterans Healthcare System. Guidelines for monitoring EPO included ongoing surveillance of iron status including iron and TIBC, ferritin, and reticulocyte hemoglobin measurements. Reticulocyte hemoglobin measurements were used as an immediate parameter of iron reserve and compared to iron profile as an indicator of iron status. Other monitoring included patient symptoms, blood counts, and course of disease. We stopped automatic refills of medication and required new prescriptions for each 6-week period so as to assure re-evaluation of the continued need for EPO and to assure titration of dosage to optimal hemoglobin and patient condition. We also flagged EPO orders in the V.A. computerized patient record system (CPRS) so that we could monitor compliance to guidelines. A pharmacy subcommittee reviewed patient profiles, symptoms, laboratory parameters, compliance to guidelines, and outcomes in relation to EPO usage. We also initiated an EPO-clinic, modeled after coumadin-clinics in which pharmacists, nurses, and physicians worked together to counsel patients about EPO, to titrate dose to best response, to prevent excess dosing, and to provide ongoing outpatient, inpatient and at-home education. Prior to initiation of the program, the projected annual cost of EPO was $2 million; after initiation of the program, the actual cost was $l.l5 million, or 43% less than the total projected amount. The program also resulted in greater attention to EPO usage, better staff and patient education, and to piloting of an EPO-clinic to coordinate medical, pharmacy, and psychosocial resources into a single clinic. This model may be effective in use of other, highly effective but very expensive medications being used in patients with hematologic disorders.
Effects of STI571 on the Expression of c-kit Receptor in Mononuclear Cells from the Bone Marrow of Patients with Acute Non-Lymphocytic Leukemia.
Authors: Feng Ru , Huang Fen , Tantai Linfang 
Published: October 2019

Abstract
Objective: To explore the effects of STI571 on the expression of c-kit gene of the bone marrow mononuclear cells from patients with acute non-lymphocytic leukemia (ANLL) and to study the gene difference expression in original leukemia cells from the bone marrow of ANLL patients with CD117 overexpress treated by STI571, so as to explore initially the mechanism of the restraint of proliferation caused by STI571.
Methods: The expression of CD117 and C-kit mRNA level were assayed by FACS and RT-PCR before and after different concentration STI571 cultivatio. The changes of gene expression were examined with the cDNA array in original leukemia cells after treated by STI571 and control.
Results: After STI571 treatment, the expression of CD117 and the mRNA expression of c-kit were significantly lower than before (P<0.05). There were over 730 different genes in each pair of chips, in which 156 different genes expressed together. In all different genes, 66.67% (104/156) were up-regulated, 33.33% (52/156) were down-regulated. Cellular skeleton accounted to 4.49% (7/156), oncogenes and tumor supressors 6.41% (10/156), cells receptor 7.05% (11/156), extracellular signal transduction proteins 23.07% (36/156), cyclin proteins7.05% (11/156), apoptosis correlation proteins 1.92% (3/156) and other genes accounted to50% (78/156). Genes with over ten times relative difference amounted to 26.92% (42/156).
Conclusion: The restraint of c-kit on ANLL fresh cells was related to concentration of STI571. The mechanism of the restraint of proliferation caused by STI571 may relate to three ways: 1. STI571 realizes its biological function by affecting different signal conduction pathways through protein tyrosine kinase, guanine-nucleotide-binding protein and protein kinase C; 2. STI571 restrains the infinite differentiation of cells through affecting of tumor cellular mitosis; 3. Destroy cellular skeleton.
Combination of PP1/PP2 and LY294002 Induce Apoptosis Efficiently to TEL-FGFR3 Transfected Ba/F3 Cells.
Authors: Okamura Daisuke , Yagasaki Fumiharu , Maeda Tomoya , Ishikawa Maho , Jinnai Itsurou , Bessho Masami 
Published: October 2019

