bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analytical Characterization of Infusion Volume Reduced Humate-P®/Haemate® P.
Authors: Metzner Hubert J. , Kanzy Ernst-Jürgen , Kalbass Antje , Lind Holger , Mueller Heinz-Georg , Kumpe Gerhard , Schulte Stefan 
Published: October 2019

Abstract
Commercially available von Willebrand factor (VWF)/factor VIII concentrates are therapy of choice in severe von Willebrand disease (VWD), clinically characterized by serious bleedings, as well as in some cases of hemophilia A. Humate-P®/Haemate® P is one of the few products approved for therapy and prophylaxis in both indications, VWD and hemophilia A. As it was desirable to further improve the convenience of application for the patients by reducing the application volume, an infusion volume reduced (i.v.r.) Humate-P®/Haemate® P was developed increasing the product concentration while keeping the well-known positive product characteristics. Eight i.v.r. Humate-P®/Haemate® P batches were produced and vials of each batch were reconstituted using the reduced volume of water for injection, i.e. 5 mL (250 IU F VIII), 10 mL (500 IU F VIII), for 15 mL (1.000 IU F VIII). These lots were compared analytically with 18 lots of current Humate-P®/Haemate® P. The analytical comparison demonstrated that the i.v.r. product contains twofold increased concentrations of the active ingredients VWF and F VIII (see table). As the purification and virus inactivation processes have not been changed, the quality of the active ingredients is maintained. This e.g. could be shown by a high VWF:RCo/F VIII:C ratio of about 2.4 for the volume reduced as well as the current product. Additionally, the VWF:RCo/VWF:Ag ratio and the high molecular weight (HMW) multimer structure of i.v.r. Humate-P®/Haemate® P are equivalent to current Humate-P®/Haemate® P (see table) and close to that of NHP indicating a comparably good efficacy of i.v.r. and current products. Based on the analytical investigations described here, it can be concluded that the development of i.v.r. Humate-P®/Haemate® P resulted in a product of unchanged quality of the active ingredients but very high VWF concentration. This allows the application of VWF in small volumes and, consequently, will furthermore improve the convenience of VWD and hemophilia A treatment.
Table Humate-P®/Haemate® P i.v.r. Humate-P®/Haemate® P Parameter 250/500 IU (n=12) 1000 IU (n=6) 250/500 IU (n=5) 1000 IU (n=3) F VIII:C (1-stage), IU/mL 24.5±1.3 31.2±2.0 51.2±0.8 66.3±4.2 VWF:RCo, IU/mL 58.3±4.1 78.3±4.0 119.8±10.9 159.3±14.2 VWF:RCo/VWF:Ag 0.78 ± 0.08 0.81 ± 0.06 VWF:RCo/F VIII:C (1-stage) 2.43 ± 0.12 2.36 ± 0.15 VWF HMW Multimers (>10) 75 ± 5.5% of NHP 77 ± 3.5% of NHP
Validation of TSE Agent Removal by Immunoaffinity and Ion Exchange Chromatography Steps of the ReFacto Manufacturing Process.
Authors: Booth James , Tannatt Molly , Vicik Steve , Kelley Brian 
Published: October 2019

Abstract
ReFacto® (moroctocog alfa), a recombinant Factor VIII approved for treatment of hemophilia A is produced by a mammalian cell culture process that includes therapeutic-grade human serum albumin (HSA) in the cell culture medium. The plasma-derived HSA used in the cell culture process is sourced from countries whose blood supply does not pose a significant transmissible spongiform encephalopathy (TSE) risk, and is manufactured utilizing a fractionation process that has demonstrated the capability to significantly reduce the level of experimentally added TSE infectivity. While to date, no case of TSE has been identified with a history of exposure to fractionated HSA, Wyeth has conducted a study to demonstrate that the ReFacto manufacturing process has significant capacity to remove a TSE agent if it were present in the HSA. The ReFacto purification process consists of five chromatographic steps, two of which were validated for removal of a TSE agent: the immunoaffinity step (8A4 Sepharose chromatography) and anion exchange step (Q Sepharose chromatography). The GLP study was performed using appropriately qualified lab-scale systems, which model the performance of the production equipment. Brain homogenates from scrapie-infected hamsters (263K strain) were added to loads of the respective chromatography steps and levels of the scrapie agent in the product pools were determined using a validated Western blot quantitation method. Duplicate runs were performed to assess the consistency of the TSE agent reduction. The results show that both chromatography steps provide significant clearance of the TSE agent, with greater than a billion-fold (9 log) reduction in infectivity provided by the two process steps in combination. The consistency of the duplicate run results was very good (less than 0.7 log difference between runs). While the risk of TSE contamination of HSA is only theoretical, these data provide a high degree of assurance that in the event of an adventitious TSE contamination of the HSA used in the ReFacto cell culture process, the purification steps would remove the infectious agent to safe levels.
Late Increase of Peripheral CD34+ Cells Number during Acute Liver Failure Recovery.
Authors: Drénou Bernard , Corlu Anne , Desille Yoann , Sicot Christèle , Fardel Olivier , Amiot Laurence , Lorho Richard , Camus Christophe , Guillygomarc’h Anne , Boudjema Karim , Guyader Dominique 
Published: October 2019

