bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mobilization, Harvesting and Selection of Peripheral Blood Stem Cells in Patients with Autoimmune Diseases Undergoing Non-Myeloablative Autologous Hematopoietic Stem Cell Transplantation.
Authors: Statkute Laisvyde , Verda Larissa , Oyama Yu , Villa Marcelo , Shook Thomas , Clifton Ronnie , Krosnjar Nela , Quigley Kathleen , Yaung Kimberly , Weppner Catherine , Burt Richard K. 
Published: October 2019

Abstract
We have analyzed peripheral blood stem cell (PBSC) mobilization, harvesting and selection properties in 128 patients with severe autoimmune diseases undergoing non-myeloablative autologous hematopoietic stem cell transplantation (HSCT) (50 patients with systemic lupus erythematosus (SLE), 43 - with multiple sclerosis (MS), 15 - with Crohn’s disease (CD), 8 - with scleroderma (Scl), and 12 - with others). Female/male ratio and mean age (range) were 90/38 patients, and 34 (14 to 59) years old, respectively. Mobilization regimen included cyclophosphamide 2g/m2 and G-CSF 10 mcg/kg (except for SLE patients 5 mcg/kg). Forty one patients underwent stem cell collection using Baxter CS300, 78 patients - Spectra, and for 9 patients both apheresis machines were utilized. The mean number of aphereses was 1.8 (range 1–10). Patients with SLE required the largest number of apheresis sessions (mean 2.4), comparing to patients with CD (mean 1.9), Scl (mean 1.4), MS (mean 1.3). Five patients additionally required bone marrow harvest for collection of adequate numbers of stem cells. One patient failed to reach CD34+ cell number of 1.0x106/kg, therefore did not proceed to HSCT. The mean number of CD34+ cells in each apheresis unit was 6.07+−6.96x106/kg (the highest of 9.22+−8.52x106/kg in patients with MS, and the lowest of 3.93+−4.48x106/kg in patients with SLE). Ninety eight patients underwent stem cell selection with CEPRATE SC (N=18), Isolex 300iv1.12 (N=2) or Isolex 300iv2.5 (N=78) stem cell concentrator. The mean purity of selected products was 74.3% (the highest of 81.1% attained in patients with Scl); mean recovery of CD34+cells was 61.2%. T cell reduction by average of 3.7 logs was achieved. The mean number of infused CD34+ cells was 7.24+−5.5x106/kg. The highest mean number of CD34+ cells/kg were infused to patients with MS (9.04+−6.74x106/kg), the lowest - to patients with SLE (5.78+−4.13x106/kg). We found a moderate positive correlation between peripheral blood (PB) CD34+ cells/ul and PB WBC/ul (R=0.34, p<0.05), PB platelets/ul (R=0.51, p<0.05) and a strong positive correlation between PB CD34+ cells/ul and the number of CD34+ cells/kg/apheresis (R=0.67, p<0.05). A weak positive correlation was observed between the number of infused CD34+cells/kg and faster WBC engraftment (ANC>500) and platelet engraftment (platelet count>20K). There was no toxicity observed in our patient population during peri-mobilization period except for 1 patient with SLE who died of disseminated mucormycosis 1 week after stem cell collection. Mobilization and selection of PBSC are safe and efficient in patients with severe autoimmune diseases undergoing non-myeloablative autologous HSCT.
Quantification of Imatinib in Human Plasma by High Performance Liquid Chromatography-Tandem Mass Spectrometry: Applications to Therapeutic Drug Monitoring of Patients with Chronic Myelogenous Leukemia.
Authors: Mahon Francois-Xavier , Picard Stéphane , Titier Karine , Nicolini Franck , Ducint Dominique , Moore Nicolas , Marit Gerald , Berthaud Patrice , Molimard Mathieu 
Published: October 2019

Abstract
Imatinib (Gleevec® or Glivec®) is a competitive inhibitor of protein tyrosine kinase is currently used for the treatment of chronic myeloid leukaemia (CML), Philadelphia chromosome-positive acute lymphoblatique leukemia and for other malignant pathologies.
