bluuebunny/crossref_metadata_2025_split huggingface dataset:
Polymorphisms in Genes of Folate Metabolism and Response to High-Dose Methotrexate in Patients with Primary Central Nervous System Lymphoma.
Authors: Knop Stefan , Loeffler Juergen , Steffens Michael , Guenther Markus , Korfel Agnieszka , Weller Michael , Hebart Holger , Einsele Hermann , Herrlinger Ulrich 
Published: October 2019

Abstract
The antifolate methotrexate (MTX) was shown to be the single most-effective agent in first-line treatment of patients with primary central nervous system lymphoma (PCNSL) when given intravenously in high doses. MTX inhibits 5,10-methylentetrahydrofolate reductase (MTHFR) as well as thymidylate synthase (TS) in target cells what eventually results in decreased DNA synthesis. Intracellular uptake of both folate and MTX is mediated by the reduced folate carrier (RFC). Genetic polymorphisms for all three proteins were described: a C to T base transition at nucleotide 667 (C677T) and an A to C transition at 1298 (A1298C) of MTHFR; a 28-base pair (bp) tandemly repeated sequence polymorphic in the numbers of the repeat (2R/2R; 3R/3R; and 2R/3R) in the 5′-UTR of TS; a 6bp insertion-(+6bp)/deletion(−6bp)-polymorphism in the 3′-UTR of TS; and a polymorphism at nucleotide 80 (G80A) of RFC. We hypothesized that these polymorphisms may be directly linked to response, survival and toxicity in patients with PCNSL who are treated with single high-dose MTX. We prospectively collected blood from 152 subjects who were enrolled into a German multicenter PCNSL trial. They were scheduled to receive HD-MTX 4 g/m2 every 2 weeks. Genomic DNA was extracted from mononuclear cells using QIAGEN DNA Blood Mini Kit®. Genotyping of the two MTHFR (C677T; A1298C) and the RFC (G80A) single nucleotide polymorphisms (SNPs) was performed by melting point analysis with Master Hybridization Probes® using the Light Cycler® technology. Genotyping of the TS 28bp tandem/triplet repeats in the 5′-UTR was performed by conventional PCR followed by high resolution agarose gel electrophoresis. The TS +6bp/−6bp polymorphism in the 3′-UTR was analyzed with PCR, followed by restriction enzyme digest with draI and agarose gel electrophoresis. The fragment sizes were 70 bp and 88 bp for the 6 bp insertion allele and 152 bp for the 6 bp deletion allele. Allele frequencies for heterozygosity and for homozygous mutations of the two MTHFR and the RFC SNPs as well as of the two TS polymorphisms were as expected for a Caucasian population. For the currently 120 evaluable patients logistic regression analysis, Armitage Trend test and Kaplan-Meier survival curves were performed to analyze the role of the polymorphisms as potential predictors for MTX response. Up to now, we only found a trend for an association between response to treatment and survival for the 2 MTHFR mutations. We will analyze haematological and non-haematological toxicities which will be presented at the meeting with updated results for survival and response in the patients who will have completed treatment until then.
Monitoring CML28 mRNA Levels in Patients before and Post HSCT by Real-Time Quantitative RT-PCR.
Authors: Zhang Donghua , Dai Min , Zhou Hongsheng , Wang Yaya , Zhang Lu , Zhang Li , Wang Bin , Cao Wenjing 
Published: October 2019

Abstract
A SYBR Green I real-time quantitative RT-PCR method was established for investigating the correlation between CML28 mRNA expressing levels and relapse of leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). pcDNA3.1HisA-CML28 plasmid had been constructed as the standard template. Serial monitorting of CML28 mRNA levels by SYBR Green I real-time quantitative RT-PCR technique was performed in 14 patients, including 10 patients with CML and 3 patients with AML, 1 patient with Ph+ ALL. The sensitivity of the established method was at 10−4 level, with interassay variation and intraassay variation of standard samples both < 10%. The CML28 was highly expressed in AML and CML-BP or AP. In newly diagnosed group, CML28 was (6.58±2.34)×10−2. In pre-conditioning regimen group was (2.19±0.32)×10−2, in group that 1 month after allo-HSCT was (1.35±1.28)×10−2, in group that 3 months after allo-HSCT was (4.57±6.39)×10−3. CML28 can be detected 3months after HSCT in 1 patient with CML-CP and 3 patients with CML-AP or BC. 2 of them with low level (<2×10−2) survived without relapse, the other 2 patients with high level (>2×10−2) relapsed within one year,1 died and1 received the second time allo-HSCT, CML28 level decreased rapidly after HSCT, but still higher than 2×10−2 and relapsed again. CML28 mRNA level was obviously correlated with the development of diseases. Serial quantification of CML28 mRNA levels were necessary for allo-HSCT recipients, and more informative than a single detection. Use of this assay to evaluate MRD in the patients performed allo-HSCT was helpful for predicition of relapse.
SDF-1-CXCR4 Signaling and Downregulation of CD26/Dipeptidyl-Peptidase IV Are Involved in Skin-Homing of Sezary Cells.
Authors: Russo Giandomenico , Scala Enrico , Bresin Antonella , Caprini Elisabetta , Picchio Maria Cristina , Remotti Daniele , Ragone Gianluca , Nasorri Francesca , Frontani Marina , Cristofoletti Cristina , Arcelli Diego , Lombardo Giuseppe A. , Baliva Giannandrea , Napolitano Monica , Narducci Maria Grazia 
Published: October 2019

