bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tashinone IIA Can Trigger the Apoptosis of a Megakaryocytic Cell Line CHRF- 288 through the TNFRSF9 Mediated Caspase-3 Activation Pathway
Authors: Liu Chang Charlie , Wang LiangJie Vicky , Zhou Min , Ye JieYu , Chan Godfrey ChiFung , Yang Mo 
Published: April 2020

Abstract
Tanshinone IIA (TIIA), a purified compound and an active component of Salvia miltiorrhiza (DanShen) has previously been shown to induce apoptosis on several human cancer cell lines. In the current study, we studied the cytotoxic effect of TIIA on a megakaryocytic leukemic cell line CHRF-288. First, we tested the cytotoxic effect of TIIA on CHRF-288 cells by Trypan blue assay and the cell viability was reduced to 80.8% after 72hrs of TIIA treatment (30ug/ml). Using Annexin V/PI staining and flow cytometry, we confirmed that TIIA induced apoptosis on CHRF-288 cells in a dose dependent manner. At the concentrations of 1, 3, 10 and 30ug/ml, the early apoptosis cells proportion showed a stepwise increment of 11.8±1.2%, 13.2±2.8%, 16.3±0.8% and 22.4±1.5% at 72 hrs respectively. This was verified by Caspase 3 assay which showed the activation of Caspase 3 increased from 5.1% to 16.2% after 10ug/ml of TIIA treatment. Activation of Caspase 3 implied the involvement of common apoptotic pathway. Then, we studied the mechanisms of apoptosis. Using JC-1 assay, we found that the depolarized cells increased from 9.06% to 16.6% after 10ug/ml TIIA treatment suggestive the involvement of intrinsic apoptotic pathway. To further determine the molecular mechanisms involved in the pro-apoptosis effect, Microarray studies using Affymetrix 133 plus genechips were conducted to identify the genes that were differentially expressed after TIIA treatment. Several groups of genes involved in apoptosis, calcium regulation and cell cycle checkpoints were found to be differentially expressed after the treatment. The differential expressions of these genes were validated using quantitative PCR. The most significantly upregulated genes (5.9±0.333 folds) was TNF Receptor Super-Family 9 (TNFRSF9). In addition, Receptor Interacting Protein Kinase (RIPK), a protein that likely interacts with TNFRSF9, was upregulated to 1.7±0.167 folds. And lastly, Jun, a transcriptional factor of TNFRSF9 was up-regulated to 1.7±0.03 folds. Since TNF and RIPK1 both involve in caspase-8 activation, this was suggestive of the activation of external apoptotic pathway. Our findings suggested that TIIA could induce significant apoptosis on mekakaryoctic leukemic cells via both intrinsic and extrinsic apoptotic pathways. Its role in treating this poor prognostic leukemia deserves further exploration.
Role for Chemokines in the Vasculopathy Associated with Sickle Cell Dsease and Pulmonary Hypertension
Authors: Tailor Anitaben , Kato Gregory J. 
Published: April 2020

Abstract
Sickle cell disease (SCD) is a hemolytic disorder characterized by transient vaso-occlusive crisis (VOC). Moreover, there is a high prevalence of SCD-associated pulmonary arterial hypertension (PAH), indicated by a tricuspid regurgitant jet velocity (TRV) ≥ 2.5 m/sec, leading to increased risk for early mortality. Monocytes have been proposed as pivotal cells in vascular inflammation in animal models of SCD. Preliminary studies in our group have shown a potential role for monocyte chemokines in the vasculopathy of SCD. The objective of this study is to investigate the role of two chemokines: regulated upon activation, normal T cell expressed and secreted (RANTES) and macrophage inflammatory protein-1β (MIP-1β) in SCD and SCD-associated PAH. Plasma samples were collected from patients with SCD at steady state and from healthy African-American control subjects. Plasma levels of RANTES and MIP-1β were measured using an enzyme immunoassay system. Patients with SCD exhibited significantly higher levels of both MIP-1β (median 465.3 vs. 398.5 pg/ml, p&lt;0.0001) and RANTES (median 47774 vs. 17445pg/ml, p&lt;0.0004) than healthy controls. Further characterization of SCD patients with or without PAH demonstrated that whereas MIP-1β exhibited no correlation with PAH, RANTES demonstrated an inverse correlation with PAH (r= −0.25, p&lt;0.03) with significance achieved in patients that exhibited severe PAH (TRV&gt; 2.9, p&lt;0.05). These findings suggest that
both chemokines, RANTES and MIP-1β, are associated with the vasculopathy of SCD, and whereas MIP-1β does not appear to be associated with SCD-associated PAH, RANTES levels are inversely correlated to the severity of PAH. RANTES merits further investigation as a potential marker in PAH, and for a possible role in its progression.
INK4 Locus Deletion in Childhood Acute Lymphoblastic Leukemia (ChALL)-a Preliminary Clinical Evaluation
Authors: Paisiou Anna , Papadhimitriou Stefanos I , Tsagarakis Nikolaos , Papadakis Vassilios , Kentrou Nektaria , Paterakis Georgios , Parcharidou Agapi , Androutsos Georgios , Tsitsikas Konstantinos , Kostopoulos Ioannis , Pavlidis Dimitris , Georgaki Maria , Polychronopoulou Sophia 
Published: April 2020