Although there is no clearly defined therapeutic window, the imatinib plasma quantification seems interesting to evaluate patient compliance to daily oral therapy, drug-drug interactions or pharmacokinetic/pharmacodynamic relationship. The aim of this study was to develop a simple and accurate liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the imatinib quantification, adapted to routine applications. After a liquid-liquid extraction, the imatinib and its deuterated internal standard were eluted on a XTerra® RP18 column with a gradient of acetonitrile-ammonium formiate buffer 4 mM pH 3.2. Imatinib was detected by electrospray ionisation mass spectrometry with multiple reaction monitoring mode. The calibration curves were linear over the range 10–5000 ng/ml. The limit of quantitation was set at 10 ng/ml. The bias was lower than 8 %. Intra-day and inter-day precisions were lower than 8 %. The extraction recovery was higher than 90 %.
The monitored population was defined as follows: patients treated for CML and receiving imatinib with one daily dose of 300, 400 or 600 mg. Blood samples were collected at steady state, 23 (±3) hours after drug administration, and were centrifuged at 4500 rpm for 5min. As shown on the figure 1, all patients treated with 600mg have a concentration more than 500 ng/ml which corresponds to the efficient concentration of imatinib according to the previously in vitro studies (i.e. the target plasma concentration required to induce the death of leukemic cells). Five patients - previously known as not having an efficient hematologic response or cytogenetic response showed an imatinib level lower than 500 ng/ml Two of these patients were not compliant and three patients were coadministered with enzymatic inducers: phenytoïn or phenobarbital or oxcarbazepine This method is simple, adapted to routine application and allows accurate therapeutic monitoring of imatinib. It can be used to evaluate patient adherence to daily oral therapy, drug-drug interactions or pharmacokinetic/pharmacodynamic relationship. In conclusion, regarding the definition imatinib resistance in CML, it seems necessary to take in account the value of the imatinib concentration. A study of correlation with the cytogenetic and molecular response is in progress.
Figure Figure
Use of Direct Sequence PCR for ABl Kinase Mutations in Patients with CML Blast Crisis, Treated Prior to the Availability of Imatinib Therapy.
Authors: Ghorashian Sara , Babb Anna , Khorasan Jamshid , Kaeda Jaspal , Marin David , Apperley Jane 
Published: October 2019

Abstract
We identified all patients who developed blast crisis before the advent of imatinib therapy and in whom stored marrow samples were available from a database of CML patients in a major teaching hospital. mRNA was extracted and cDNA synthesized successfully in 20 of 22 cases. The cDNA was subjected to real time PCR for quantification of BCR-ABL transcripts. The ABL allele not involved in the t(9;22) translocation was excluded from mutational analysis by subjecting the cDNA to nested PCR. This was achieved by amplifying from exons 13 and 10 of the BCR and ABL moieties of the BCR-ABL fusion gene using primers B2A and JAMR, respectively. The resulting amplicon was then subjected to nested PCR using primers, NTPB+ and NTPE-, sited within exons 4 and 10 of the ABL gene. The nested PCR yielded an 863 bp fragment in length containing the entire BCR-ABL kinase domain. To test for successful amplification an aliquot of the PCR products were electrophoresed through 2.0% agarose gel. In all cases, a single amplicon was observed and the PCR reaction subjected to magnetic purification. The purified amplicon were then sequenced by Sanger’s dideoxy chain termination reaction using Big-Dye ABI 310 sequencer (Applied Biosystems, Foster City, USA). In each case, the sequence obtained was compared with the published ABL type 1a sequence, Genbank M14752, using Blast 2 software. No mutations were found in any patient. This suggests the absence of a mutated kinase in a proportion of greater than approximately 30% of the total BCR-ABL kinase, due to the limit of detection of the technique. Our results suggest that the predominant form of BCR-ABL in the setting of blast crisis (in the absence of imatinib) is wild type. This goes against theories that kinase mutations confer an intrinsic ‘gain of function’ or a proliferative advantage to a CML clone. Instead, it is likely that mutant forms of BCR-ABL which are relatively resistant to imatinib binding gain prominence through positive selection with the presence of imatinib within the cell milieu.
Enforced STAT3 and PU.1 Expression Activates Type I Interferon-Producing Cell and Dendritic Cell Differentiation Programs in Megakaryocyte/Erythrocyte Progenitors.