Abstract
Sezary Syndrome (SS) is a rare form of Cutaneous T-Cell Lymphoma (CTCL) characterised by a distinct metastatic pattern mainly involving skin and blood. Chemokine and chemokine receptors have been implicated in the spreading process of many cancers including various forms of non-Hodgkin T-cell lymphomas (NHL). In this study we report that chemokine receptor CXCR4 is over-expressed by both circulating and skin-homing neoplastic T-lymphocytes of SS patients and is functionally active as demonstrated by the migration of freshly isolated Sezary (SzS) cells along the chemical gradient of its natural ligand SDF-1. To shed light on the regulation of CXCR4/SDF1 interaction, we also investigated the enzymatic activity of CD26/dipeptidylpeptidase IV (DPPIV) since SDF-1 is efficiently inactivated by CD26, in physiological condition.. This is of particular relevance because one of the hallmark of the circulating SzS cells is the loss of CD26 from the cell surface. We first demonstrated that the CD26 negative phenotype is similarly maintained also in the skin-homing neoplastic T lymphocytes; we then observed that the addition of exogenus soluble CD26 reduces the migratory response of SS cells to SDF-1 whereas the inhibition of the CD26 peptidase activity in Hut78, a CD26-positive CTCL cell line, enhances the SDF-1-induced migration of these cells. We finally showed that SS individuals exhibit a reduced activity of the soluble CD26 as revealed by the measurements performed on the patients derived plasma. Our findings suggest that the SDF-1-CXCR4 axis could play an important role in skin homing of SS through the regulatory activity of CD26.
Impact of RNA Stabilization on Minimal Residual Disease Assessment in Chronic Myeloid Leukemia.
Authors: Thörn Ingrid , Olsson-Strömberg Ulla , Ohlsen Cecilia , Jonsson Anna-Maria , Klangby Ulf , Simonsson Bengt , Barbany Gisela 
Published: October 2019

Abstract
Background and objective: Accurate quantification of BCR-ABL mRNA is of critical importance for managing chronic myeloid leukemia (CML) patients receiving Imatinib therapy. RNA degradation thus constitutes a potential problem for laboratories quantifying minimal residual disease (MRD). Patient samples with long transport times between the hospital and the analyzing laboratory may be subject to RNA degradation with a corresponding loss in sensitivity and possible generation of false negative results. Recently, RNA preservation systems have been developed in order to improve RNA stability.
Design and methods: The aim of the study was to investigate the performance of the PAXgene Blood RNA Kit in CML follow-up peripheral blood samples and compared the results to unstabilized parallel Trizol extracted samples. The different sample processing methods were evaluated by real-time PCR.
Results: RNA isolated with the PAXgene system gave a superior yield per ml blood than with the routine Trizol extraction method. However, although of comparable quality, the RNA did not PCR-amplify as efficiently compared to equal amounts of RNA from routinely processed samples. Therefore, RNA processed with the PAXgene system showed to decreased sensitivity for MRD detection, resulting in false negatives. The sensitivity was comparable to samples processed routinely 20–30 hours after phlebotomy.
Interpretation and conclusions: We conclude that routinely processed, i.e. unstabilized, peripheral blood that reaches the laboratory and is processed within 30 hours is preferable for MRD detection. Optimal results were achieved with fresh samples processed within 5 hours with the Trizol method. However, RNA stabilization may be useful if sample transit is expected to exceed 30 hours.
CD38 Expression in B-Cell Chronic Lymphocytic Leukemia: Association with Clinical Presentation and Laboratory Features in 64 Newly Diagnosed Patients.
Authors: Matias Kleber , Freire Ana Dulce , Matias Carolina , Neves Maria Amélia , Machado Cintia 
Published: October 2019

