bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inactivation of JAK/STAT Cell Signaling by SK-7041, a Novel HDAC Inhibitor, Effectively Inhibits Growth of Acute Myeloid Leukemia Cells
Authors: Kim Byung-Su , Lee Chansu , Park Juwon , Ahn Kwang-Sung , Kim Byoung Kook , Park Seonyang , Lee Young-Yiul , Yoon Sung-Soo 
Published: April 2020

Abstract
Activation of the JAK/STAT pathway appears common in AML, occurring in up to 70% of AML patients. Therefore, JAK/STAT signal inhibitors are promising as candidate anti-cancer agents in AML. Recently, we reported that SK-7041, an HDAC inhibitor, inhibited the growth of AML cells via activation of caspase-3 and down-regulation of cyclin D1. These findings lead us to further examine whether SK-7041 inhibits the growth of KG1 AML cells via inactivation of JAK/STAT signals. Multi-immunoblotting technique (Kinetworks™ analysis) showed that expression of p-STAT-3, p-STAT-5, and p-Erk was down-regulated in KG1 cells treated with SK-7041. These results were confirmed by individual western blot analysis. In addition, IL-6-induced activation of STAT-3 and Erk was inhibited by treatment of SK-7041. Combined treatment of SK-7041 and JAK inhibitor (AG490) showed additive anti-leukemic effect as evidenced by caspase-3 activation, down-regulation of cyclin D1 (cMYC), and inhibition of phosphorylation of STATs (−1, −3). These results suggest that HDAC inhibitor, SK-7041, inhibited AML cell growth via inactivation of JAK/STATs pathway.
Pemetrexed Treatment of Breast Relapse in AML
Authors: Wang Chunyan , Tan Huo , Peng SiDa , Huang ZhenQian 
Published: April 2020

Abstract
Extramedullary relapse is a pattern of acute myeloid leukemia (AML) relapse post-allogeneic hematopoietic stem cell transplantation (alloHSCT). We present the case of a 43-year-old female patient with acute mix cell leukemia who had isolated extramedullary relapse develop in both breasts 2 years after alloHSCT, the first reported case in adult patients in the literature to the best of our knowledge. She received further chemotherapy with pemetrexed (Alimta) intravenous injection (500mg/D1, every 3 weeks is one course of treatment). After 2 courses of treatments, her swelled breasts were shrunk remarkably and no obvious side effects were found. We therefore conclude that pemetrexed treatment may be an effective regimen for breast extramedullary leukemia. Review of the literature about extramedullary relapse and breast involvement as well as the treatment of these cases were further discussed.
Clofarabine/Cyclophosphamide/Etoposide Combination Chemotherapy Well Tolerated in An Infant with Refractory AML
Authors: Rahman Shahera , Connor Philip P , Moppett John , Jenney Meriel , Cummins Michelle , Cornish Jackie 
Published: April 2020

Abstract
Refractory AML in children under one year of age is rare. Generally the high induction remission rate is offset by higher treatment-related mortality. We report a male infant who presented with generalised lymphadenopathy and hepatosplenomegaly aged 3 months. Morphology was consistent with M5 AML and cytogenetics showed a t(9;11)(p22;q23) MLL rearrangement. He was started on standard AML induction treatment for children under 1 year of age according to the AML15 protocol (daunorubicin 37.5mg/m2 days 1, 3, 5 with etoposide 75mg/m2 days 1 through 5 and cytarabine 75mg/m2 twice daily for 10 days). He never demonstrated count recovery and by day 25 post treatment, lymphadenopathy had returned. A bone marrow aspirate showed M5 morphology with 80% of cells showing MLL rearrangement by FISH. He was therefore taken off protocol and administered fludarabine 30mg/m2 and cytosine 2g/m2 once daily for 5 days and gemtuzamab ozogamicin 3g/m2 on day 1 (FLA-GO). At day 21 post chemo, his platelet count rose into the normal range but he remained neutropenic. A bone marrow showed some normal haematopoiesis but 33% of cells were blasts with MLL rearrangement by FISH. It was decided to repeat the FLA-GO but by day 25 post chemotherapy lymphadenopathy and organomegaly had returned and marrow examination showed 50% blasts with MLL rearrangement. At this point it was decided to administer clofarabine 25mg/m2 on days 1 to 4 with etoposide 75mg/m2 and cyclophosphamide 340mg/m2 on days 1 to 5. He tolerated the regimen well, the only side effect being myelosupression. By day 24 he had recovered peripheral counts, had no lymphadenopathy or organomegaly and a marrow showing 9% blasts with MLL rearrangement. He was able to proceed with allogeneic stem cell transplantation. This shows that clofarabine-containing combination regimens can be well tolerated in the very young.
Nature Products Enhance NKG2D Ligands Expression of CD123+CD34+CD38− Leukemia Stem Cells for Stimulating Cytotoxicity of NKG2D+ Cells to Themselves
Authors: Kunyuan Guo 
Published: April 2020

