bluuebunny/crossref_metadata_2025_split huggingface dataset:
Laser Tweezers Raman Spectroscopy Detects Differences between Normal Human Lymphocytes, Activated Lymphocytes and Leukemia.
Authors: Taylor Douglas Scott , Chan James W. , Zwerdling Theodore , Lane Stephen M. , Tuscano Joeseph , Huser Thomas 
Published: October 2019

Abstract
Raman spectroscopy is a laser-based analytical technique that enables chemical characterization of molecules in a sample. It is based on the inelastic scattering of photons by molecular bond vibrations. A Raman spectrum obtained from single cells is an intrinsic molecular fingerprint of the sample, revealing detailed information about DNA, RNA, protein, lipid, and carbohydrate content as well as macromolecular conformations. This technique is rapid, non-invasive and non-destructive. We have demonstrated that single-cell laser tweezers Raman spectroscopy (LTRS) reliably discriminates between normal human T- and B-lymphocytes and their transformed, Jurkat-T and Raji-B, counterparts. Herein the same technique is used to discern normal human T- and B-cells from T- and preB-leukemia cells. Also, demonstrated are Raman spectra changes that result from the activation of normal T-cells with a combination of TPA and ionomycin, and normal B-cells by cross linkage of surface immunoglobulin. T- and B-cells were isolated from healthy volunteers. T- and preB-leukemia cells were isolated from patient samples. Spectra are both compared directly and analyzed by principal component analysis. Significant Raman spectra differences in characteristic DNA and protein signals discriminate between normal cells and their leukemia counterparts. Human leukemia cells were indistinguishable from Jurkat-T and Raji-B cells. Changes in spectra that result from cell activation and proliferation only partially account for the spectra changes of leukemia cells. These data demonstrate the utility of LTRS to discern normal lymphocytes from both transformed cells and leukemia cells.
Laser Tweezers Raman Spectroscopy Detects Individual Neoplastic and Normal Hemotopoietic Cells.
Authors: Taylor Douglas Scott , Chan James W. , Zerdling Theodore , Lane Stephen M. , Ihara Ko , Huser Thomas 
Published: October 2019

Abstract
Current methods for identifying neoplastic cells and discerning them from normal counterparts are often non-specific, slow, biologically perturbing, or a combination, thereof. Here, we show that single-cell, laser-tweezers Raman spectroscopy (LTRS) averts these shortcomings and also permits mechanical manipulation of the single cell under investigation. LTRS is used to characterize the biomolecular Raman signature of both normal human lymphocytes and transformed Jurkat and Raji lymphocyte cell lines from single, unfixed cells in suspension. We demonstrate that single-cell Raman spectra provide a highly reproducible biomolecular fingerprint. Characteristic peaks, mostly due to different DNA and protein concentrations, allow for discerning normal lymphocytes from transformed lymphocytes with high confidence (p << 0.05). Spectra are also compared and analyzed by principal component analysis (PCA) to demonstrate that normal and transformed cells form distinct clusters that can be defined using just two principal components. The method is shown to have a sensitivity of 98.3% for cancer detection, with 97.2% of the cells being correctly classified as being normal or transformed. These results demonstrate the potential application of LTRS as a clinical tool, research instrument, and for cell sorting based on its intrinsic biomolecular signature, therefore eliminating the need for exogenous fluorescent labeling.
The Methylation Status of p21 Is Not Relevant in Adult Acute Lymphoblastic Leukemia Patients.
Authors: Gregorj Chiara , De Cave Fabiana , Ricciardi Maria R. , Scerpa Maria C. , Precupanu Cristina M. , Gervasoni Jacopo , Iannitto Maria L. , Fazi Paola , Gubbiotti Stefania , Specchia Giorgina , Cupri Alessandra , Ferrara Felicetto , Mondello Francesca , Rinaldi Ciro , Fioritoni Giuseppe , Cascavilla Nicola , Miraglia Eustachio , Annino Luciana , Audisio Ernesta , Mazza Patrizio , Peta Antonio , D’Arco Alfonso M. , Bigatti Catia , Tedeschi Alessandra , Foà Robin , Tafuri Agostino , Petrucci Maria Teresa 
Published: October 2019