Abstract
Background: B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the progressive accumulation of a neoplastic clone consisting of CD5/CD19/CD23/surface immunoglobulin cells. Despite this homogeneity in phenotype, B-CLL can follow either an indolent or a progressive course. CD38 is a transmembrane glycoprotein expressed on the surface of leukemic cells in a significant percentage of patients with B-CLL. Recent studies suggest that CD38 expression in CLL is a reliable prognostic marker, leading to an unfavorable clinical course with a more advanced stage of disease, poor responsiveness to chemotherapy and a shorter survival rate, along with others markers like ZAP-70 that reflect more accurately the mutagenic status of VH gene region, unfortunately not yet available in clinical practice routinely.
Objectives: To assess the association of CD38+ expression with clinical and laboratory parameters at diagnosis in patients with B-CLL.
Patients and Methods: CD38 expression was analyzed in 64 unselected newly diagnosed B-CLL patients from January 2003 to May 2005 seen at the Hematology Center of Pernambuco-Brazil (HEMOPE). According to Damle et al. patients with 30% or more B cells expressing CD38 were considered positive(CD38+), and those with less than 30% were considered negative(CD38−). Various patient characteristics were studied including age, sex, Binet stage, hemoglobin, ß2 microglobulin and lactate dehydrogenase levels in the serum. Statistical differences between each group (CD38+ vs CD38−) were analyzed using χ2 tests for categorical variables and Student’s t-tests for continuous variables.
Results: Thirty-six patients (56.25%) were CD38+ and 28 (43.75%) were CD38-. Their median age at diagnosis was 64 years (range, 39–83) and the male to female ratio was 1.46:1. There were no differences between age, sex, hemoglobin and ß2 microglobulin levels in the CD38+ and CD38- groups. ß2 microglobulin level, however, was not available for assessment in all patients. Lactate dehydrogenase level was significantly higher in the CD38+ group (p=0.007). It was observed a trend toward a higher Binet stage (B or C) in the CD38+ compared with CD38- group (63.4 vs. 36.6% respectively), although not statistically significant (p=0.09) possibly because of the small number of patients studied.
Conclusion: CD38 expression was associated with a higher lactate dehydrogenase level and a tendency for more aggressive clinical stage. CD38 evaluation is a measurable biological parameter that is not as subjective as the evaluation of some clinical parameters and should be considered a stantard clinical test for newly diagnosed B-CLL patients.
Cytogenetic Classification Systems and Overall Survival Following Bone Marrow Transplant (BMT) for Acute Myelogenous Leukemia (AML).
Authors: Kalaycio Matt , Sekeres Mikkael , Sobecks Ronald , Rybicki Lisa , Pohlman Brad , Advani Anjali , Kuczkowski Elizabeth , Bolwell Brian 
Published: October 2019

Abstract
Risk-adapted therapy for AML in first complete remission generally calls for allogeneic BMT for patients with poor risk cytogenetics. However, poor risk cytogenetics is defined differently in commonly-applied classification schemes. We hypothesized that differences in cytogenetic classification might result in differences in survival after BMT. From September 1991 to December 2003, we treated 47 patients with AML in first complete remission with high-dose busulfan-containing preparative regimens and an HLA-matched sibling BMT. The median age was 42 years (range 18 to 60). At the time of diagnosis, 35 patients had either a normal karyotype or at least one clonal abnormality. Cytogenetic analysis was unavailable for the other 12 patients. The 47 patients were then classified according to the SWOG/ECOG (Blood96:4075, 2000), MRC (Blood92:2322, 1998), or CALGB (Blood100:4325, 2002) cytogenetic classification systems and analyzed for overall survival. With a median follow-up of 4.9 years, 3 year overall survival for various cytogenetic risk groups are displayed in the table:
SWOG/ECOG CALGB MRC Intermediate risk 69% 63% 74% Poor risk 43% 42% 18% Unknown risk 45% 52% 45%
Concordance indices were 0.58 for CALGB, 0.60 for SWOG, and 0.66 for MRC, demonstrating a slight superiority of the MRC system. Consistent with reports of large, prospective randomized trials of BMT for AML in first remission, patients with poor risk cytogenetics have worse overall survival compared to patients with intermediate risk cytogenetics. Overall survival appears to be different for patients classified as poor risk by MRC criteria. Larger, prospective studies are needed to confirm this observation, but our results suggest that standardized, international cytogenetic risk criteria are needed to develop risk-adapted strategies for the treatment of AML.
Simplifying Complex Microarray Data To Derive Gene Expression Profiles Which Identify Childhood Acute Lymphoblastic Leukaemia Patients at Risk of Relapse.
Authors: Guo Dachuan , Fong Alex , Lail Andy , O’Sullivan Maree , Stone Glenn , Kiiveri Harri , Henry Michael , Stephan Dietrich , Dalla-Pozza Luce , Catchpoole Daniel R. 
Published: October 2019