Abstract
Introduction: Deletions of the INK4 locus (on chromosomal region 9p21) are common in chALL. p16/14 and p15 genes lost with INK4 deletion are tumor suppressors, with a well-documented role in regulating cell cycle and chemosensitivity and in promoting apoptosis in vitro. However, the clinical significance of INK4 deletions remains unclear.
Patients & Methods: The study included 78 children (49 boys and 29 girls) with ALL (medium age: 3,12, range:0,7 to 16,5 ). The disease was of B- and T-cell origin in 70 and 8 cases, respectively. INK4 deletion was investigated by interphase FISH on the diagnostic bone marrow samples and the result was correlated with the patients’ demographic data, biological and clinical parameters, and disease outcome. All patients were treated according to the ALL-BFM-95 protocol and followed-up for 11–105 (medium 41) months.
Results: INK4 locus deletion was found in 24 cases (30.8%). The loss was hemizygous in 15 (19.2%) and homozygous in 9 cases (11.5%). A positive finding was strongly associated with disease of T-cell origin (p=0.008). Apart from a weak correlation between INK4 deletion and advanced age (p=0.068), there was no association with sex, initial leucocytosis level, cytogenetic category, risk group grade according to the ALL-BFM- 95 criteria or the presence of residual disease at days 15 and 33. Nevertheless, INK4 homozygous deletion was significantly associated with disease-free (p=0.004) and overall survival (p=0.002), while the respective associations for an hemizygous deletions were found insignificant (p=0.137 and p=0.131).
Conclusions: The findings from this cohort of children suggest that INK4 homozygous deletion is an unfavorable prognostic factor in chALL, even for cases in which an aggressive course cannot be predicted by the conventional risk estimating criteria. Unclear results with regard to hemizygous deletion may be attributed to the functional state of remaining p16/p14 and p15 genes on the non-deleted chromosome 9. The study of a greater group of patients and the extension of the follow-up period will hopefully clarify whether INK4 deletion is an independent prognostic factor and whether it could be incorporated in risk evaluation guidelines, especially for cases which would otherwise be treated as chALL of standard or medium risk.
Combination of Bortezomib, Dexamethasone, Thalidomide and Zoledronic Acid (VdtZ) Results in a High Rate of Complete Response in High-Risk Non-Hodgkin Lymphoma
Authors: Teoh Gerrard , Ang Ai Leen 
Published: April 2020

Abstract
We previously reported that the combination of low-dose dexamethasone (d) and thalidomide (t) with higher frequency zoledronic acid (ZOMETA, Z) – the “dtZ” regimen – was associated with an overall response rate (RR) of 75.0% in patients with relapsed/refractory multiple myeloma (MM). However, the rate immunofixation (IF)-negative complete reponses (CR) was only 7.7%. In those patients who did not achieve CR, addition of bortezomib (VELCADE, V) to “dtZ” – the “VdtZ” regimen – greatly improved the overall RR to 92.9% and CR to 42.9%. In addition, 21.4% of “dtZ/VdtZ”-treated patients also achieved near-CR (nCR). Amongst 14 consecutive newly-diagnosed patients with MM who were treated with “dtZ/VdtZ”, the RR was 100% and CR/nCR was &gt;75%. Encouraged by these data in MM, we applied the 3-weekly “VdtZ” regimen – intravenous Vel 1.5 mg/m2 days 1, 4, 8 and 11; oral Dex 20 mg days 1, 8 and 15; oral Thal 100 mg days 1–21; and intravenous Zol 4 mg day 1 – in 3 consecutive patients with newly-diagnosed high-risk, difficult-to-treat non-Hodgkin lymphoma (NHL) to determine the feasibility of using “VdtZ” to treat NHL. The patient characteristics and results are summarized in the table below:
In summary, all 3 (100%) patients rapidly reported CR upon initiation of “VdtZ. Although significant tumor lysis was reported in all patients, treatment was fairly welltolerated. These data provide a basis for further studies on the “VdtZ” regimen for the treatment of high-risk NHL.
Parameter Case 1 Case 2 Case 3 PTCL = peripheral T cell lymphoma; DLBCL = diffuse large B cell lymphoma Age, Sex 78 years, Male 61 years, Female 61 years, Male NHL PTCL DLBCL DLBCL Stage IV B E S IV B E S III B E S Tumor burden Very bulky Extremely bulky Bulky Additional features Pleural effusion, ascites Massive ascites, pleural effusion Severe autoimmune hemolytic anemia Zubrod score 4 4 2 IPI risk High High High Response to“VdtZ” CR after 2 cycles CR after 3 cycles CR after 1 cycle Survival to date 12 months 4 months 1 month
Murine Hematopoietic Stem Cells Require Akt1 and Akt2 for Long-Term Self-Renewal
Authors: Juntilla Marisa M. , Patil Vineet , Joshi Rohan , Koretzky Gary A. 
Published: April 2020