Abstract
Methylation of CpG islands in the 5′ gene region is associated with transcriptional silencing of gene expression. The hypermethylation of tumor suppressor genes has been described in various tumor tissues, as in gastric and pancreatic cancer, as well as in acute myeloid leukemia, suggesting its potential role in tumorigenesis. Among the three members of the Kip/Cip family of cyclin dependent kinase inhibitors (CKI) p21, p27 and p57, little is known of their methylation status in hematological malignancies and contrasting studies have been reported on the role of p21 hypermethylation in the pathogenesis of acute lymphoblastic leukemia (ALL). The aim of our study was to analyze in primary blasts from adult ALL enrolled in the GIMEMA protocols 0496 and LAL2000 the methylation status of p21, defining in addition its protein expression by Western blot using the monoclonal antibody p21-WAF1 (Santa Cruz, CA). Primary samples from 81 untreated ALL patients were processed using a widely accepted method based on bisulfite modification of DNA, followed by the use of methylation-specific PCR assay (MSP). The human lymphoblastic cell lines (Jurkat, RPMI8866 and CEM), the myeloid cell line OCI-AML3 and normal peripheral blood lymphocytes (PBL) from 10 healthy donors were characterized by a consistent p21 promoter unmethylation (negative controls). In contrast, it was weakly methylated in the Raji cell line and strongly methylated in the Rael (Burkitt’s lymphoma) cell line (positive controls). This assay was further validate in vitro by SsI methylase. In the present study we analyzed 54 B-lineage ALLs, 25 T-ALLs and 2 biphenothypic leukemias; the mean WBC value at diagnosis was 125.6x109/L and 20 samples were Philadelphia chromosome positive. 71/81 of patients studied for p21 methylation were evaluated for response: 53 (74.6%) achieved complete remission (CR) after induction therapy, 8 (11.3%) patients were resistance and 10 (14.1%) died during induction therapy. DNA methylation was not observed in any of the adult ALL patients. p21 protein expression was found in OCI-AML3, Raji and RPMI8866 cell lines, while resulted negative in the Jurkat cell line and in normal PBL. Preliminary results obtained in the ALL samples showed that this protein was expressed in 8/29 (27.6%) cases. In summary, we demonstrated in a large number of primary ALL cases studied at presentation that the p21 gene is not methylated in this population and therefore that the status of p21 methylation does not play a role in the pathogenesis of adult ALL.
Marrow Plasma Levels of Stromal Cell-Derived Factor-1α in Patients with Hematopoietic Malignancies Might Be Associated with the Migration of Leukemic Cells.
Authors: Qian Jun , Lin Jiang , Chen Zi-xing , Chen Yong-jun , Cen Jiang-nong , Zhang Yong-ning , Ji Yong-hui , Qian Zhen 
Published: October 2019

Abstract
The chemokine, stromal cell-derived factor-1 (SDF-1), which is mostly produced by marrow stromal cells, has various effects on hematopoietic cell functions. Its crucial role in the survival, trafficking, and homing of hematopoietic stem cells and progenitor cells is currently established. Here we described the potential significance of bone marrow plasma levels of SDF-1α in patients with hematopoietic malignancies. The bone marrow plasma was obtained from 74 aspirate samples from 38 patients with acute myeloid leukemia (AML), 12 acute lymphoblastic leukemia (ALL) and 24 myelodysplastic syndrome (MDS). The concentration of SDF-1α was determined by a sandwich enzyme-linked immunosorbent assay (ELISA). The results revealed a significant negative correlation of marrow plasma levels of SDF-1 with peripheral white blood cells counts (r=−0.304, P<0.05). The patients with low or normal WBC count (<=10×109/L) had significantly higher SDF-1 levels than those with high WBC count (>10×109/L) (358.88±251.64 pg/mL vs 149.33±129.16 pg/mL, P<0.001). The correlation of SDF-1α levels with platelet counts was also observed (r=0.279, P=0.002). No statistically significant differences were observed in the marrow plasma levels of SDF-1α between normal and abnormal karyotypic groups. There was no significant difference between patients with AML and ALL (279.24±318.50 pg/mL vs 209.78±120.96 pg/mL, P>0.05), however, MDS patients had significantly higher SDF-1α levels (528.65±351.57 pg/mL) than AML and ALL patients (P<0.001). In the subtypes of AMLs, granulocytic leukemia has higher marrow plasma levels of SDF-1α (337.81±350.92 pg/mL, n=28) than mononuclear leukemia (104.94±96.12 pg/mL, n=7). Our results suggest that the migration of malignant leukemic hematopoietic cells, especially of mononuclear leukemic cells, might be associated with the marrow SDF-1α level.
Serum Dependency of t(8;21) AML Cell Line Is Associated with VEGF/VEGFR Pathway and Early Phosphorylation of Akt.
Authors: Imai Norikazu , Shikami Masato , Miwa Hiroshi , Hattori Akiko , Hiramatsu Akihito , Watarai Masaya , Satoh Atsushi , Itoh Masato , Imamura Akira , Mihara Hidetsugu , Nitta Masakazu 
Published: October 2019

