bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phosphotyrosine Levels by Flow Cytometric Assay for Therapeutic evaluation in Chronic Myelogenous Leukemias
Authors: Sun Xuemei , Chen Junhao , Jiang Pengjun , Zhu Xuejun 
Published: April 2020

Abstract
Assay of phosphotyrosine levels using flow cytometry has been used to identify patients with chronic myelogenous leukemia (CML) positive for the Bcr-Abl fusion gene. We hypothesized that clinical monitoring could identify treatment response through reductions in intra-granulocyte phosphotyrosine. Initially, we studied cell lines FDC-P1 and HL60 (Bcr-Abl–negative) and FDrv210 and K562 (Bcr-Abl–positive) with our technique. A fluorescein isothiocyanate-conjugated monoclonal antibody was used along with fluorescence-conjugated microspheres for reference (ratio of sample fluorescence: bead fluorescence=relative fluorescence unit [RFU]). Samples from 20 controls and 32 patients undergoing treatment were analyzed using the same method. Bcr-Abl status for each patient was confirmed using fluorescent in situ hybridization (FISH) or polymerase chain reaction gene amplification (PCR). Testing of cell lines consistently produced expected results. Patient values were found to be significantly higher than control values (P&lt;0.001) and values for patients with advanced disease were significantly higher than for patients with chronic-phase disease (P&lt; 0.05). Results of clinical monitoring were consistent with results from PCR. Two patients who received allogeneic stem cell transplantation had molecular remission confirmed by PCR and had a marked decrease in RFU value (from 62 to 5 and from 131 to 23). No such fluorescence change was observed in patients who achieved clinical remission. Flow cytometric analysis of phosphotyrosine levels is a reliable and convenient adjuvant technique for diagnosis of Bcr-Abl–positive leukemias and shows promise for serial evaluation of patients undergoing treatment.
Frequency of BCR/ABL Breakpoints in CML Venezuelan Patients : Contribution to a Possible Worldwide Distribution of BCR/ABL Transcripts Associated to Ethnicity
Authors: Carreño Fanny J. , Lopez Jose L , Mata Estluz K , Cardenas Leonor , Costa Osiris Da , Mendez Carlos , Utrera Rene 
Published: April 2020

Abstract
Chronic myeloid leukemia (CML) is a common clinical pathology characterized by the clone expansion of stem cells of myeloid linage. At the cytogenetic and molecular level CML is associated with one reciprocal and nonrandom translocation (t(9;22)(q34;q11)) which causes the BCR/ABL gene fusion in the 22nd chromosome (Philadelphia chromosome, pH+). This gene codifies for a chimerical protein functionally active and able to exert a transforming effect on all the cells carrying that gene. The transcripts b2a2 and b3a2 are the principal variants identified on the CML patients. Due to high frequency and variation in expression of b2a2 and b3a2 transcripts in CML patients, the researchers have tried to associate the expression of those transcripts with clinical and ethnic aspects. Nevertheless, the results have been contradictory and a direct correlation between transcripts BCR/ABL, the clinic and ethnicity has not been yet found. Although, geographic variation has been already revealed for others leukemia types, thus enhancing the accuracy of prognosis, until now a geographic distribution of CML transcripts remain unrevealed. We report here for the first time, the frequency of different BCR/ABL transcripts in 467 Venezuelan CML patients. The RT-PCR for those transcripts evidenced that 449 (96.1%) of patients had the gene fusion p210BCR/ABL, being 205 (45.6 %) b2a2, 187 (41.7%) b3a2 and 40 (8.8%) b2a2/b3a2 co-expression. The remaining 17 (3.9%) included the expression of variant P190BCR/ABL (e1a2) and its co-expressions: e1a2/b2a2, e1a2/b3a2 and e1a2/b2a2/b3a2. Most of the patients were between 21 and 60 years old and 53% were males. The analysis of automatic sequences showed the T/C polymorphism in some b2a2 transcripts, while b3a2 and co-expressions transcripts did not present it. These results were included in our study on the geographic distribution of the BCR/ABL transcripts. That study revealed that the Asian and Middle East countries (Malaysia, Thailand, Korea, India, China, Japan and Iran) showed the highest frequency of b3a2, while American countries (Brazil, Mexico and Venezuela) presented b2a2 as the highest frequency found. This geographical distribution of transcripts was found to have a pattern according to longitude: increasing the relative expression of b3a2 from west to east. As for the relative expression of b2a2 and b3a2 transcripts by ethnic groups, it showed three main groups: Asians and Middle Easterns express mainly b3a2 transcript, Europeans (Caucasians European) express both transcripts, whereas Mestizos (mulattos, Caucasians and Amerindians) express mainly b2a2 transcript. This geographical distribution pattern suggests that the relation between clinical features and CML transcripts could be masked by underlying factors such as ethnicity and geographic location.