Abstract
Murine hematopoietic stem cells (HSCs) rely on components of the Akt signaling pathway, such as FOXO family members and PTEN, for efficient self-renewal and continued survival. However, it is unknown whether Akt is also required for murine HSC function. We hypothesized that Akt would be required for HSC self-renewal, and that the absence of Akt would lead to hematopoietic failure resulting in developmental defects in multiple lineages. To address the effect of Akt loss in HSCs we used competitive and noncompetitive murine fetal liver-bone marrow chimeras. In short-term assays, Akt1−/−Akt2−/− fetal liver cells reconstituted the LSK compartment of an irradiated host as well or better than wildtype cells, although failed to generate wildtype levels of more differentiated cells in multiple lineages. When placed in a competitive environment, Akt1−/−Akt2−/− HSCs were outcompeted by wildtype HSCs in serial bone marrow transplant assays, indicating a requirement for Akt1 and Akt2 in the maintainance of long-term hematopoietic stem cells. Akt1−/−Akt2−/− LSKs tend to remain in the G0 phase of the cell cycle compared to wildtype LSKs, suggesting the failure in serial transplant assays may be due to increased quiesence in the absence of Akt1 and Akt2. Additionally, the intracellular content of reactive oxygen species (ROS) in HSCs is dependent on Akt signaling because Akt1−/−Akt2−/− HSCs have decreased ROS levels. Furthermore, pharmacologic augmentation of ROS in the absence of Akt1 and Akt2 results in an exit from quiescence and rescue of differentiation both in vivo and in vitro. Together, these data implicate Akt1 and Akt2 as critical regulators of long-term HSC function and suggest that defective ROS homeostasis may contribute to failed hematopoiesis.
Disulfiram, An Old Drug with New Potential Therapeutic Uses for Human Haematological Malignancies
Authors: Conticello Concetta , Martinetti Daniela , Lombardo Laura , Buccheri Simona , Adamo Luana , Amato Gabriella , Cavalli Maide , Chiarenza Annalisa , de Maria Ruggero , Giustolisi Rosario , Gulisano Massimo , Di Raimondo Francesco 
Published: April 2020