Abstract
Most human leukemia cell lines are dependent on serum supplementation (usually fetal calf serum (FCS)), although the extent of serum dependency differs among each cell line. Kasumi-1, a t(8;21) AML cell line is one of the most serum-dependent cell lines. Since growth and survival of many leukemia cell lines are associated with phosphorylation of Akt, we examined the Akt phosphorylation by FCS treatment. In Kasumi-1, Akt was phosphorylated by culture with FCS in a dose-dependent manner, although no such Akt phosphorylation was observed in NB-4, a t(15;17) cell line. By FCS stimulation, Akt (Thr308, Ser473) was phosphorylated from 0.5 hr and the phosphorylation sustained until 48 hours in Kasumi-1. Then, we tested the effect of VEGF/VEGFR signaling in phosphorylation of Akt by FCS. The addition of VEGFR1/Fc and VEGFR2/Fc (which bind external VEGF and abrogate its function) inhibited the Akt phosphorylation from 2 hours until 10 hours, although the growth of Kasumi-1 was not inhibited. The addition of VEGFR2 kinase inhibitor (which inhibits internal VEGF signal) inhibited the Akt phosphorylation from 0.5 hr until 2 hours, and the growth of Kasumi-1 was greatly inhibited.
Taken together, it is suggested that serum dependency of Kasumi-1 is at least in part attributed to VEGF/VEGFR pathway. Then, both external and internal VEGF/VEGFR pathways work in Kasumi-1, which in turn phosphorylate Akt. However, blockade of only internal VEGF signal (by VEGFR2 kinase inhibitor) inhibit the early Akt phosphorylation (0.5 hr), which resulted in growth inhibition, indicating the importance of early Akt phosphorylation.
Results of Acute Myeloid Leukemia Treated with Triple-Drug Regimen Based on HA as Induction Chemotherapy and Its Relationship with Karyotype.
Authors: Wang Jianxiang , Mi Yingchang , Xue Yangping , Yu Wenjuan , Bian Shougeng 
Published: October 2019

Abstract
243 untreated de novo acute myeloid leukemia (AML) patients were treated with (homoharringtonine+AraC, HA) based induction therapy which composing of three chemotherapeutic drugs (HAD, HAM, HAA, HAE) in our hospital for recent 12 years. Complete remission (CR) rate, disease free survival (DFS) and overall survival (OS) of the patients were calculated. 184 patients who had karyotype results were divided into four groups according to SWOG (Southwestern Oncology Group) criteria. Differences of CR rate, DFS and OS of different groups were evaluated. The CR rate of all 243 cases was 77.4%, and 94.6% out of them were for 1~2 courses. Median DFS of the 188 CR patients was 28.5 (range from 1.0 to 153) months. DFS rates at 3 years and 5 years were 45.4%, 40.2% respectively. The median OS of 243 patients was 18.4 (range from 0.5 to 154) months. OS rates at 3 years and 5 years were 36.9%, 31.4% respectively. CR rate, DFS and OS of the different cytogenetic risk groups were also be analyzed. According to SWOG criteria, patients were classified into favorable, intermediate, adverse and unknown groups. CR rate, median DFS and OS were 97.8%, 87.4 months and 89.0 months in favorable group; 81.9%, 17.6 months and 22.3 months in intermediate group; 61.5%, 9 months and 11.5 months in the adverse group; 79.3%, 29.0 months, 19.9 months in the unknown group, respectively. The differences among the four groups were statistically significant. Multivariates analyses confirmed contribution of cytogenetics and courses of post-remission chemotherapy to DFS and OS. If we incorporated unknown group into the intermediate group, the CR rate is 80.6%. DFS rates at 3 years and 5 years were 40.5%, 38.1% respectively. OS rates at 3 years and 5 years were 40.3%, 37.4% respectively. We conclude triple-drugs induction regimens based on HA are highly effective in adult AML in China. Cytogenetics is the important prognostic factor. SWOG karyotype subtyping criteria is appropriate to our patients and unknown group can be incorporated into the intermediate group.
Possible Eradication of Minimal Residual Disease in Acute Leukemia.
Authors: Hijikata Yasuki , Maeda Yasuhiro , Yamaguchi Terufumi , Tanaka Miyako , Hirase Chikara , Shunsuke Takai , Morita Yasuyoshi , Matsuda Mitsuhiro , Kanamaru Akihisa 
Published: October 2019