Abstract
We previously reported that TEL-FGFR3 in a patient with peripheral T-cell lymphoma and AML conferred IL-3 independency to Ba/F3 cells and activates MAPKs, p38, PI3K, Stat3 and Stat5 through its constitutive tyrosine kinase (TK) activity in TEL-FGFR3 transfected Ba/F3 cells (TF-V5). Both FGFR3 TK specific inhibitor SU5402 and PI3K inhibitor LY294002 inhibited cell growth of TF-V5 in a dose dependent manner (IC50=5μM, 10μM respectively). SU5402(25μM) resulted in G1 arrest (90%) and induced 80% of apoptosis to TF-V5 after 24hr. LY294002(50μM) decreased expression of c-Myc, Cyclin D3 and CDK4 in TF-V5 and induced G1 arrest (90%) and apoptosis about 20% to TF-V5 after 24hr. SU5402 completely suppressed expression of Bcl-xL and Bcl-2 after 24hr. LY294002 partially suppressed these expressions. As LY294002 did not affect the phosphorylation of STAT5 and STAT3, we hypothesized that these distinct apoptosis inducing abilities between SU5402 and LY294002 might be caused by the difference of Bcl-xL and Bcl-2 expression levels. Recent report suggests that FGFR3 activates Stat5 through recruitment of Pyk2 (proline-rich tyrosine kinase 2)-Src to its juxta-membrane (JM) domain. Therefore, we examined whether PP1/PP2 which are known as Src kinase inhibitors inhibit TF-V5 cell growth. PP1 and PP2 inhibited TEL-FGFR3 induced cell proliferation in a dose dependent manner (IC50=15μM, 15μM respectively). In contrast, growth of mock with IL-3 was not inhibited at a high concentration of PP1or PP2(30μM). PP1/PP2 did not affect auto-phosphorylation of TEL-FGFR3. Interestingly, PP1/PP2 inhibit Tyr phosphorylation of Pyk2 except Tyr402 and markedly suppressed activation of Stat3, Stat5 without affecting MAPKs and PLC gamma activation. PP1/PP2 suppressed expression of Bcl-xL and Bcl-2 partially but not of Bax and induced apoptosis 20% and 30% respectively. PP1/PP2 partially blocks cell cycle at G1 phase. When TF-V5 was co-treated with PP1(30μM)+LY294002(50μM) or PP2(30μM)+LY294002(50μM), apoptosis was induced in 60–70% of TF-V5 cells which remained at G1 phase after 24hr. Although PP1+LY294002 or PP2+LY294002 did not suppress Bcl-xL and Bcl-2 completely like SU5402, PP1/PP2 induced N-terminal cleavage form of Bax.
Conclusion: PP1/PP2 induces apoptosis in TF-V5 by activating Bax and by Pyk2-Src kinase inactivation that leads to the change of Bax/Bcl-2 or Bax/Bcl-XL ratio. Combination of PP1/PP2 and LY294002 induce high rate of apoptosis under G1 arrest in TF-V5 and may represent a novel approach against TEL-FGFR3 associated hematopoietic malignancies.
Trisomy 11 and Partial Tandem Duplication of the MLL Gene in a Case of Infant Acute Myeloid Leukemia M0.
Authors: Serravalle Salvatore , Purgato Stefania , Melchionda Fraia , Tonelli Roberto , Pession Andrea 
Published: October 2019

Abstract
Trisomy 11 is uncommon in pediatric acute myeloid leukemia (AML) and molecular studies of AML with trisomy 11 in adult patients have recently revealed high incidence of partial tandem duplication (PTD) of the MLL gene. We report the first case of trisomy 11 and MLL-PTD in a 3 months old girl with AML, the immunophenotype of bone marrow aspirate documented blast cells positive for CD34, CD33, AntiHLA DR, CD117, and MPO <3%, M0 according to FAB classification. No exposure to potential mutagenic agents during pregnancy was documented. The most frequent translocations of MLL, t(4;11) and t(9;11), were investigated by RT-PCR at diagnosis and resulted negative, as well as other genetic abnormalities common in pediatric AML as inv 16, t(8;21), t(15;17), FLT-ITD, D835 mut. Otherwise, interphase FISH analysis, performed with a specific MLL probe, revealed a MLL triple signal (Fig. 1a). Moreover trisomy 11 was confirmed by FISH analysis with a centromeric probe for chromosome 11 (Fig. 1b). RT-PCR analysis revealed PTD of an internal portion of MLL and direct sequencing confirmed the in-frame fusion of exon 6 and exon 2 (Fig. 1c). The self-fused MLL has a duplicated N-terminal region that contains AT hook DNA-binding and DNA methyltransferase motifs. MLL-PTD may involve different exons, and it is known to be associated with a poor prognosis. Even if the oncogenic mechanism of MLL-PTD is not clear yet, several hypothesis suggested a role of DNA binding motifs duplication in the increase of affinity for MLL binding to a target site, or in the separation of autoregolatory domains from their sites of action. Beside its significance as a prognostic marker at diagnosis, MLL-PTD may be a target for PCR-based detection of minimal residual disease in AML patients.
Figure 1. Interphase FISH analysis with MLL probe revealed a MLL triple signal (a) and with a centromeric probe confirmed trisomy 11 (b). Direct sequencing confirmed the MLL PTD with in-frame fusion of exons 6 and exon 2(c). Figure 1. Interphase FISH analysis with MLL probe revealed a MLL triple signal (a) and with a centromeric probe confirmed trisomy 11 (b). Direct sequencing confirmed the MLL PTD with in-frame fusion of exons 6 and exon 2(c).
Different Patterns of Cytogenetic Evolution in CML Treated with Imatinib.
Authors: Haase Detlef , Steidl Christian , Bierbaum Anne , Soeling Ulrike , Truemper Lorenz , Griesinger Frank 
Published: October 2019