Abstract
CD123+CD34+CD38− leukemia cells regarded as leukemia stem cells, not only refractory to chemotherapeutics but also resistant to immune response such as cytotoxicity of prime natural killer cells, usually lead leukemia to relapse. It is essential to eliminate leukemia stem cells at utmost for cure. Here we would observe whether nature products could enhance CD123+CD34+CD38− leukemia cells to express NKG2D ligands for arousing cytotoxicity of NKG2D+ cells to thoroughly eliminate leukemia stem cells. First, CD123+CD34+CD38− leukemia cells, isolated from human acute leukemia cell line KG1a, whose MLL gene was broken apart and rearranged, of which over 60% were in G0 phase of cell cycle, which could form about 80 colonies per 1000 cells in methylcellulose after 21 days. What’s more, they were worse than CD123+CD34+CD38+ leukemia cells on resistance to cytotoxicity of natural killer cells(Killed ratio &lt;19%). CD123+CD34+CD38− leukemia cells from KG1a had characteristic of defined leukemia stem cells. Then, NKG2D+ cells from human peripheral blood monocytes were stimulated by cytokines and CpG, whose cytotoxicity against K562 cells reached (96.5±1.23)% analyzed by LDH releasing assay, suggesting NKG2D+ cells had better cytotoxicity than prime natural killer cells (69.31±8.45)%. However, cytotoxicity of NKG2D+ cells against CD123+CD34+CD38− leukemia stem cells was only(49.17±3.90)%, still higher than of prime natural killer cells (19±2.63)%. Therefore, whether to arouse cytotoxicity of NKG2D+ cells to deplete leukemia stem cells become essential. Finally, three kinds of nature products would be selected, including resveratrol (RV, a polyphenol found in berries), matrine and arsenic trioxide (ATO), which could decrease proliferation and survival of CD123+CD34+CD38− leukemia cells and whose 50% effective concentration (EC50) were respectively 56μM, 500μg/ml and 4.0μM. Importantly, EC50 of resveratrol could up regulate NKG2D ligands ULBP1 to (53.28±8.60)%, ULBP2 to (36.54±11.45)% and ULBP3 to (65.08±14.44)% on the surface of CD123+CD34+CD38− leukemia cells, and stimulate cytolytic ability of NKG2D+ cells, so that cytotoxicity of NKG2D+ cells against them reached from (49.17±3.90)% to (92.33±2.63)%. Similarly to matrine and arsenic trioxide, they could each up regulate ULBP2 from to 9.71±2.37% and ULBP1 to 24.38±3.65%, resulting in cytotoxicity of NKG2D+ cells against them reaching to 73.20±3.41% and 55.70±0.53%. In conclusions, cytotoxicity of NKG2D+ cells was better than of prime natural killer cells against not only K562, but also CD123+CD34+CD38− leukemia cells, which had characteristic of leukemia stem cells, even to resist killing of prime natural killer cells. Nature products, resveratrol, matrine and arsenic trioxide, could enhance NKG2D ligands expression of CD123+CD34+CD38− leukemia stem cells for stimulating cytotoxicity of NKG2D+ cells to leukemia stem cells. This finding could provide a new strategy for eliminate leukemia stem cells.
Rapid Genotyping of SNPs Influencing Warfarin Drug Response by SELDI-TOF Mass Spectrometry
Authors: Yang Shangbin , Xu Lihui , Wu Haifeng 
Published: April 2020