Abstract
The complete remission (CR) rate in acute leukemia (AL) has been improved due to progressive chemotherapy. However, relapse frequently occurs as minimal residual disease (MRD). To eradicate MRD, immunotherapy using autologous cytotoxic T-lymphocytes has been available. Leukemia cells (LC) of the patients were stored frozen before chemotherapy. Furthermore, after achieving CR, an EBV-transformed lymphoblastoid B-cell line (LCL) was established in each case. To induce Cytotoxic T-lymphocytes (CTL), co-culture system was carried out in two types. One is co-culture of peripheral blood mononuclear cells (PBMCs) and original leukemia cells (LC), the other is co-culture of PBMCs and the patient’s own LCL. After co-culture for 5 days, cytotoxic activity was examined by FACS with PKH-26 staining. Interestingly, cytotoxicity against not only autologous LC but the patient’s own LCL was observed in almost cases. This phenomenon is called cross-killing. Moreover, regulatory T-cells (Treg) were detected in the effecter phase determined with FACS (CD4+ CD25+) and FOXP3 mRNA by RT-PCR. Enhanced cytotoxicity was observed in inverse relation to a low proportion of Treg. These results suggested that more cytotoxicity may be induced by deletion of the Treg population. In conclusion, cell therapy as an immunotherapy should be useful tool to improve the prognosis, and if stocked LC is not available, CTL activity must be induced by co-culture with the patient’s own LCL.
Acute Myeloid Leukemia (AML) in the Elderly. To Treat or Not To Treat: A Decision Analysis.
Authors: Kusminsky Gustavo D. , Rivas Maria M. , O’Flaherty Martin , Maggini Norberto J. , Pose Cabarcos Julio M. , Pinto Miguel Tezanos 
Published: October 2019

Abstract
AML is a disease frequently observed in elderly patients (pts) Treating this population with intensive chemotherapy or palliative treatment is a controversial decision. We analysed the available evidence with a decision model tree considering prognosis and comorbidities.
Method: A decision tree was designed to model significant events and prognostic variables that have an impact in survival at 5 years. The probabilities of this items were obtained from a literature review and for those items without literature support, assumptions were made by an expert opinion based on investigator’s consensus. Utility of each strategy was defined in an ordinal scale and the preference for each was calculated as the expected utility (utility x probability of the result)
Results: Probabilities were: unfavourable prognosis 95%, incidence of comorbidities 48%, death in induction without comorbidities 25%, response to induction in unfavourable prognosis 42%, response to induction in favourable prognosis 63%, adequate performance status after induction without comorbidities 43%, probability of finishing consolidation 64%, survival at 5 years in pts with unfavourable AML 11%, survival at 5 ys in pts with favourable AML 27%. The effect of comorbidities on survival to induction and adequate PS post induction was estimated as 50% higher than in pts without comorbidities. The survival at 5 ys in pts with no response or those who did not complete induction was considered equal to palliative treatment, and S at 5 ys in pts with comorbidities who finished consolidation 30% less than in those with no comorbidities. Utility for “not to treat” was 11, and for “to treat” was 6. The preference for “not to treat” was maintained for any modification of the survival to the induction chemotherapy independently of comorbidities. In sensitivity analysis, the only modification of the observed preference was that “To treat” strategy would be preferable only if the good PS after induction chemotherapy would be observed in 65% of pts or more. In conclusion, the preferred strategy as suggested by the model is “not to treat”, however many relevant questions are still unanswered, reinforcing the need to include these pts in clinical trials for better evidence to base the decisions.
Prospective, Randomized, Multicenter Phase III, Polish Adult Leukemia Group (PALG) Study Comparing DAF (DNR+AraC+Fludarabine), DAC (DNR+AraC+Cladribine), and Standard DA Regimen in Induction of Untreated Acute Myeloid Leukemia Patients - First Report.
Authors: Holowiecki Jerzy , Grosicki Sebastian , Kyrcz-Krzemien Slawomira , Kloczko Janusz , Sulek Kazimierz , Hellmann Andrzej , Skotnicki Aleksander , Kuliczkowski Kazimierz , Zawilska Krystyna , Robak Tadeusz , Dmoszynska Anna , Warzocha Krzysztof 
Published: October 2019