Secondary Malignancies after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Neuburger Stefan , Hemmati Philipp , Terwey Theis , Massenkeil Gero , Dörken Bernd , Arnold Renate 
Published: April 2020

Abstract
Introduction: Allogeneic stem cell transplantation is a curative therapeutical option for patients (pts) with hematological diseases. As result of long term survival, late complications such as secondary malignancies are emerging.
Methods: Here we present a retrospective analysis of 589 pts (median age at transplantation 41 years, range 16–75, male 339, female 250) who underwent allogeneic stem cell transplantation in our institution between 1995 and 2007 (siblings n= 285, unrelated donors n= 304). Pts suffered from acute leukemia (n= 257), chronic myeloproliferative disorders (n= 120), myelodysplastic syndrome (n= 39), non-hodgkin-lymphomas (n= 41) and others (n= 132). Pts received conditioning with (n= 436) or without (n=152) 12 Gy total body irradiation (TBI).
Results: Up to 2007, 283 of 589 pts (48%) died of relapse or transplant related mortality (n=135 relapse, n= 84 infection, n= 36 graft-vs-host-disease (GVHD), n= 8 organ toxicity, n= 20 others or unknown). 305 out of 589 pts (52%) had survived with a median follow-up of 40 months (range 1–144 months). 21 out of 305 pts (6,9%) developed secondary malignancies at a mean of 5,1 years (range 1–10 years) after allogeneic stem cell transplantation. Localizations of secondary malignancies were skin (n= 11 basalioma, n= 2 melanoma), gut (n= 2 adenocarcinoma of the small intestine, n= 1 coloncarcinoma), oral cavity (n= 1 squamous cell cancer) and 3 lymphomas. 17/21 pts (81%) received a myeloablative conditioning with 12 Gy TBI, 4/21 pts (19%) a reduced intensity conditioning. Acute GVHD &gt; grade 2 and chronic GVHD extensive disease appeared in 48% and 67% respectively. Thus, these pts must be treated with prolonged immunosupressive therapy. 2/21 (9,5%) died of secondary malignancies (carcinoma of small intestine and colon) and one pts of accidential infection.
Conclusion: Long term surviver of allogeneic stem cell transplantation are at increased risk of a secondary malignancies. Most of the pts received TBI-based conditioning and suffered from chronic GvHD extensive disease with consecutive immunosupressive therapy. Life-long follow-up will be needed to detect secondary malignancies in early stage of disease which might offer curative therapeutical options. Therefore, in our institution all pts undergo annual skin screening program and are sensitized for possibility of secondary malignancies late after transplantation.
Peripheral Blood Stem Cell Mobilisation with Pegfilgrastim Versus Filgrastim in Children and Adolescents
Authors: Fritsch Peter , Schwinger Wolfgang , Lackner Herwig , Sovinz Petra , Benesch Martin , Urban Christian E. 
Published: April 2020

Abstract
Background: Several studies have shown that Pegfilgrastim, the long acting agent of rh-GCSF, is as effective as Filgrastim in children undergoing cytotoxic chemotherapy by reducing the duration of neutropenia without increased adverse events as bone pain and headache. Recent studies in adults have shown that Pegfilgrastim is also effective to mobilize CD 34+ stem cells. In contrast to Filgrastim studies showed a reduction of time to reach the peripheral blood CD 34+ cell peak, resulting in earlier performance of leukapheresis. Although the use of Pegfilgrastim in children after conventional chemotherapy is established, only few data have been published yet describing the ability to mobilize CD 34+ stem cells. The aim of the study was to compare the efficacy of Pegfilgrastim versus Filgrastim for CD 34+ stem cell mobilization in children and adolescents.
Methods: Patients admitted after 2006 were stimulated with Pegfilgrastim on day 4 following a chemotherapy course of the according treatment protocol followed by peripheral stem-cell-apheresis. Filgrastim was added when CD 34 + stem cells did not reach 15/μl between day 15 to 20. Patients were compared to historical controls (2002–2005) stimulated with conventional daily Filgrastim.