Authors: Onai Nobuyuki , Onai Aya , Tussiwand Roxane , Lanzavecchia Antonio , Manz Markus G 
Published: October 2019

Abstract
A standing question in early hematopoiesis is whether cytokine signaling is sufficient to induce cell fate decisions. Previously, we have reported that enforced expression of human Flt3 in Flt3 negative megakaryocyte/erythrocyte progenitors (MEPs) rescues their interferon producing cell (IPC) and dendritic cell (DC) developmental potential: Human Flt3-signaling in MEPs leads to up-regulation of IPC, DC, and myelomonocytic development affiliated genes such as, STAT3, PU.1, and G-CSFR/M-CSFR/GM-CSFR, and activates differentiation to these lineages. To test whether single Flt3 downstream activated genes would be sufficient to cause the same effects, we transduced murine STAT3 and PU.1 cDNA into MEPs using retrovirus expression vectors, respectively. Both STAT3- and PU.1-transduced MEPs were capable to differentiate into IPCs and DCs in SCF and TPO supplemented media, and, with even higher efficiency, in SCF, TPO, and Flt3L supplemented cultures. While human Flt3-transduced MEPs maintained megakaryocyte-erythrocyte developmental potential and gained IPC, DC, and myelomonocytic cell potential, STAT3- and PU.1-transduced MEPs lost both megakaryocyte and erythrocyte developmental options, indicating that strong signaling of these transcription factors fully converted MEPs to the IPC, DC, and myelomonocytic lineages. Consistently, GATA-1 expression was down-regulated, and C/EBPα mRNA was up-regulated in STAT3- and PU.1-transduced MEPs. Interestingly, STAT3- and PU.1 over-expression in MEPs led to up-regulation of Flt3 mRNA levels, suggesting a self-sustaining effect of Flt3 signaling-cascade induced Flt3 expression. Thus, enforced expression of STAT3 and PU.1 in MEPs reprogrammed them to differentiate into IPCs, DCs and myelomonocytic cell lineages and inhibit Meg/E-lineage potential. Based on these data, we propose a model where Flt3 positive progenitor cells that locate in Flt3L rich environments will be directed to develop into IPCs and DCs. This process might be enhanced by a self-sustaining mechanism where Flt3 downstream transcription factors as STAT3 and PU.1 in turn maintain Flt3 expression.
Dose-Escalated Donor Lymphocyte Infusions Post Allogeneic Transplantation in Non-Hodgkin’s Lymphoma.
Authors: Bloor Adrian J.C. , Thomson Kirsty , Cooper Nichola , Peggs Karl S. , Mackinnon Stephen 
Published: October 2019

Abstract
The role of donor lymphocytes infusions (DLI) in the treatment of non-Hodgkin’s lymphoma (NHL) following allogeneic haemopoietic stem cell transplant (HCST) has not been clearly characterised. We report the results of 51 infusions of dose-escalating donor lymphocytes to 24 patients following myeloablative (n=3) or reduced intensity (n=21) transplantation as treatment for NHL. The diagnoses were low grade NHL (n=19), transformed low grade NHL and high grade NHL (n=5). Twenty patients were transplanted from an HLA-matched sibling, and 4 from an unrelated donor. The indications for DLI were progressive or relapsed disease (n=17), with or without mixed chimersim, and persistent mixed chimersim alone (n=7). Eight of the patients treated for disease progression or relapse had both clinical and radiological evidence of disease, a further 8 cases had radiological evidence of disease alone (CT scan in 3; CT-PET scan in 6) and one case of progressive disease was diagnosed following a bone marrow biopsy. Histological confirmation of relapse or disease progression was confirmed in 8 cases. Eighteen patients received an initial dose of 1x106/kg CD3 positive cells at a median of 265 days post transplant (range 181–728) and 6 received an initial dose of 1x107/kg cells at a median of 402 days post transplant (range 130–2674). Escalating doses of cells were administered at 3-month intervals (3 x 106/kg, 1 x 107/kg, 3 x 107/kg, 1 x 108/kg) in the absence of development of graft-versus-host disease (GvHD), if mixed chimerism persisted or if there was no evidence of disease response. Four of the patients treated for disease progression also received antitumoural chemotherapy shortly before DLI. Of the 16 evaluable patients treated for disease, 12 (75%) showed a significant response to DLI (9 patients with low grade NHL and 3 with transformed low grade NHL). A complete remission (CR) was observed in 10 patients and partial remission in 2 patients. Three of the patients achieving a CR following DLI subsequently relapsed; the median duration of the ongoing complete remissions is 574 days (range 122–1479). One patient died of sepsis before the response to DLI could be assessed. Conversion from mixed to multilineage full donor chimersim occurred in 18 of 18 (78%) evaluable patients. The major toxicity resulting from the use of DLI was GvHD. Following DLI, acute GvHD (grade II–IV) occurred in 5 out of 20 evaluable patients (24%) and chronic extensive GvHD in eight patients (38%); 1 patient died during treatment for grade IV acute gut GvHD. The incidence of GvHD did not correlate with the dose of DLI infused and 5 patients had graft-versus-lymphoma responses without GvHD. These data support the existence of a clinically significant graft versus tumour effect in non-Hodgkin’s lymphoma and indicate that this is an effective treatment for progressive disease post allogeneic HSCT.