Abstract
The optimal treatment of patients with childhood acute lymphoblastic leukaemia (ALL) depends on establishing accurate diagnosis. Our investigations seek to strategically develop the application of microarray gene expression profiling to identify ALL patients with clinically homogenous presentations but which may respond differently to established treatment regimens. We have determined the gene expression profiles of ALL bone marrow (BM) samples taken from patients at diagnosis. Data analysis has focussed on the use of a novel and innovative statistical technology, Gene-RaVE. This series of patent protected algorithms builds a multinomial regression model using Bayesian variable selection. Gene-RaVE leads to the generation of a parsimonious and simple diagnostic gene expression signature, but which provides increased predictive ability over current analysis approaches. We describe our analysis of both Affymetrix (HU133A) and cDNA (10.5K) microarray gene expression profiles generated from diagnostic BM from >100 ALL patients covering the broad ALL subtypes including T and B lineage as well as T cell lymphoma leukaemia. Comparison of gene expression data failed to identify clearly distinguishing profiles between patients identified as ‘standard risk’ from ‘medium risk’ according to BFM95 clinical criteria. Gene expression profiles from a cohort of ALL patients, identified as ‘standard risk’ at diagnosis, were compared on the basis of their overall clinical outcome: relapse within 2 yrs vs non-relapse. Using a range of analyses including t-test, Gene-RaVE, discriminant analysis approaches and principle component analysis, we have discovered that small subsets of genes (<10), all of which included Nedd4BP3 and Ribosomal Protein L38 (RPL38), can be used to distinguish the two outcome groups. Subsequent validation using real time PCR supports the increase in Nedd4BP3 expression in standard risk patients which do not respond well to established treatment regimens. The Gene-RaVE algorithm also provides a generic framework for survival analysis. This approach indicates that the expression of these Nedd4BP3, RPL38 and inositol 1, 4, 5-triphosphate receptor, type 2 can be used to build a survival ‘index’ which correlates with the time to a relapse event in standard risk childhood ALL patients. Our results are suggestive of a way forward in the development of an informative, yet efficient diagnostic tool for this childhood malignancy using microarray gene expression analysis technology.
A New Predictive and Prognostic Marker for De Novo AML: Peripheral Blood CD34 (pCD34) Count at Recovery Following Remission Induction (RI) Therapy (Supp. by Ankara University-2003-0809114).
Authors: Arat Mutlu , Topcuoglu Pervin , Yuksel Meltem K. , Dalva Klara , Gultekin Gulhis , Gurman Gunhan , Arslan Onder , Ozcan Muhit , Beksac Meral , Konuk Nahide , Uysal Akin , Koc Haluk , Ilhan Osman 
Published: October 2019