Abstract
The anticoagulant drug Warfarin exhibits significant inter-individual variability in dosing requirements. Different responses to Warfarin therapy are partly attributed to the single nucleotide polymorphisms (SNPs) that influence either Warfarin drug action (VKORC1) or drug metabolism (CYP2C9). Rapid genotyping of these SNPs is essential for clinicians to choose appropriate initial doses in order to quickly achieve anticoagulation effects and to prevent the complications associated with Warfarin overdoses. In this study, we explore the utility of surface-enhanced laser desorption and ionization time-of- flight (SELDI-TOF) mass spectrometry in the rapid genotyping of SNPs that control Warfarin drug sensitivity. The DNA containing the targeted SNPs is first amplified by polymerase chain reactions and then underwent the single base extension to generate specific SNP product. Afterwards, genetic variants displaying different masses are bound to Q10 anionic proteinChips and genotyped using a SELDI-TOF mass spectrometer in a multiplexed fashion. SELDI-TOF mass spectrometer offers a unique property of on-chip sample enrichment and clean-ups. Therefore, this genotype method eliminated many tedious experimental steps, such as sample desalting and concentrating, that are required prior to detection by standard mass spectrometers. The turnaround time for genotyping three known Warfarin sensitivity SNPs, CYP2C9*2, CYP2C9*3, VKORC1, is less than five hours. The analytical accuracy of genotyping detected by SELDI mass spectrometer has been confirmed by DNA sequencing. In summary, we have devised a novel multiplex genotyping method using a SELDI-TOF mass spectrometer. This test is fast, accurate, and therefore provides a superb clinical laboratory platform to promote the personalized health care in Warfarin therapy.
Practical Application of the 10 Mg Warfarin Initiation Nomogram
Authors: Wells Philip , Le Gal Gregoire , Tierney Sarah , Carrier Marc 
Published: April 2020

Abstract
Introduction: Initiation of warfarin therapy is a clinical challenge. A 10 mg warfarin initiation nomogram was recently validated in a randomized controlled trial. We sought to determine the efficacy and safety of this 10 mg warfarin initiation nomogram in “real-life” daily practice.
Methods: A retrospective cohort including all outpatients beginning concurrent treatment with warfarin and low-molecular-weight-heparin (LMWH) over a 24 month period in our Thrombosis Unit was reviewed.
Results: 841 patients were included, of them 640 (76.1%) were started on the nomogram. The nomogram was entirely followed in 324 patients (38.5%). The efficacy and safety profile was similar to that observed in the original clinical trial: 86% of patients managed according to the nomogram reached the INR target of 2.0 to 3.0 within 5 days. Mean duration of LMWH treatment was 6.0 ± 1.9 days, and 3.7% of patients had an INR ≥ 5.0 in the first four weeks of treatment.
Conclusion: The 10-mg nomogram effectively results into an early therapeutic INR with a good safety profile in “real life” daily practice.
Oncological and Hematological Neoplasms: Concomitant Oral Anticoagulation Therapy
Authors: Rupoli Serena , Lio Lidia Da , Scortechini Anna Rita , Ravaglia Francesca , Pulini Stefano , Mulattieri Simonetta , Picardi Paola , Canafoglia Lucia , Stronati Andrea , Tomassetti Simona , Tocchini Massimo , Leoni Pietro 
Published: April 2020

Abstract
The oral anticoagulant therapy (OAT) in oncologic patient is often problematic because both the haemorragic risk and the thrombotic risk, i.e. the recurrence of venous thromboembolism, are increased. Many studies have been conducted in patient with solid neoplasms; it remains still unclear whether these results could be entirely extrapolated to patients with haematological malignancies. Our aim was to analyse the clinical outcomes and the control of anticoagulation, measured as % time in target INR range, in the patients with cancer compared to patients without cancer; moreover we focused on patients with haematological neoplasms. A total of 500 patients were recruited in 6 years, regardless of the indication for OAT; 146 (29%) of them were affected by cancer (47 haematological and 99 solid neoplasms); those being treated with warfarin for less than 1 month were excluded from this analysis. It resulted that patients affected with cancer had a worse anticoagulation control, compared to those without cancer (55% vs 60%; p&lt;0.001). The rate of thrombosis was similar (6/146=4.1% vs 14/354=3.9%; OR 1.1); moreover the former had a tendency toward a higher rate of major bleeding complications (3/146=2.1% and 5/354=1.4% respectively; OR 1.5). As regarding hematologic patients we noted that the quality of anticoagulation control did not differ compared with solid malignancies (54% vs 55%, p = ns); no significant differences were found in the major bleeding (1/47=2.1% and 2/99=2.0% respectively; OR 1.05) and thrombotic complications (2/47=4.3% and 4/99=4.1% respectively; OR 1.05). The cancer patients discontinued warfarin primarily for surgical, endoscopic diagnostic or therapeutic procedures (32% vs 19%; p&lt;0.001); in particular the surgical procedures more frequently were the reason for discontinuation (27% vs 4% of the haematological patients, p&lt;0.001). The 47 haematological patients showed a younger median age compared to those with solid neoplasms (67 vs 72 years, p&lt;0.001), and discontinued OAT often because of invasive procedures (45% vs 26% of cancer patients, p&lt;0.02). In this experience the haematological patients seem not to have an increased risk of thrombosis recurrence and bleeding during OAT. Such a result probably reflects a good management of OAT even in presence of risk factors like central vein catheter or thrombocytopenia. Moreover, an appropriate initial decision of employing low-molecular-weight heparins (LMWH) rather than OAT in particular subsets of haematological patients (for example those with life-expectancy less than 2–3 months or undergoing high-dose chemotherapy for stem cell transplantation) can reduce the hemorragic risk.
The Effect of Sample Storage on Impedance and Optical Platelet Counts
Authors: Munro Roger C , Tranter Jeremy , Hart Laura , Jones Ruth , Benton Ann , Worthing Anita , Fitzpatrick Caroline , Al-Ismail Saad , Wakeman Lisa J 
Published: April 2020

