bluuebunny/crossref_metadata_2025_split huggingface dataset:
Simultaneous Transplantation of Three Cord Blood Units in Adults with High Risk Acute Leukemia.
Authors: Wiktor-Jedrzejczak Wieslaw , Rokicka Malgorzata , Urbanowska Elzbieta , Torosjan Tigran , Gronkowska Anna , Graczyk-Pol Elzbieta , Tomaszewska Agnieszka , Paluszewska Monika , Tormanowska Magdalena , Krol Malgorzata , Krol Maria , Jolkowska Justyna , Witt Michal 
Published: October 2019

Abstract
Adult patients with high risk acute leukemia have allogeneic hematopoietic transplantation as the only therapeutic option with some curative potential. Those who lack related or unrelated marrow donor are deprived of this possibility. Patients of smaller size may benefit from the transplantation of single cord blood unit, but for heavier patients two units may be required as reported by Barker et al. (Blood2005;105:1343). For very heavy patients even more units would be necessary to approximate the preferred threshold of 37 millions of cells per kilo. Moreover, cord blood units have variable size and it is always a risk that the only HLA-matched units for given patients would be small and two units would not provide adequate number of cells even for patients of moderate size. This is yet another argument to attempt transplantation of more than two units. Three patients have so far been included. Patient No 1 was 24 yrs old male (body weight 87 kg) with resistant AML transplanted with 3 units (two matched at 4/6 HLA antigens, one matched at 3/6 HLA antigens) on March 29, 2004 following conditioning with standard BuCy4 + ATG regimen. This patient died d. +42 of Gram negative sepsis without hematopoietic reconstitution. Patient No 2 is 22 yrs old male (body weight 100 kg) with Ph+ ALL in complete remission, who was transplanted on October 11, 2004 with 3 units (two matched at 4/6 HLA antigens, one matched at 3/6 antigens) following conditioning with TBI, etoposide and ATG. This patient had neutrophil recovery d. +27, and platelet recovery d. +49. He is alive and well 9 months after the transplant fully chimeric with progeny of only one of transplanted units. Patient No 3 is 31 yrs old male (body weight of 100 kg) with ALL relapsed after autologous BMT with multiple cytogenetic abnormalities. He was transplanted May 24, 2005 with 3 units (one matched at 5/6 HLA antigens, two matched at 4/6 HLA antigens) following conditioning with treosulfan, cyclophosphamide, melphalan and ATG (he earlier received TBI prior auto-BMT). Patient obtained neutrophil recovery d. +36 and as of d.+60 still has low platelets. No data on chimerism are yet available. These data confirm that transplantation of more than two cord blood units is feasible and may provide therapeutic option for patients with high risk leukemia who lack marrow donor and for whom two cord blood units do not provide sufficient number of cells. As in the case of the transplantation of two units patients seem to be reconstituted with progeny of only one “winning” unit.
HIV-1 Vector Mediated Expression of Human B-Domain Deleted FVIII in Primary Canine Marrow Mononuclear Cells.
Authors: Schuening Friedrich G. , Srinivasakumar Narasimhachar , Zaboikin Michail M. , Zaboikina Tatiana N. 
Published: October 2019

Abstract
We wished to determine if primary canine marrow mononuclear cells (MNC) would support the expression of human FVIII and thereby serve as a potential target for gene therapy of hemophilia A. To this end, we created HIV-1 vectors that express B-domain deleted human FVIII (BDFVIII) using the viral LTR promoter or constitutively active cellular promoters: the mouse phosphoglycerate kinase (PGK) or elongation factor 1α (EF1α) promoters. The vector that expressed BDFVIII from the LTR also encoded a functional single-exon Tat protein [Srinivasakumar and Schuening (2000) J. Virol.74:6659–6668]. These promoters have been previously shown, by us and others, to express transgenes at high levels in hematopoietic cells [Srinivasakumar et. al., (2002). J. Virol.76:7334–7342]. Separate vector stocks were prepared for each of the above vectors by transient transfection of 293T cells [Srinivasakumar (2002) Methods Mol. Med. 69:275–302] and their titers were determined by real-time quantitative PCR of genomic DNA isolated from vector transduced 293T cells (Table 1). Each vector stock was then used for infection of primary canine marrow MNC by three spin-infections over 2 days on CH296-coated 6-well plates. The medium was replaced at the end of the transduction procedure. Conditioned medium was harvested 3-days after transduction and assayed for functional FVIII activity using the Coamatic Factor VIII kit (Chromogenix, Monza, Italy). The results, shown in Table 1, indicate that vector-derived FVIII could be detected in the supernatants of canine marrow MNCs transduced with all HIV-1 vectors. Highest levels of functional FVIII were obtained with the Tat-encoding bicistronic HIV-1 vector. These results demonstrate that canine marrow MNC support expression and secretion of functional human FVIII. Further experiments are warranted to determine if the levels of FVIII observed in vitro will translate into therapeutic benefit in vivo in canine models of hemophilia A.
Table 1. Expression of human BDFVIII in canine marrow mononuclear cells transduced by HIV-1 vectors.
High-Throughput DNA Analysis for Red Blood Cell Antigens Facilitates the Transfusion Support with Antigen-Matched Blood in Sickle Cell Disease Patients.
Authors: Castilho Lilian , Hashmi Ghazala , Reid Marion E. , Shariff Tasmia , Seul Michael , Pellegrino Jordao , Costa Fernando F. 
Published: October 2019