Abstract
The goal of this study is to compare the efficacy and safety of an original DAF regimen: daunorubicine (DNR) 60 mg/m2/d iv, days 1–3; cytarabine (AraC) 200 mg/m2/d ci, d 1–7, and fludarabine 25 mg/m2 2h inf. iv d 1–5 versus previously studied DAC regimen* (DNR, AraC, Cladribine), and versus standard DA in de novo acute myeloid leukemia (AML) patients below 60. Primary objective is complete remission (CR) rate after single course of induction and overall survival, secondary objectives - overall CR rate, toxicity, leukemia-free survival rate, assessment of lymphocyte subpopulations levels and survival in patients submitted to bone marrow allotransplantation (alloBMT) immediately after CR assessment.
Patients achieving CR and did not submit to alloBMT received two courses of subsequent intensive consolidation: 1) HAM (HD AraC, mitoxantrone) 2) HD AraC. In case of partial remission (PR) after the first induction course the same regimen was repeated, Patients with no remission (NR) or PR/NR after 2 induction courses were withdrawn with the study. We are planning to enrol to the study 600 patients in 3 years. Between 09.2004 and 07.2005, 147 adult AML patients, aged 48 (19–60)y, sex: male 57, female 90, treated in 16 co-operating Polish Adult leukemia Group (PALG) centres were randomised to either DAF (n=44), DAC (n=49) or DA (n=54) arm. PML/RAR alfa positive - FAB M3 cases were excluded. Both study groups were well balanced in respect of age, sex, FAB subtype, and WBC.
The final CR rate and CR rate after the first induction course equalled: for DAF 65% and 60%, for DAC 70% and 60%, and for DA 55% and 47%, respectively (p=NS).
The median times to ANC recovery > 0.5 G/L, and PLT >50 G/L in each arms were similar (22–26 d.) (p=NS). All patients developed WHO grade IV thrombocytopenia and agranulocytosis. The frequency and severity of infections, mucositis, vomiting, diarrhea, alopecia, polyneuropathy as well as of cardiac, liver or kidney failure were comparable in each treatment arms. Early death was noted in 6% (n=2) in DAF, in 9% (n=3) in DAC, and in 13% (n=5) in DA group, because of bacterial sepsis in every cases.
In conclusion, this original study proves that the addition of fludarabine to the standard DNR+AraC regimen (DAF) comparing to DAC and DA regimen is a potent antileukemic treatment without increased toxicity.
Clinical Investigation of G-CSF-Priming Regimen in Treatment of Relapsing, Refractory and Senile Acute Myeloid Leukemia.
Authors: Cai Zhen , Han Xiaoyan , Huang He , Xie Wanzhuo , Zheng Weiyan , Lin Maofang 
Published: October 2019

Abstract
Objective:To evaluate the clinical outcome of granulocyte colony-stimulating factor (G-CSF) priming regimen in treatment of refractory, relapsing and senile aute myeloid leukemia.
Materials and Methods:16 patients, 10 men and 6 women ranged in age from 15 to 78 years (median 48), with refractory, relapsing or senile acute myeloid leukemia were treated in our hospital between March 2002 and January 2004. All patients were diagnosed with MIC (morphologic, immunologic, cytogenetic) classification. The control group consists of 18 patients (12 men and 6 women, median age 48) with refractory, relapsing or senile acute myeloid leukemia treated in our hospital between 1999 and 2002. All of them were also diagnosed with MIC. All patients enrolled were treated with G-CSF-priming regimen consisting of low-dose cytosine arabinoside (Ara-C 10mg.m−2.12h−1, subcutaneously,day1–14), aclarubicin (Acla5-7mg.m−2.d−1, continuous intravenous infusion, day 1–8; or 10–14mg.m−2.d−1, continuous intravenous infusion, day1–4) or homoharringtonine (HHT 1mg.m−2.d−1, continuous intravenous infusion, day1–14), and concurrent use of G-CSF (100ug.m−2.12h−1, subcutaneously, day 1–14). In the course of chemotherapy, blood routine examinations were made every other day, and G-CSF was suspended while WBC>20x109/L. Chemotherapy intermission was 14–21 days. If two courses were of no effect, then the investigation was terminated. Patients in the control group received intensive chemotherapy with medium /high dosed Ara-C (1-3g.m−2.12h−1× 3–5d), combined with mitoxantrone or daunorubicin (DNR) or etoposide (vp-16).Statistics:Enumeration data was analyzed by Chi-square test.
Results: All patients accomplished the chemotherapy, with no treatment related death. 9 of the 16 patients achieved complete remission. The CR rate was 56.25%, compared with 22.2% of the control group (Χ2=4.15, P<0.05). However, the total effective rate (68.75%) was not statistically higher than the control group (44.4%) (Χ2=2.03, P>0.05). Therapy related mortality (0.0%) was apparently decreased in priming group than in control group (38.9%) (Χ2=7.84, P<0.01).
Conclusions:Our study shows that the priming regimen is effective and safe to refractory, relapsing, secondary or senile acute myeloid leukemia. Further studies are necessary to evaluate the long-term effects of this new therapeutic regimen in AML.