Abstract
In CML the Ph+ cell clone is genetically unstable leading to the acquisition of typical additional abnormalities during accelerated phase and blast crisis in up to 60 to 80 % of pts. Imatinib, by selectively inhibiting bcr-abl tyrosinkinase induces complete cytogenetic remissions in up to 80% of pts. Beneath others, two main mechanisms are responsible for the development of Imatinib-resistance: On the molecular level mutations of the bcr-abl domain and on the cytogenetic level acquisition of additional abnormalities can occur. To investigate the latter phenomenon a cohort of 67 pts treated with Imatinib was karyotyped sequentially. Treatment results were as follows: CCR: 40 (60%), MaCR: 4 (6%), MinCR: 9 (13.4%), NR: 7 (10.5%), cytogenetic progression: 5 (7.5%). Five different patterns of cytogenetic progression under Imatinib were observed. I (occurrence of an additional abnormality): One case with initial t(3;9;22) and additional 17p-/P53 allelic loss 36 after months under Imatinib. II (expansion of an initially present additional abnormality): Initial t(9;22) and a subclone with additional t(2;3), followed by expansion of the sublone with additional t(2;3). III (occurrence of multiple additional, partly atypical abnormalities and expansion of this clone): One case with initial t(9;22) and additional inv(3q), 7q-, +19, +Ph. IV (occurrence of typical additional abnormalities and expansion of the clone with the highest number of abnormalities): Initial t(9;22) and additional +8,+Ph. V (discordant response): Initial t(9:22), subsequent MaCR with parallel occurrence of a clone with additional +8,+8,+Ph and further progression. Of the five cases with cytogenetic progression two pts (I,V) had been treated with Imatinib only, one pt had a combination of Imatinib and IFN (II) and two pts (III, IV) had low-dose AraC before Imatinib. It can be concluded that pretreatment with AraC might have modulated the genetic response to Imatinib possibly leading to genetic evolution via selection of cytogenetically resistant clones in 2/5 cases. In 3/5 cases intrinsic mechanisms must have contributed to Imatinib-resistance. As demonstrated by case V our observations furthermore draw the attention to the underestimated problem of discordant genetic responses only traceable by classical cytogenetics. Thus the therapeutic consequences of these findings have not been clearly defined as yet.
Multiplex Fluorescence In Situ Hybridization (M-FISH) in the Detection of Complex Karyotypic Abnormalities of Acute Myeloid Leukemia and Myelodysplastic Syndromes.
Authors: Li Jianyong , Pan Jinlan , Xiao Bing , Ma Li , Qiu Hairong , Li Li , Xu Wei , Xue Yongquan , Ruan Changgeng 
Published: October 2019

Abstract
The complex chromosome abnormalities (CCAs) were one of the most important poor prognostic risk factors in acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). Chromosome analysis using classical cytogenetic banding techniques fails to completely resolve complex karyotypes and cryptic translocations. The technique of multiplex fluorescence in situ hybridization (M-FISH) allow for the simultaneous visualization of all chromosomes of a metaphase in a single hybridization step and thereby enable to comprehensively analyze complex karyotypes and the identification of new and cryptic translocations. To investigate the value of M-FISH in the detection of complex karyotypic abnormalities of AML and MDS. M-FISH was used in combination with interphase-FISH to study 24 cases of AML and MDS with CCAs showed by R-banding of conventional cytogenetics (CC). In 14 cases of AML with CCAs, 4 gains of whole chromosome and 4 losses of whole chromosome were confirmed by M-FISH, while 12 losses of whole chromosome were revised as derivative chromosomes resulted from various structural aberrations. 26 derivative chromosomes and 19 marker chromosomes were characterized precisely by M-FISH. Most of them were unbalanced translocations, including 2 complex t(8;21), which have not been previously described:t(8;21), der(8) t(8;21) (8pter→8q22::21q22→21qter), der(21) t(8;21;8) (8qter→ 8q22::21p13→ 21q22::8q22→ 8qter) and t(21;8;18;1), der(8) t(8;21) (8pter→ 8q22::21q22→ 21qter), der(21) t(21;8;18;1) (21p13→ 21q22::8q22→ 8q24::18?::1q?q?). In 10 cases of MDS, 37 kinds of structural rearrangements were detected by M-FISH including insertion, deletion, translocation and derivative chromosomes, and among them 34 kinds were unbalanced rearrangements, only 3 were balanced rearrangements including t(6;22)(q21;q12), t(9;19)(q13;p13) and t(3;5)( ?;?), 7 abnormalities were never reported before. The CCAs invloved nearly all chromosomes, of which the chromosome 17, 5 and 7 were invloved more frequent than the rest. Chromosomes 5, 17, 7 were involved in 15 cases (62.5%), 12 cases (50%) and 6 cases (25%) respecrively. We conclude that M-FISH could refine CCAs of AML and MDS patients, find or correct the missed or misidentified aberrations by CC analysis. Our findings confirm that M-FISH is a powerful tool to characterize complex karyotypes in AML and MDS.