Abstract
Background: Transfusion dependent patients as those with Sickle Cell Disease (SCD) patients become alloimmunized and have the potential to form additional antibodies with such frequency that antigen-negative blood is preferred to prevent further alloimmunization. Blood group genotyping is playing a supporting role in the routine blood banks, especially for provision of antigen-matched blood for these patients. However, current techniques for genomic typing are all labor-intensive and require manual set up and analysis by gel electrophoresis. As a result, DNA microarrays are being developed for the single nucleotide polymorphisms (SNPs) detection in the blood group genes to provide a fast procedure and an automated analysis of numerous blood group polymorphisms. We evaluated the usefulness of DNA microarray to provide a means to precisely match donor blood to the antigen-negative type of SCD patients. Method: A total of 12 DNA samples from patients with SCD (homozygous for HbS) and 84 DNA samples from blood donors, were analyzed by the HEA Beadchip (Hashmi et al, 2005) containing a total of 18 SNPs (FYA/B, FY-GATA, FY265, DOA/B (nt 378, 624, 793), COA/B, LWA/B, DIA/B, SC1/SC2, M/N, S/s, LUA/B, KEL1/2, JKA/B, DO323, DO350, HgbS) in a single reaction. Results: A genotype result was obtained for all SNPs tested on 96 samples within 4 hours of the start of testing. Results obtained by Beadchip analysis in donors were used to provide antigen-matched blood for FYA/B, FY-GATA, FY265, DOA/B, M/N, S/s KEL1/2, JKA/B, for all 12 SCD patients. This technology provided a fast procedure and facilitated the transfusion support with antigen-matched blood in SCD patients allowing the reduction of alloimunization to blood group antigens. Conclusion: This high-throughput DNA analysis has the potential not only to increase the inventory of antigen-negative blood but also to facilitate the matching of RBC component to the recipient’s blood type. It also contributes to the management of transfusions in SCD patients by allowing a more accurate selection of donor units. The application of microarray technology in transfusion medicine may have a tremendous impact on further improvement of the safety of blood transfusion.
A New Methodology To Screen Hemoglobinopathies Using Capillary Blood Samples.
Authors: Bento Celeste , Relvas Luis , Rebelo Umbelina , Vazão Helena , Campos Joana , Ribeiro Leticia 
Published: October 2019

Abstract
The prevalence of β-thalassemia carriers among the Portuguese was around 2–3%, but nowadays, due to the immigrant populations coming from Africa, Brazil, East Europe and Asia, with different forms of thalassemias and Hb variants, hemoglobinopathies are much more common and with different genetic characteristics. Concerned with the risk of having an increasing number of patients with the severe forms of hemoglobinopathies, we decided to screen carriers, after personal informed consent, among pregnant women until 18-weeks of gestation and young adults attending primary care services. The goal of the project is to screen approximately 50000 blood samples in order to:
know the hemoglobinopathies frequency and heterogeneity in the population living in Central Portugal (~2500000 inhabitants), and, based on these data, to develop a future cost/effective strategy for carrier identification; identify carriers to provide genetic information and counseling.
In primary care medical centers there are no facilities to collect venous blood samples and we needed to establish a methodology to identify Hb variants and to correctly quantify HbA2 and Hb F in capillary blood samples, which has to be send by ordinary mail to our Lab Center. Working with the “HbA1c Capillary Collection System” from BioRad, we settled an accurate procedure to perform HPLC analysis on capillary blood stored at room temperature until 7 days after collection. To validate the technique we tested, in diverse conditions, more than 200 random and known controls samples: for the same individuals, HPLC results in capillary blood, collected with this system, and in EDTA peripheral blood samples, were identical. This methodology will detect β- and δβ-thalassemias and Hb variants; molecular characterization can be done through blood spots in filter paper (Guthrie spots) collected at the same time. Within 24–48 hours after sample reception in the Lab, results are reported to the respective physician, who is going to establish the correlation with the hematological parameters.
Carriers identified through the screening will be urged to have their partner tested. If they are both carriers, they will be sent to our out patient clinic in order to evaluate the risk of having a child with a clinically significant hemoglobinopathy. If indicated, underling mutation will be identified, genetic counseling provided and prenatal diagnosis offered.
The project, supported by the national program Saúde XXI/FEDER/FSE, started last April, was preceded by training sessions for doctors and nurses working in primary care centers and maternity hospitals. Brochures and posters are distributed for public information. As reported in some European surveys, we are willing to be effective in preventing the birth of affected infants with sickle cell disease and severe forms of thalassemia in Portugal.
8 and 11Gauge Bone Marrow Biopsy Needle Performance Characteristics.
Authors: Goldenberg Alec , Kelley Priscilla , Ibrahim Sherif , Sen Filiz , Liu Cynthia 
Published: October 2019