Abstract
Proteasome is a novel, interesting target in cancer drug therapy, and the proteasome inhibitor Bortezomib has been used for its antitumor activity in multiple myeloma and other lymphoid malignancies. The commonly used thiocarbamate alcohol-abuse deterrent disulfiram (DSF) is an aldehyde dehydrogenase (ALDH) inhibitor with documented proteasome-inhibiting activity. Recent reports indicate that DSF and other dithiocarbamates may have a significant potential in the treatment of human cancer without significant side effects. It was previously shown that this compound is able to block the P-glycoprotein extrusion pump, to inhibit the transcription factor nuclear factor-kappaB (NF-kB), to sensitize tumors to chemotherapy, to reduce angiogenesis, and to inhibit tumor growth in mice. In addition a recent work has described several compounds with proteasome-inhibiting activity, among which in the hit was DSF. Here we show that MM (18 samples), AML (14 samples) and ALL (5 samples) primary cells from newly diagnosed and relapsed/resistant patients were significantly sensitive to DSF at doses &lt; 5 mM (dose achievable in vivo). Median IC50 was DSF 0,5 mM for MM and DSF 0,35 mM for AML and r ALL samples after 48 hours of incubation. These cells are ALDH weakly positive and we did not find any difference in the sensitivity to DFS on the basis of amount of ALDH expression or status of disease (diagnosis vs relapse). Conversely, DSF, at the dose of 0,5 mM had only a weak effect on normal CD34 and peripheral blood mononuclear cells (&lt;30% of cell death). We next exposed the cells to the combination of low dose DCF (0,5 mM) plus CuSO4 (another commonly used herbal medicinal product, that makes complex with DCF and reduces the superoxide dismutase, an enzyme involved in the mitochondrial depolarization) and we found that this combination was able to induce about 80–90% of cell death after 48 hours of treatment. The apoptotic effect of the combination (DSF plus CuSO4 0,5 mM) was comparable to that exert by therapy with Bortezomib plus Desametazone for MM samples, Cytarabine, Etoposide, Daunorubicine alone or in combination for AML samples or Vincristine, Desametazone and Daunorubicine alone or in combination for ALL samples. All these drugs were used in vitro at doses compatible with the levels reached in vivo during cancer treatment. In addition we show that DSF plus CuSO4 induce loss of mithocondrial membrane potential thus suggesting the involvement of mitochondrial apoptotic pathway. These results may suggest a novel strategy for treating multiple myeloma and acute leukemias by employing an old drug, with known and mild side-effect, toward a new therapeutic use, probably in combination with other agents.
NTAL Potentiates Glucocorticosteroid-Induced Apoptosis in T-ALL – in Vivo Observations and Experimental Model
Authors: Svojgr Karel , Burjanivova Tatiana , Vaskova Martina , Kalina Tomas , Brdicka Tomas , Stary Jan , Trka Jan , Zuna Jan 
Published: April 2020

Abstract
PAG, LAT, NTAL and LIME belong to category of transmembrane adaptor proteins (TRAPs). They do not possess an enzymatic or kinase function, but they are involved in mediation of signal transmission from surface receptors to cell nucleus. We propose that some of them are engaged in development or maintenance of leukaemia.
We have previously demonstrated that expression status of TRAPs at mRNA level is specific in some subgroups of childhood ALL, particularly in B-cell precursor acute lymphoblastic leukaemia (BCP-ALL) with the TEL/AML1 fusion gene. Furthermore, we have described that variable expression of NTAL mRNA is related to the response to initial glucocorticosteroid pre-phase in the treatment of childhood T-ALL; patients with high NTAL levels show better treatment response compared to the low-NTAL cases.
In the current study, we aimed to prove experimentally that different levels of NTAL protein influence response of leukaemic T cells to glucocorticosteroids.
In the wild type Jurkat cells (human T-cell leukaemia cell line) the NTAL protein is undetectable. For the in-vitro experiments we created a derivative Jurkat cell line transfected with the NTAL construct. In the derivative cell line, the NTAL positivity at both mRNA and protein level was verified using RT-PCR and Western blotting. The derivative cell line in a cell culture behaves similar as wild type Jurkat cell line. Transfectants and wild type Jurkat cells were incubated with methylprednisolone, dexamethasone or with solvent alone (H2O) as a negative control. Using flow cytometry we determined a percentage of surviving cells after 24, 48 and 72 hours of treatment. Cells were stained with Annexin V and DAPI and living cells were defined as Annexin V, DAPI negative. At each time point, the number of living cells in the negative control was set to 100%. After 48 hours of dexamethasone treatment the number of surviving cells in the Jurkat wild-type was higher by 12% compared to the Jurkat cells expressing NTAL. After 72 hours this difference was even more prominent reaching 46%. The same effect of methylprednisolone treatment was less pronounced (non-significant difference at 48 hours and 5% difference at 72 hours). Based on our experimental data we propose that NTAL acts in T-cells as a putative tumour suppressor. A plausible mechanism of its function is that NTAL competitively inhibits another adaptor LAT, which is required for signal propagation to the nucleus in T-cells. NTAL and LAT can compete for its localisation in proper “lipid rafts” or for palmitoylation. Our data can be used for subsequent functional analysis of signaling pathways in leukaemic blasts as well as in physiological lymphoid cells.
Comparison of the Effects on Reversal of Multi-Drug Resistance of 5-Bromotetrandrine and Tetrandrine in K562/A02 Cell Line in Vivo and in Vitro
Authors: Chen Bao-An , Wang Jue-qiong , Cheng Jian , Gao Feng , Xu Wen-lin , Shen Hui-lin , Ding Jia-hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Sun Xin-chen , Cheng Hong-yan , Deng Yu-xia , Li Guo-hong , Liu Li-jie , Chen Wen-ji , Chen Ning-na , Wang Xue-mei 
Published: April 2020