ZAP-70 Status May Not Predict Outcome after Non-Myeloablative Allogeneic Transplantation (NMT) in Patients with Chronic Lymphocytic Leukemia(CLL) Who Failed Conventional Chemotherapy.
Authors: Khouri Issa F. , Saliba Rima M. , Keating Michael J. , Admirand Joan , O’Brien Susan , Champlin Richard E. , Buenos-Ramos Carlos 
Published: October 2019

Abstract
In aggressive disease, the CLL cells usually express the 70-kD zeta-associated protein (ZAP-70). We used immunohistochemical techniques and routinely fixed, paraffin-embedded tissue to survey ZAP-70 expression in patients (pts) with CLL who received a NMT, as previously described (Modem Pathol17:954, 2004). All cases were reviewed independently by two authors. Cytoplasmic staining of non-neoplastic, reactive T cells served as an internal control in each case. Neoplasms (Pax-5 +) demonstrating ZAP-70 cytoplasmic staining in greater than 20% of tumor cells were considered positive. Pts were eligible for NMT if they had failed fludarabine-based conventional therapy. The conditioning regimen prior to NMT consisted of fludarabine, cyclophosphamide and rituximab. Twenty nine pts were treated. The pts characteristics and outcome were as follows:
ZAP-70 Negative ZAP-70 Positive P value No. Pts 8 19 Age (range) 58 (45–69) yrs 53 (34–72) yrs 0.36 Time Dx to NMT 4.5 yrs 4.5 yrs 0.6 No. Prior Chemoregimens 3 (2–5) 3 (2–8) 0.7 No. Pts in Richter 1 6 0.3 Status at NMT CR/PR/NR 1/3/4 1/12/6 0.18 Donor: Matched Sibling 7 16 0.6 Unrelated 1 3 #Pts requiring DLI 3 9 0.4 Final Response CR/PR 6/1 15/0 0.6 NR/NE 1/0 3/1 Relapse Post CR 1 0 Follow-up Time 11 (9–47) mos 37 (13–75) mos Survival 1.5- year 83% 84% 0.7 4- year NA 64% Current PFS 56% (1.5 yr) 65% (4 yr) Causes of Death PD(1), cGVHD(1) PD(2), cGVHD(2), Infection(2) GVHD grade II–IV 37% 39%
These data suggest that NMT may overcome the negative prognostic impact of ZAP-positivity in CLL. Controlled trials are needed to confirm these results in a larger number of patients.
A Single Center Experience with Allogeneic Sibling Umbilical-Cord-Blood Transplantation in Children with Malignant and Non-Malignant Disease.