Abstract
Introduction. An ideal biopsy needle consistently recovers representative specimens of adequate length with minimal dependence on patient characteristics. We have shown that bone marrow biopsy needle gauge influences the length and quantity of recovered specimens. A needle that performs independently of patient characteristics should yield specimens demonstrating a normal distribution of specimen lengths. We evaluated the influence of age and needle gauge on the specimen length distribution curves of patients undergoing bone marrow biopsies using SNARECOIL needles.
Methods. 88 bone marrow core specimens were recovered from 72 patients using 11G SNARECOIL bone marrow biopsy needles. The mean age of the patients was 61.4 and the m/f ratio was 45/27. 53 specimens were recovered from 39 patients (mean age = 47.1, m/f = 27/12) ≤ 64 and 35 specimens were recovered from 33 patients (mean age = 78.6, m/f = 18/15) ≥ 65. 106 patients underwent 127 bone marrow biopsy procedures using 8G SNARECOIL needles. The m/f ratio was 56/50 and the mean age of the patients was 63.1 years. 72 specimens were recovered from 56 patients (mean age = 51.4, m/f = 30/26) ≤ 64 years old and 55 specimens were recovered from 49 patients (mean age = 76.1, m/f = 26/23) ≥ 65. 40 additional specimens were prospectively recovered from 39 patients ≥ 65 (mean age = 74.3, m/f = 19/20) with 8G SNARECOIL needles.
Results. The mean specimen lengths of the 11G and 8G specimens were statistically the same (mean±SEM, 1.97±0.07 cm vs. 1.99±0.05 cm, respectively, p = 0.8). However the 11G specimen length frequency distribution curve deviated markedly from a normal distribution (skewness(skw) = 0.52) while the 8G specimen distribution was nearly normal (skw = 0.04). While the frequency distribution curves of older patients (≥ 65) biopsied with 8G needles showed minimal deviation from normality (skw = 0.12,) the distribution curve of older patients (≥65) biopsied with 11G needles deviated markedly from a normal curve (skw = 0.64). 40 specimens prospectively obtained from patients ≥ 65 demonstrated a mean specimen length of 1.77±0.09 cm with a near normal specimen length frequency distribution curve (skw = 0.07).
Conclusions. 1. Although commonly reported, mean specimen lengths do not completely characterize the performance of biopsy needles. 2.0 Specimen length frequency distribution curves provide added characterization of needle performance in defined patient cohorts. 3.0 In the older patient population, near-normal 8G specimen frequency distributions and skewed 11G distributions suggest that 8G specimens may provide more representative sampling.
A Randomized Phase III Trial of Low Dose Liposomal Amphotericin B (L-AmB) as Prophylaxis of Invasive Fungal Infections (IFI) in Neutropenic Patients (pts).
Authors: Penack Olaf , Schwartz Stefan , Martus Peter , Reinwald Mark , Schmidt-Hieber Martin , Thiel Eckhard , Blau Igor W. 
Published: October 2019