Abstract
Introduction: Processing blood samples for full blood count may be delayed for a multitude of reasons. There is little published information on the effect of sample storage on platelet count. This study compares the effect of storing normal samples at room temperature (RT) and at 4°C prior to processing using different analysers with different technologies. Sysmex SF 3000 and XE2100 analysers are capable of providing impedance (IMP) platelet counts and the latter can also perform fluorescence based optical (OPT) platelet counts. The Coulter LH750 and the ABX Pentra DX120 use IMP technology with enhanced data extraction techniques and algorithms to eliminate interference.
Method: Intravenous blood samples for routine blood counts were taken into 4ml K2 EDTA Greiner Vacuette containers from 20 out-patients and the platelet counts measured at 0, 6 and 24 hrs on Coulter LH750 (IMP), ABX Pentra DX120 (IMP), Sysmex SF3000 (IMP) and Sysmex XE2100 (both IMP and OPT) blood count analysers. Samples were stored at RT. Samples were also processed on the XE2100 following storage at 4°C.
Results: Mean changes in platelet counts (n = 20) expressed as percentages of the original value at 0 hrs and paired t-test values of significance are shown in the Table. The most significant changes occurred with the XE2100 where the IMP value increased by 7% over 24 hrs but the OPT value fell by 5%. No significant changes occurred with the Coulter or the ABX analysers at 0 and 24 hrs or with the SF3000 at 6 hrs.
6 hr v 0 hr 24 hr v 0 hr % change p % change p Coulter RT + 0.19 0.873 −0.12 0.908 ABX Pentra DX120 RT + 0.02 0.985 + 0.57 0.380 SF 3000 RT + 1.25 0.281 + 2.52 &lt;0.05 XE 2100 Imp RT + 5.26 &lt;.05 + 7.06 &lt;0.05 XE 2100 Imp 4°C + 3.31 &lt;0.05 + 5.06 &lt;0.05 XE 2100 Opt RT −2.20 &lt;0.0 −5.32 &lt;0.05 XE 2100 Opt 4°C + 2.30 &lt;0.05 −0.40 0.750
EDTA exposure and the formation of membrane lesions during sample storage leads to changes in platelet structure, volume, function and shape due to decreased optical density and the beginning of degranulation. All these have a potential impact on the accurate assessment of platelet numbers. If these numbers are to be used accurately as thresholds for platelet transfusion or for monitoring progressive loss or increases of platelets associated with underlying pathology, it is vital that the pre-analytical limitations of current analysers are fully understood. The way in which discriminators are used on various analysers to make distinction between the characteristics of platelets and other blood cells in individual, stored samples will vary in their effectiveness.
Conclusions: Our data indicate that various combinations of pre-analytical sample storage and analyser technology should be considered as potential influences on the outcome of platelet counting in the clinical laboratory.
Studies on Platelet Storage Biomarkers: Effect of Different Protein Extraction Buffers on Platelet Gelsolin and B-Actin Profiling
Authors: Kulkarni Sandhya , Kannan Meganathan , Atreya Chintamani D 
Published: April 2020