Results: 3 groups of patients were compared: Group 1 (historical group): 6 patients with Ewing’s Sarcoma undergoing chemotherapy according to the EURO-EWING protocol receiving Filgrastim 10–20 μg/kg (median 11 μg/kg) underwent a median of 3,8 peripheral stem cell collections (PSCC) between day 12–24 (median 15,4). A median of 16,9 CD 34+ stem cells/kg BW were collected. Group 2: 3 Patients with Ewing’s Sarcoma admitted after 2006 undergoing chemotherapy according to the EURO-EWING protocol received only Pegfilgrastim 100–200 μg/kg (median 166 μg/kg). 3 PSCC were performed between day 11–13 (median 12) and a median of 24,3 CD 34+ stem cells/kg BW were collected. Group 3: 5 Patients suffering from recurrent neoplasm [B-NHL (n=1), medulloblastoma (n=1), ependymoma (n=1) and germ cell tumor (n=2)]. Chemotherapy was performed according to relapse protocols followed by Pegfilgrastim 150–200μg/kg (median 188 μg/kg), however 2 patients needed further cytokine stimulation with Filgrastim 10–15μg/kg (median 12,5 μg/kg) combined with stem cell factor. A median of 3,2 PSCC was performed between day 8–30 (median 17). A median of 11,9 CD 34+ stem cells/kg BW were collected.
Discussion: Our data show that stem cell mobilisation with Pegfilgrastim seems to produce earlier CD34+ peaks and better CD34+ yields than Filgrastim in children when performed during primary or without previous long lasting chemotherapy. However patients with previous long lasting chemotherapy might need additional mobilisation therapy with Filgrastim and/or stem cell factor.
HHV6 and EBV Reactivations Associate with Encephalitis in Patients Post Allogeneic Haematopoietic Stem Cell Transplantation
Authors: Jaskula Emilia , Tarnowska Agnieszka , Suchnicki Krzysztof , Lange Andrzej 
Published: April 2020

Abstract
Herpes viruses reactivation is frequent post HSCT and constitutes a major complication when the reactivation becomes clinically symptomatic. 102 patients at several time points (1058 measurements for each Herpes virus, on average 10 measurements per one patient, from one week to one year post transplant) were investigated with the use of quantitative PCR for the presence of DNA viral copies in blood cells. CMV, EBV and HHV6 DNA copies in blood cells were determined with the use of the real-time PCR technique employing an Opticon 2 thermocycler (MJ Research, Watertown, USA). PCR conditions and the sequences of PCR primers and probes were selected from BALF5 region of EBV, US17 region of CMV and U67 region of HHV6 as described by Kimura et al, (1999), Machida et al, (2000) and Locatelli et al, (2000), respectively.
It was found: CMV and EBV DNA copies (above 100 copies/100 000 cells) were detected in 35% and 23% of patients, respectively. HHV6 reactivation was considered as significant at lower number of copies (above 10 copies/100 000 cells) and was found in 41% of HSCT recipients. Reactivations were more common at early stage post HSCT, with a peak between +31 and +60 day, then at later time. This difference was statistical significant for CMV (p=0.021) and similar trends were also seen when EBV and HHV6 reactivation were analyzed. CMV and EBV reactivations were associated with severe post transplant complications what resulted with poorer 5 year survival (30% vs 72% p=0.015 and 35% vs 65% p=0.016) in patients with CMV and EBV reactivation, respectively. This was not seen when HHV6 was considered. The poorest 5 year survival was seen in patients having aGvHD and presenting reactivation of CMV (23% vs 67%, p= 0.023). Patients with hepatitis were more frequently positive for CMV (0.54 vs 0.30, p=0.071) and EBV (0.41 vs 0.20, p=0.091) reactivation as compared to those lacking clinically apparent hepatitis. Full blown post transplant limbic encephalitis was seen in one case only and was associated with the presence of HHV6 DNA copies in blood (10 copies/100 000 cells) and in spinal fluid (5865 copies/200μl). However, in 11 cases some neurological, psychological and/or psychiatric symptoms were seen. Notably, among these encephalitis cases nine, six and three patients had HHV6, EBV and CMV reactivation, respectively. In five patients with encephalitis reactivation of two Herpes viruses were detected. Separate analysis of investigated Herpes viruses showed the statically significant association of encephalitis with HHV6 (0.75 vs 0.37 p=0.008) and EBV (0.54 vs 0.20 p=0.019) reactivation. Encephalitis was diagnosed in 25% and 21 percentages of pts with EBV and HHV6 reactive respectively. Seven out of 11 patients with encephalitis died. It appears that HHV6 and EBV reactivations may associate with encephalitis that affects the survival post HSCT.