Abstract
Objective This study was to compare the reversal effect of 5-bromotetrandrine (BrTet) with Tetrandrine (Tet) when combined with ADM on multidrug resistance cell line K562/A02 and to investigate the reversal mechanism of this new derivative.
Methods The protein levels of P-glycoprotein (P-gp) were detected by fluorospectrophotometry and Western blot. The mRNA levels of P-gp were determined by RT-PCR. The in vivo effect of Tet was investigated using nude mice grafted with sensitive human leukemia cell line K562 and MDR cell line K562/A02.
Results Flow cytometry assay showed that 1.0 μMol/L BrTet significantly increased the apoptosis percentage. BrTet also enhanced the intracellular accumulation of ADM in K562/A02 cells and its potency was greater than that of Tet at the same concentrations. BrTet inhibited the overexpression of P-gp and down regulated MDR1 mRNA expression in K562/A02 cells in a dose-dependent manner. In nude mice bearing K562 xenografts on the left flank and K562/A02 xenografts on the right flank, i.p. injection of 10 mg/kg BrTet significantly enhanced the antitumor activity of ADM against K562/A02 xenografts with inhibitory rates of 26.1%, while ADM alone inhibited the growth of KBv200 xenografts by only 5.8%.
Conclusion BrTet showed significant MDR reversal activity in vitro and in vivo. Its activity may be related to the inhibition of P-gp overexpression and the increase in intracellular accumulation of anticancer drugs, which lead to more K562/A02 cells apoptosis.
Megakaryocytic and Platelet Dysplasia in MDS Patients as Detected by Multiparameter Flow Cytometry Immunophenotyping
Authors: Sandes Alex Freire , Yamamoto Mihoko , Quijano Sandra , López Antonio , Chauffaille Maria de Lourdes L.F. , Oguro Tsutomu , Matarraz Sergio , Silva Marçal C.A. , Kimura Eliza Y.S. , Orfao Alberto 
Published: April 2020

Abstract
Introduction: Multiparameter flow cytometry immunophenotyping (FC) has shown the presence of phenotypic abnormalities in both CD34 precursors and maturing myeloid and erythorid cells in patients with myelodysplastic syndromes (MDS). However, detection of megakaryocytic dysplasia by FC still remains a challenge.
Aims: To evaluate the potential utility of FC analysis of megakaryopoesis and platelet immunophenotype in MDS patients for the diagnosis and prognostic evaluation of the disease.
Methods: A total of 54 peripheral blood (PB) samples corresponding to 39 patients with MDS and 15 controls (6 healthy subjects, 3 reactive cytopenias, 3 patients with May-Heglin syndrome and 3 myeloproliferative disorders) were collected in sodium citrate. In all cases, the median amount of expression of CD31, CD34, CD36, CD41a, CD41b, CD42a, CD42b, CD61, CD62P, CD63, PAC-1 and Fibrinogen receptor was evaluated on circulating platelets as refleted by the median fluorescence intensity (MFI). Morphologic megakaryocytic dysplasia was evaluated in parallel in bone marrow samples from the same MDS individuals.
Results: Morphological megakaryocytic dysplasia was detected in 49% of MDS patients who showed a shorter median overall survival (50 ± 8 vs 107 ± 12 months; p = 0.04). The following FC abnormalities were detected in circulating PB platelets of MDS patients: increased FSC and SSC (36%), overexpression of CD36 (18%), CD31 (36%), CD41b (20%), CD42b (2.5%) and CD61 (20%) and underexpression of CD36 (10%) and CD61 (5%). One case of aberrant CD34 expression was detected. A total of 27 (69.2%) patients presented at least one of these abnormalities by FC. Eighty percent of patients with morphological dysplasia and 64% without showed immunophenotypic abnormalities. According to IPSS, 6 patients constituted a high risk group (HRG) (Int-2 and High) and 28 a low risk group (LRG) (Int-1 and Low). Platelet phenotypic aberrations were detected in 83% of the HRG and in 75% of the LRG. Immunophenotypic abnormalities were not detected in myeloproliferative disorders. All May-Heglin cases and 2 reactive cytopenias presented overexpression of at least one of the following: FSC, SSC, CD31, CD36, CD41a, CD41b, CD42b and CD61. Deficient expression of CD36, CD61 and aberrant expression of CD34 were unique to MDS.
Conclusion: Our results show the presence of immunophenotypic abnormalities in PB platelets from MDS patients, the abnormally decreased reactivity for CD36 and CD61 together with aberrant CD34 expression being of potential diagnostic utility in MDS.