Abstract
Platelet concentrates (PCs) are one of the blood cell components that play a major role in transfusion medicine. The shelf-life of PC is 5–7 days at 22°C. During storage PCs undergo certain morphological changes as well as physiological changes such as loosing of its surface glycoproteins, and reduced aggregation response to its agonists, which are collectively referred to as “storage lesions”. To date there is no in vitro “gold standard” that unequivocally serves as a surrogate marker to predict PC quality in vivo following transfusion. A comprehensive survey of reports in this field clearly indicate that there is some confusion with regards to the determination of which platelet protein or a panel of proteins could serve as potential markers associated with PC storage. Following a careful review of all available information, we recognized that perhaps one core issue to the observed discrepancies could be the buffer system that is being utilized by individual laboratories in extracting proteins in profiling protein alterations in PCs during storage. To address this, in the present study we analyzed two platelet proteins, b-actin and gelsolin-related 40 kDa protein. Profiles of these two proteins were previously reported by others to be altered during PC storage and hence implicated to be useful as biomarkers of platelet storage. By employing 3 different protein extraction buffer systems to isolate proteins from platelets at three storage intervals (Table 1), our results indicate that
b-actin and gelsolin 40 kDa protein levels do not appear to change with time in platelet storage and therefore may not be useful as platelet storage markers and Buffer systems do make a difference in extracting different platelet proteins for analysis, for example 40 kDa gelsolin was not extractable in buffer 3 (Table 1), suggesting that one should be careful in selecting a buffer system for platelet proteomics to have meaningful results.
Same level of 40 kDa Gelsolin Same level of b-actin Buffer system Day 0 Day 5 Day 16 Day 0 Day 5 Day 16 Buffer 1 Not identified Yes Yes Yes Yes Yes Buffer 2 Not identified Yes Yes Yes Yes Yes Buffer 3 Not identified Not identified Not identified Yes Yes Yes
The findings and conclusions in this abstract have not been formally disseminated by the Food and Drug Administration and should not be construed to represent any Agency determination or policy.
Effects of TAK-442, a Direct FXa Inhibitor, and Fondaparinux on Platelet-Associated Prothrombinase in the Balloon Injured Artery of Rats
Authors: Kawamura Masaki , Konishi Noriko , Katsuhiko Hiroe , Imaeda Yasuhiro , Fujimoto Takuya , Kubo Keiji 
Published: April 2020

Abstract
PCI patients routinely experience arterial injury during balloon angioplasty and stent deployment. At the injured arterial site, endothelial damage triggers platelet adhesion to the vascular wall and initiates platelet-associated prothrombinase complex (factor Xa [FXa]-factor Va-phospholipids-calcium) formation, resulting in the progression of thrombus formation. Recently, it was demonstrated that treatment with fondaparinux reduced the risk of death or recurrent myocardial infarction without an increase in the incidence of major bleeding compared to standard therapy in patients with STEMI. However, fondaparinux showed no superiority to placebo or heparin in patients undergoing PCI. On the other hand, small molecule direct FXa inhibitors, which are still in early stages of clinical development for the prevention of ACS, have been reported to strongly block reconstituted prothrombinase. Here, we compared the effects of TAK-442, a novel direct FXa inhibitor on platelet-associated prothrombinase activity, with fondaparinux. TAK- 442 and fondaparinux inhibited endogenous FXa activity in platelet-poor plasma from humans (IC50: 53 nM, TAK-442; 11 nM, fondaparinux) and rats (IC50: 32 nM, TAK-442; 19 nM, fondaparinux). When TAK-442 or fondaparinux were preincubated with free FXa, they each inhibited in vitro reconstituted prothrombinase activity, with IC50 values of 28 nM and 3.9 nM, respectively. In the absence of free FXa preincubation, TAK-442 retained this inhibitory effect (IC50, 51 nM); the inhibitory effect of fondaparinux, however, decreased dramatically (IC50, 1700 nM). In a rat model of balloon injury, thrombin activity on the surface of injured vessels increased to 3.3-, 20- and 5.7-fold that of intact aorta at 5 min, 1 hr, and 24 hrs after the injury, respectively. At 1 hr after the injury, 300 nM TAK-442 significantly (p&lt;0.025) inhibited platelet-associated thrombin generation on the surface of injured aorta by 99% (IC50, 19 nM), whereas the same concentration of fondaparinux showed no significant (p=0.076) inhibition (IC50, &gt;300 nM). These results highlight a limitation of fondaparinux in inhibiting platelet-associated prothombinase activity that leads to thrombus formation compared with TAK-442, and suggest that TAK-442 may be more effective in preventing arterial thrombosis in patients undergoing percutaneous coronary interventions.