Full Donor Engraftment without Chronic GvHD Following HLA-Matched Sibling Donor Haemopoietic Stem Cell Transplantation for Diamond Blackfan Anaemia
Authors: Alamelu Jayanthi , O’Boyle Farah , Killeen Natalie , New Helen , Atra Ayad , Dokal Inderjeet , Davis John , Foroni Letizia , Szydlo Richard M. , Roberts Irene , de la Fuente Josu 
Published: April 2020

Abstract
Long-term survival in Diamond Blackfan Anaemia (DBA) is compromised by transfusional iron overload and/or side effects of high-dose steroids as well as increasing recognition of the increased risks of aplasia and malignancy. Allogeneic haemopoietic stem cell transplantation (HSCT) can cure the haematological manifestations of DBA, but the optimal conditioning protocol is yet to be defined, particularly avoiding irradiation in view of the increased risk of malignancy. Four consecutive children with DBA were conditioned with oral busulfan 14 mg/kg (days-9 to -6), cyclophosphamide 200 mg/kg (days-5 to -2) and horse ALG 45 mg/kg (days-4 to -2) prior to HLA-matched sibling donor HSCT. Graft-versus-host disease (GvHD) prophylaxis was provided with ciclosporin from day-1 for six months to a target range of 100–150 ng/mL and methotrexate 10 mg/m2 on days +4 and +7.
P1: 10 years old boy who was transfusion dependent for two years having lost the response to steroids. In addition, he had deafness, short stature, gastrooesophageal reflux and cleft palate. He received bone marrow (8 × 108 TNC/kg). P2: 10 years old boy who was steroid dependent and had developed diffuse liver steatosis and bone fractures. In addition, he had short stature and undescended testis. He received bone marrow (1.91 × 108 TNC/kg). P3: four year old girl who was transfusion dependent with no associated abnormalities. She received a combination of cord blood (0.83 × 107 TNC/kg) and bone marrow (0.738 × 108 TNC/kg) from a sibling born following pre-implantation genetic diagnosis and HLA typing. P4: six year old boy born at 34 weeks of gestation who was transfusion dependent. He received a combination of cord blood (1.4 × 107 TNC/kg) and bone marrow (2.73 × 108 TNC/kg).
Neutrophil and platelet engraftment were prompt in all 4 patients (neutrophils: day 15–22; platelets day 10–33). Stable complete donor haemopoiesis was confirmed in all four patients using peripheral blood genomic DNA microsatellites markers by genescanning on day +28, 6 months and 1 year post-transplantation. The conditioning protocol was well tolerated by all patients with no life-threatening infections, though one patient had moderate veno-occlusive disease responding to defibrotide followed by gastrointestinal haemorrhage and another suffered reversible ciclosporin-associated posterior leucoencephalopathy and stage IV, steroid-responsive, acute gut GvHD. All four patients are alive with full donor haemopoiesis and normal haematological parameters at 11, 12, 15 and 24 months post-transplantation. Sibling allogeneic HSCT with this non-TBI protocol is a reasonable and feasible option for DBA patients suffering from unacceptable steroid side effects or chronic transfusion therapy.