Authors: Song Yongping , Zhang Yanli , Fang Baijun , Wei Xudong , Lu Lulu , Zhu Xinhu , Wang Ping , Li Yufu , Gao Quanli , Wang Guiye , Qiu Lugui , Zhao Chunhua 
Published: October 2019

Abstract
To study the outcome of allogeneic sibling umbilical-cord-blood transplantation (CBT) in children in China, we studied the records of 38 recipients of cord blood from HLA-identical siblings from 1998 through 2004. The mean age at transplant was 10.3+/−5.5 years. Diseases of the patients were leukemia (28), neuroblastoma (3), severe aplastic anaemia (3) and inborn errors of metabolism (four). A volume of 76–208 ml of cord blood was collected from sibling infants soon after delivery. HLA antigens were identical in 32 and one to three antigens mismatched in six. The patients received busulphan and cyclophosphamide as conditioning, and antithymocyte globulin was given to prevent graft rejection. The median number of collected nucleated cells was 3.5 x 107/kg nucleated cells (range, 2.0 to 10.2). T-cell reconstitution was evaluated by using combined approaches of phenotyping analysis of lymphocytes and assessment of ex vivo thymic function by measuring T-cell receptor (TCR) rearrangement excision circles (TRECs). The overall survival is 89% at the median observation time of 72 months. The median time for neutrophil engraftment (absolute neutrophil count >500/mm3) was 16 days (range, 10 to 41days). The median time to become transfusion independent after CBT was 27 days for platelets (range, 15 to 54) and was 28 days for packed red blood cells (range, 19 to 128). Acute GVHD (aGVHD)was observed in 20/38 patients and involved only skin in 16 patients, skin and liver or gut in 2 patients and all 3 organs in another 2 patients. Seventeen of 20 patients had grade 1 to 2 aGVHD toxicity, whereas 3 patients experienced grade 3 to 4 aGVHD. Chronic GVHD (cGVHD) developed in 12 patients. Acute transplant related mortality was 5.5%. Cause of death was persisting non-engraftment till day +100 after transplant. Late mortality occurred in 2 patients: one cGVHD associated haemorrhage 22 months after CBT and one cGVHD associated septicaemia 4 years after CBT. Immunologic reconstitution demonstrated that CBT resulted in consistent and stable T-, B- and natural killer (NK) cell development. The kinetics of development was such that T-cell development occurred between 65 to 180 days. Initial T-cell engraftment consisted predominantly of CD45RO+, CD3+, and CD4+ T cells, and at 12 to 24 months changed to CD45RA+, CD3+, and CD4+ T cells, indicating de novo maturation of T cells. During the first 5 months after transplantation, TCR repertoires were highly abnormal and TREC values low. However, in a longer follow-up (one years or more after transplantation) TREC values and TCR diversity increased and became normal. This data confirms that CBT still should be the first treatment choice if an HLA-identical sibling is available.
Upfront Autologous Stem Cell Transplantation (SCT) Ameliorates the Prognostic Disadvantage of an Intermediate/High-Risk FLIPI Score in Patients with Advanced Follicular Lymphoma (FL): Evidence from Two Independent Data Sets.
Authors: Martin Kornacker , Suessmilch Simone , Gulla Hilde , Seyfarth Baerbel , Schoch Robert , Kneba Michael , Ho Anthony D. , Hensel Manfred , Schmitz Norbert , Dreger Peter 
Published: October 2019

Abstract
The Follicular Lymphoma International Prognostic Index (FLIPI) is a useful predictor for the prognosis of patients with FL. It relies on a score consisting of age, stage, hemoglobin, LDH, and number of nodal sites involved at diagnosis, which defines 3 risk groups with different 5-year survival (5-y OS): Low risk (0–1 factor; 5-y OS >90%); intermediate risk (2 factors; 5-y OS 78%); and high risk (>2 factors; 5-y OS 53%). We hypothesized that SCT in first remission might improve the course of patients with advanced FL and high/intermediate risk FLIPI, thereby abrogating the prognostic impact of the FLIPI.
Methods: Consecutive patients from two independent data sets (Kiel/Hamburg = training sample (T); Heidelberg = validation sample (V)) who received upfront SCT for FL between 1990 and 2002 and had sufficient baseline information available to assess the FLIPI were analyzed retrospectively. SCT eligibility criteria were similar for both series and comprised first remission of advanced FL, age <60, and performance status <2. Patients were treated with sequential therapy comprising 2–8 cycles of a CHOP-like regimen for remission induction, Dexa-BEAM (T) or HAM (V) for stem cell mobilization, and TBI/CY or BEAM for myeloablation prior to SCT.