Abstract
Background The prophylactic use of systemic antifungal agents reduces morbidity and mortality after allogeneic hemopoietic stem cell transplantation (HSCT). However, the efficacy of this approach in pts receiving intensive chemotherapy or autologous HSCT is yet unproven. This trial was designed to evaluate the efficacy of L-AmB prophylaxis in high-risk neutropenic pts.
Methods: 231 pts with hematological malignancies and expected neutropenia (N) of more than 10 days (d) following intensive chemotherapy or autologous HSCT were enrolled, 219 pts became neutropenic and were randomized to receive either 50 mg L-AmB i.v. every second d (arm A) or no systemic antifungal prophylaxis (arm B). Treatment with L-AmB started 1–3 d prior onset of N and was continued until neutrophil recovery, breakthrough IFI, intolerable toxicity or death. The level of significance was 0.05 (two-sided) for all statistical analyses. Calculations were performed using commercially (SPSSWIN 12.0) software, the calculations for GEE were performed using the software MAREG.
Results Pt. characteristics: Eligible pts 219; arm A: 110; arm B: 109. Reasons for exclusion were: Absence of N (8), infection prior N (3) and pts decision (1). Baseline characteristics were balanced for age (mean 53.8 years), underlying disease (119 AML, 27 ALL, 64 NHL, 9 other), duration of N (mean 14.8 D) and treatment modality (primary 149, secondary 42, transplant 28). Primary endpoint: The incidence of proven and probable IFI was 5 of 110 pts (4.6%) in arm A and 22 of 109 pts (20.2%) in arm B (p = 0.001, RR = 2.9, CI 1.3 – 6.5). Key secondary endpoints: Pneumonia of unknown origin occurred in 6 pts (5.5%) vs. 28 pts (25.7%) (p < 0.001), the incidence of possible, probable and proven IFI was 11 pts (10.2%) vs. 42 pts (39.6%) (p < 0.001), systemic antifungals were used in 24 pts (22%) vs. 64 pts (59%) (p < 0.001), and death occurred in 4 pts (3.7%) vs. 9 pts (8.2%) (p = 0.16) in arm A vs. arm B. Toxicity: No grade 3 or 4 toxicity was noted. Laboratory abnormalities, including creatinine and liver function tests, were not different between the treatment groups.
Conclusion: Intermittent application of low dose L-AmB is save. The significant lower incidence of IFI in pts treated with L-AmB prophylaxis supports its use in prolonged N.
Prognostic Significance of Minimal Residual Disease (MRD) in Children with High Risk Acute Lymphoblastic Leukemia(ALL) A Children’s Oncology Group Study.
Authors: Borowitz Michael J. , Devidas Meenakshi , Bowman W. Paul , Larsen Eric , Pullen Jeanette , Camitta Bruce 
Published: October 2019

Abstract
MRD is known to be an important prognostic factor in childhood ALL. In POG 9906 we studied a select group of 271 patients with NCI high risk precursor B-ALL further defined by an age/sex/WBC algorithm (Shuster et al. Cancer Res Therapy Control9:101,1999) to select those at highest risk; historical data suggested that patients selected by this algorithm would account for about 12% of patients and have a 44% 4y EFS. Patients with Ph+ ALL or favorable genetics (TEL-AML1 translocation or trisomies 4 and 10) were excluded. All patients received 4-drug induction with vincristine, prednisone, L-asparaginase and daunomycin, plus intrathecal methotrexate, and consolidation with cyclophosphamide, ara-C and 6-MP followed by 2 cycles of interim maintenance and delayed intensification similar but not identical to so-called “augmented BFM”. MRD was measured in peripheral blood on day 8 (PB) and in bone marrow at day 29 (BM) by 4-color flow cytometry as previously described (Leukemia17:1566,2003). BM MRD data were available on 240 patients, and PB data on 243. At cutoffs of 1%, 0.1% and .01%, MRD in BM was associated with increased relapse rate (p=.0002). At a cutoff of .01%, 83/240 (35%) patients were MRD positive; these had 3 y EFS of 58±6% compared to 79±5% for MRD negative patients (p=.0008). The EFS of the 28 patients with MRD >1% was 44±14%. There were numerous (25/52) extramedullary events, mostly in CNS. MRD predicted 17/27 marrow relapses but only 8/25 other relapses (p=.03). Overall, 21% of MRD positive patients (>.01%) had a marrow relapse compared to 6% of MRD negative ones. Day 8 PB MRD was also prognostic, with the 49 MRD negative patients having a 3 y EFS of 89±6% compared to 70±5% for the 194 patients with MRD >.01% (p=.008). End induction BM MRD is a strong prognostic factor even among patients with especially high risk ALL and is a better predictor of marrow than of extramedullary relapse. Absence of MRD in day 8 PB identifies a subgroup of approximately 20% of these high risk patients who have an exceptionally good EFS, particularly given their very poor expected outcome.
EFS by Day 28 Marrow MRD EFS by Day 28 Marrow MRD
Toxicity of the Oncoprotein LMP1 of Epstein Barr Virus as a New Mechanism of LMP1 Regulation Explaining the Heterogeneity of LMP1 Protein Levels in EBV-Transformed B-Cells.
Authors: Christophe Le Clorennec , Tan-Sothea Ouk , Chantal Jayat-Vignoles , Ibtissam Youlyouz-Marfak , Eric Adriaenssens , Jean Coll , Dominique Bordessoule , Jean Feuillard 
Published: October 2019