Allogeneic Stem Cell Transplantation (SCT) in Advanced Chronic Myeloid Leukemia (CML) Patients after Tyrosine Kinase Inhibitor (TKI) Therapy
Authors: le Coutre Philipp D. , Hemmati Philipp , Neuburger Stefan , Terwey Theis , Massenkeil Gero , Daniel Peter , Dörken Bernd , Arnold Renate 
Published: April 2020

Abstract
In contrast to chronic phase CML patients (pts), in whom imatinib is the treatment of choice, in advanced CML pts allogeneic SCT remains the only therapeutic option with a curative potential. In order to adress the impact of pre-transplant parameters such as remission status, duration of disease, baseline therapies and cytogenetic and/or mutation status we analysed advanced CML pts with TKI pre-treatment (imatinib, nilotinib and/or dasatinib) who subsequently received an allogeneic stem cell transplant. A total of 24 pts (15 males, 9 females) was studied (median age 43,8 years, range: 21 – 58). Of these 7 pts were in 1. chronic phase (CP), 10 in 2nd or 3rd CP, 2 in accelerated phase (AP) and 5 in blast crisis (BC). Eleven of the 17 CP pts had either a CHR (complete hematologic remission) or HR at SCT and seven had a major cytogenetic remission (MCyR, including 4 CCyR). Cytogenetics and molecular analysis at SCT showed additional chromosomal abnormalities in 9/21 and point mutations in the BCR-ABL kinase domain in 2/17 pts (E255V & Y253H). All pts received imatinib prior to SCT, in 4 pts a 2nd line TKI therapy (dasatinib or nilotinib) and in one patient a 3rd line TKI therapy (dasatinib) was given. In 8/24 pts additional chemotherapy or irradiation after the last TKI were given prior to conditioning therapy. In 23/24 pts a myeloablative conditioning regimen was given. 15 pts were transplanted from an unrelated and 9 pts from an HLA identical family donor. The median follow-up of all patients is 19.8 months (range: 0 – 71). 12/24 pts died (SCT failure: n= 1, infection: n=5, GvHD: n=3, relapse: n=3). Risk stratification by basline parameters showed a statistically relevant survival advantage for patients with at least a MCyR and no non-TKI therapy prior to conditioning therapy (14 versus 10 pts; 14 versus 5 months; p=0.036; Chi-square test). Likewise, a survival benefit was identified for pts receiving SCT with at least a baseline HR (15 versus 9 pts; 11 versus 5 months; p=0.011; Chi-square test). Conversely, no statistically relevant differences were observed when pts were stratified upon additional chromosomal alterations, donor origin or stage of disease at baseline. In summary our data confirm the feasibility and benefit of allogeneic SCT in advanced CML patients following TKI treatment. Relevant pre SCT risk factors that could be identified as part of this analysis included baseline response and the absence of a non-TKI treatment prior to the conditioning therapy. Further studies are required to optimize the pretransplant treatment of advanced CML patients that are eligible for allo SCT.
Feasibility and Efficacy of BEAM as a Frontline High Dose Chemotherapy Supported by Autologous PBSCT in Elderly Patients (≥60 Years) with DLBCL
Authors: Ivanov Vadim , Coso Diane , Rey Jerome , Aurran Therese , Stoppa Anne-Marie , di Collela Jean-Marc Schiano , Blaise Didier , Bouabdallah Reda 
Published: April 2020

Abstract
Limited data is available concerning feasibility and efficacy of high dose therapy (HDT) supported by autologous PBSCT in elderly patients with non-Hodgkin lymphoma (NHL). In young patients with poor prognostic features intensification supported by PBSCT as a part of first-line treatment suggests survival benefit. It is not clear if the same strategy is applicable to the older patients. The Institute Paoli-Calmettes database was reviewed for all DLBCL patients who received BEAM followed by PBSCT in patients &gt;=60 years old between January 1998 and December 2006 (9 years). All patients were HIV-negative and received BEAM intensification as a part of front-line treatment. All of them were in a complete or partial response after CHOP or R-CHOP induction prior to autograft. Twenty seven auto-transplanted patients were identified (median age 63 y, range 60–68). This cohort was compared with closely matched group of 37 patients of same age range, who received first-line CHOP or R-CHOP regimen without intensification in the same 9-years interval. Only patients in a complete response after first line were included. As frontline autoPBSCT was performed in high-risk patients, the group without HDT was naturally privileged in the terms of Ann-Arbor stage and aaIPI index. There was significant difference in the localised vs disseminated disease (stage I–II: 54% in no-HDT vs 26% in HDT group, p=0.03)) and aaIPI (0–1: 66% in no-HDT vs 37% in HDT, p=0.046) between the two groups. Factors evaluated included treatment-related mortality (TRM), overall survival (OS) and event-free survival (EFS). TRM in the HDT group (1/27 pts (3,7%)) was comparable with previously published data. The estimated 5-year OS was 75,5% (95%CI 52–90 %) for HDT group compared to 79,9% (95%CI 58–92%) in the no-HDT group (p=0,75). There were 8 events (1 TRM and 7 relapses) in the HDT group and 11events (all relapses) in no-HDT (5-year EFS 49,4% vs 64,2%, p=0.45). We conclude that frontline autologous PBSCT with BEAM conditioning can be safely performed in patients aged 60 years or above with DLBCL after CHOP of R-CHOP induction. There was no difference in OS and EFS between cohorts with and without intensification even if the auto-transplantation procedure was reserved for the high risk patients. We conclude that first-line HDT with autologous PBSCT in older patients with high-risk IPI score might improve survival in this group and produce results similar to those in the low-risk group.