Results: 49 (T) and 57 (V) patients were eligible. Time from diagnosis to transplant was 9 (5–21) and 14 (5–116) months, respectively. The FLIPI assigned 40 (82%) and 36 (63%) patients to the high/intermediate risk group. With a median follow-up from SCT of 64 (12–134) or 62 (1–132) months, respectively, outcome data were similar in both samples. 5-y OS from diagnosis was 94% (T) and 90% (V) for the high/intermediate risk group vs. 83% (T) and 87% (V) for the low-risk group (p=0.09 and p=0.4). 5-year progression-free survival (PFS) was 62% (T) and 77% (V) vs. 71% (T) and 77% (V) (p=0.35 and p=0.73). Combining all patients (n=106) allowed for examination of each prognostic group separately. Differences in 5-y OS from diagnosis (90% for high risk, 93% for intermediate risk and 85% for low risk) or in 5-y PFS (60% for high risk, 75% for intermediate risk and 78% for low risk) were not statistically significant.
Conclusions: Upfront SCT abrogates the prognostic impact of the FLIPI by improving the outcome of intermediate/high-risk patients but not of low-risk patients. Therefore the FLIPI might be a useful tool to select patients for prospective trials on SCT in FL who may particularly benefit from this procedure.
Factors Affecting Outcome of Patients with Refractory/Relapsed Hodgkin’s Disease (HD) Treated with IGEV Chemotherapy (CT) and High Dose Consolidation Therapy (HDT) with Stem Cell Rescue.
Authors: Balzarotti Monica , Spina Michele , Magagnoli Massimo , Chisesi Teodoro , Pinto Antonello , Castagna Luca , Demarco Monica , Michieli Mariagrazia , Nozza Andrea , Tirelli Umberto , Santoro Armando 
Published: October 2019

Abstract
Purpose: to identify prognostic factors in pts with relapsing/refractory HD treated with IGEV CT and HDT
Methods: From 01/98 to 01/05 104 patients (pts) from our institutions with relapsed/refractory HD received 4 induction cycles of IGEV (ifosfamide 2000 mg/mq d 1–4; gemcitabine 800 mg/mq d 1& 4; vinorelbine 20 mg/mq d 1; prednisolone 100 mg/mq d 1–4, and G-CSF) and HDT consolidation. Prognostic factors for freedom for progression (FFP) were analyzed by means of the chi-square and Fisher’s exact tests. Multivariate regression logistic analysis was then applied for variable associated with outcome.
Results: Main pt characteristics at accrual: M/F 56/48, median age 31 (range 17–65), refractory/relapsed following last CT course 40/64, median previous CT regimens: 1 (range 1–2), simptoms A/B 32/71, Hasenclever (IPS) score <3/≥ 4: 72/24 (8 pts not evaluable), LDH ratio ≤ 1/> 1: 68/35, previous radiotherapy (RT) yes/no: 62/42. After induction, 45 pts (43%) obtained complete remission (CR), 41(40%) partial remission (PR) and18 (17)% di not response (NR). Overall 89 pts received HDT and at the end of the treatment program 76 out of 104 pts were in CR. On an intent-to treat analysis, two-year freedom from progression (FFP) and overall survival (OS) for the whole series were 47% and 74,6%, respectively.
In univariate analysis FFP was negatively influenced by B symptoms, IPS score ≥ 4, LDH ratio > 1, refractory disease, and by response to IGEV (see table).
Multivariate analysis revealed that factors retaining statistical significance were refractory disease, B symptoms, high IPS score and chemoresistance (see table).
Conclusions: based on this analysis, a prognostic model for IGEV-treated patients with refractory/relapsed HD can be constructed.
Factors influencing FFP Factor ≥ FFP p (univariate) p(multivariate) Symptoms B/A 16/57% 0.003 0.007 IPS > 4/< 3 32/51% 0.0035 0.051 LDH >1/<1− 19/61% 0.003 n.s. refrac/relaps 22.5/62% < 0.001 0.012 < CR/CR to IGEV 32/68% 0.002 n.s. NR/CR + PR 23.5/52% 0.001 0.005