Abstract
Epstein Barr Virus (EBV) oncoprotein LMP1 has been described as the major transforming protein in various cell types, by rerouting the TNF-receptor family signalling pathway. Despite this undoubtedly role in EBV-associated transformation of cells, toxicity of LMP1 is a well known experimental feature, poorly studied, may be because it contradicts the dogma of its transformation power. This phenomenon may be important to understand in the view of natural immune control of the EBV burden in the immunocompetent host (Le Clorennec, 2006). Moreover it is noteworthy that, at the single cell level, expression of LMP1 is very heterogeneous both in EBV-associated tumours and in cell lines.
To understand the toxic effect of LMP1 in LCLs, we cloned wild type LMP1 into a novel double inducible episomal vector pRT-1, from which the gene of interest is expressed under the control of a bi-directional doxycyclin regulatable promoter with a truncated version of NGFR (NGFRt) as a surrogate marker of inductibility.
We first found that increasing levels of doxycyclin concentration induce over-expression of CD95 that parallels induction of LMP1. Induction of LMP1 resulted in induction of CD95 aggregation, FADD phosphorylation, activation of caspase 8, 9 and 3, followed by late stages of apoptosis as demonstrated by PARP cleavage and sub-G1 peak. Immunoprecipitation of CD95 and caspase 8 suggested that LMP1 induction was associated with formation of CD95-dependant DISC.
Induction oxygen species (OS) has been shown to induce aggregation and autoactivation of CD95. We found that inhibition of the formation of oxygen species (OS) by catalase resulted in a decrease of LMP1-dependant apoptosis, whereas treatment of cells with H2O2 had the opposite effect. We also show that induction of latency III is associated with induction of OS. Levels of OS in latency III EBV-infected B-cells were significantly increased by induction of LMP1.
Therefore, our results are in agreement with a model according which (i) EBV latency III increases the pool of OS of the infected B-cell, rendering the cell permissive to CD95 auto-activation, (ii) increasing levels of LMP1 induces both expression of the pro-apoptotic CD95 and survival of the cell through NF-kappa B activation, and (iii) increasing CD95-autoactivation due to inappropriate over-expression of LMP1 finally overwhelms the antiapoptotic effect of NF-kappa B.
From this model, it is expected that at the single cell level, LMP1 expression results from an equilibrium between NF-kappa B dependant survival of cells and CD95-dependant apoptosis, explaining the heterogeneity of LMP1 expression from cell to cell and pointing a new mechanism of regulation of LMP1.
Ubiquitin-Conjugating Enzyme Ubc13 Is a Critical Component of TRAF-Mediated Inflammatory Responses.
Authors: Fukushima Toru , Matsuzawa Shu-ichi , Kress Christinia L. , Bruey Jean Marie , Krajewska Maryla , Lefebvre Sophie , Zapata Juan M. , Ronai Ze’ev , Reed John C. 
Published: October 2019

Abstract
Ubc13 is an ubiquitin-conjugating enzyme responsible for non-canonical ubiquitination of TRAF-family adapter proteins involved in Toll-like Receptor (TLR) and TNF-family cytokine receptor signaling. Gene ablation was used to study the function of Ubc13 in mice. While homozygous ubc13 gene disruption resulted in embryonic lethality, heterozygous ubc13+/− mice appeared normal without alterations in immune cell populations. Haploinsufficient ubc13+/− mice were resistant to lipopolysaccharide (LPS)-induced lethality, and demonstrated reduced in vivo ubiquitination of TRAF6. Macrophages and splenocytes isolated from ubc13+/− mice exhibited reduced LPS-inducible cytokine secretion and impaired activation of TRAF-dependent signal transduction pathways (NF-kB, JNK, p38 MAPK). These findings document a critical role for Ubc13 in inflammatory responses, and suggest that agents reducing Ubc13 activity could have therapeutic utility.