Abstract
In EORTC-LCG AML-10 study patients with low CD34 mobilization profile showed a better outcome (Leukemia,2003,17:60-7). We hypothesized that patients with low pCD34 at the recovery period after RI therapy with de novo AML should have a better clinical outcome or vice versa and launched this prospective study. Between Jan 2002 and Oct 2004, 40 patients with AML (median age: 40 yrs, 17–60; 22M/18F) were enrolled to this study. Using flow cytometrical method CD34-expressing cells were measured in peripheral venous blood samples once WBC was between 1x10e9/L to 3x10e9/L at the recovery period after first RI therapy.
Results: The median time for estimation of pCD34 was 23 days (7–35) after RI. The pCD34 cell count was lower in patients responding to first RI therapy (12x10e6/L vs 68x10e6/L, p=0.011). We observed a weak, but statistically significant positive correlation between CD34 expression of the blasts at diagnosis and pCD34 at recovery period (p=0.048, r2:0.322). We could not show any significant negative impact of CD34 expression of the blasts at diagnosis and response to first RI (p=0.059). Only two of 8 patients not responding to the 1st RI achieved a complete remission after re-induction. The two-year probability of OS, LFS, relapse incidence and mortality rates were 47.1%, 37.2%, 55% and 42.5%, respectively (fig 1,2). If we set up a cut-off value of 18x10e6/L for pCD34, we did not observe any impact of pCD34 on relapse incidence but the mortality rate was significantly increased in patients with high CD34+ cell count (p=0.025).
Conclusion: We were able to show a positive impact of pCD34 estimations after first RI in AML patients on both remission and mortality rate. AML Patients with lower pCD34 after 1st RI tends to have a higher hematological remission rate and lower mortality. This impact of pCD34 as a predictive and prognostic marker in AML should be verified in large cohorts using multivariate analysis.
Table 1: Distribution of risk factors at diagnosis according RI response Variables 1stCRafter1stRI (n=32 ) NoCR after 1stRI (n=8 ) P *p<0.05; Abbr: NS: Non-Significant; ND: Not-Done pCD34 (x10e6/L) 12.4 (0–268) 68.6 (4.4–3820) 0.011* CD34 expr at dx 12% (0%–94%) 44% (9–89) 0.059 Age, years 38.5 (17–57) 43 (20–60) NS WBC at dx (x10e9/L) 11.85 (0.7–175.0) 5.10 (2.7–114.7) 0.934 EM involvement (Y/N) 7/25 0/8 NS Table 2: The outcome of AML patients according to median pCD34 at recovery Variables pCD34<Med 18x10e6/kg (n=20 ) pCD34 ≥ Med 18x10.6/kg (n=20) p CR1 (Y/N) after 1st RI 19/1 13/7 0.044* Relapse incidence 44% 54.5% NS Mortality rate 25% 60% 0.025* DFS 46.19%±12.6% 29.2%±10.4 NS OS 64.95%±13.2% 35.6%±11.3 NS
Figure Figure
Complementary Diagnostic and Prognostic Value of Parallel Karyotyping and Molecular Diagnostics in Patients with Acute Leukemias.
Authors: Ejduk Anna , Majewski Miroslaw , Solarska Iwona , Makuch-Lasica Hanna , Kania Iwona , Nowak Grazyna , Seferynska Ilona , Konopka Lech , Warzocha Krzysztof 
Published: October 2019

Abstract
Chromosomal aberrations analyzed by cytogenetic and molecular methods are major prognostic factors determining treatment options in patients with acute leukemias. The aim of this study was to compare cytogenetic and molecular methods as diagnostic and prognostic tools in patients with acute leukemias. Sixty one previously untreated patients with acute leukemias (AML - 43, ALL - 18) were studied for the presence of chromosomal translocations and corresponding fusion genes [AML1-ETO t(8;21), CBFB-MYH11 inv(16), PML-RARA t(15;17), BCR-ABL p190, p210 t(9;22), MLL-AF4 t(4;11), TEL-AML1 t(12;21), E2A-PBX1 t(1;19), SIL-TAL1 del(1)] using both molecular (RT-PCR, nested PCR) and cytogenetic (GTG) methods. Molecular diagnostics was performed from RNA isolated from bone marrow samples according to BIOMED-1 protocol. Cytogenetic studies were carried out with classical GTG and FISH methods. G-banded mitoses of bone marrow specimen were analysed according to ISCN. Chromosomal aberrations were found in 32.8% patients using GTG method while the parallel molecular tests revealed related fusion genes in 50.8% patients. In 8.5% patients cytogenetic analysis was not performed because of lack of metaphases in cultured cells. All cytogenetic aberrations found in GTG were also confirmed in RT-PCR. Stratification into cytogenetic risk groups was performed after applying combined analysis of karyotyping and molecular tests. Low cytogenetic risk group consisted of 32.8% patients including 11.5% diagnosed with GTG and additionally 21.3% patients after applying molecular tests. The intermediate and high cytogenetic risk group consisted of 32.8% and 34.4% respectively using combined cytogenetic and molecular diagnostics. In low cytogenetic risk group, 85% of patients achieved complete remission (CR), early deaths were found in 15% and none of the patients presented primary chemotherapy resistance. In intermediate risk group CR were obtained in 80%, chemoresistance in 10% and early deaths were observed in 10% of patients. In high cytogenetic risk group, CR were achieved in only 23.8% and chemoresistance occured in 76.2% of the patients. In conclusion we suggest that molecular and cytogenetic tests are complementary methods and should be used in parallel in the initial diagnosis of patients with acute leukemias. This seems to be critical for obtaining the accurate diagnosis, cytogenetic risk assessment and choosing an optimal treatment options.