Abstract
Human Hematopoietic Stem Cells (HSC) of bone marrow origin are reported to transdifferentiate into various cell types, including cells of hepatic lineage, and to be involved in hepatic repopulation. As it has been previously reported in the course of myocardial infarction, we hypothesized that CD34+ cells would appear early in severe acute liver failure and could predict a favourable outcome.
Methods: We quantified circulating CD34+ cells in peripheral blood of 146 patients without and with severe hepatic failure (HF) defined by factor V <50%: 20 acute hepatitis with HF, 26 acute alcoholic hepatitis with HF, 22 major right hepatectomy (6/23 developped HF) and 78 controls (31 healthy subjects, 23 cirrhosis, 24 cirrhosis with chronic HF). CD34+ cells were quantified by flow cytometry, according to ISHAGE guidelines in both one and dual platform approaches. Phenotypic and functional analyses were performed in selected cases. In case of acute HF, quantification was done every day until correction of liver function. One patient underwent liver transplantation. The number of circulating CD34+ cells was correlated with factor V level. Results were presented using median and range.
Results : At inclusion, the numbers of CD34+ cells /mm3 were similar in different groups (healthy controls = 1.0 [0.1– 4.0]; cirrhosis = 0.9 [0.1–5.1]; cirrhosis with chronic HF = 1.0 [0.1–3.7]; alcoholic hepatitis with HF = 1.2 [0.1–6.0]; liver resection 24H after surgery = 0.8 [0.2–3.2]; acute hepatitis with HF = 0.7 [0.2–3.1]. For patients with acute hepatitis with HF, the number of CD34+ increased over time, from 0.7 [0.2–3.1] to 1.8 [0.5–9.6] at the 4th day after admission and was significantly (p <10–3) correlated with correction of factor V. There was no correlation between the number of CD34+ and the time between admission and onset of the disease. There was no significant increase over time in the hepatectomy group nor in the alcoholic hepatitis group. CD34+ cells detected during acute hepatitis regeneration presented an immature phenotype and coexpressed CD33, CD90, CD133, CD117, CD135 (FLT3-R), CD38, HLA-DR and not V-Cadherin. Clonogenic progenitor cell assays did not reveal specific features and correlation between number of CD34 and BFU-E colonies were in favour of their hematopoietic potentiality.
Conclusion: By contrast with data reported in acute myocardial infarction in which early mobilisation of CD34+ cells was observed, we found that the increase of circulating CD34+ HSC is a late event occurring at the time of the recovery of liver function in acute severe hepatic failure. The hematopoietic potentiality of mobilized CD34+ cells suggests that they do not play a major role in liver repopulation in fulminant acute hepatitis.
Mechanisms by Which NK T Cells Become the Predominant T Cell Subset in Mice after Irradiation.
Authors: Yao Zhenyu , Liu Yinping , McIntire Jennifer , Strober Samuel 
Published: October 2019

Abstract
Previously, we found that the percentage of NK T cells among all T cells in the spleen of mice treated with fractionated irradiation to the lymphoid tissues (Total lymphoid irradiation; TLI) with a total dose of 4,080 cGy increased markedly due to greater reduction in the absolute number of non-NK T cells as compared to NK T cells. The underlying mechanisms of the change in the T cell subsets after irradiation remained to be established. In the current study, C57BL/6 mice were given escalating single doses of 240, 1,000, 2,000 and 3,000 cGy total body irradiation (TBI). Splenocytes were harvested at 4 or 24 hours after irradiation, and the percentage and absolute number of NK T and non-NK T cells was determined. At the same time, the intracellular level of the anti-apoptotic protein, Bcl-2 was assayed by flow cytometry. In some studies, the turnover rate of NK T cells and non-NK T cells was examined by injection of BrdU and intracellular staining. At 4 hours after all doses of irradiation, neither the NK T nor non-NK T cell subset had a significant change in percentage or absolute number as compared to untreated controls. However, at 24 hours the percentage of NK T cells among all T cells had progressively increased with increased doses of TBI from 3% in the untreated controls to 65% in mice given 3,000 cGy. Whereas the absolute number of non-NK T cells decreased at least 1000 fold, the absolute number of NK T cells decreased approximately 50 fold after 3,000 cGy. The BrdU incorporation of NK T cells from irradiated mice was markedly reduced as compared to untreated mice, and was similar to that of non NK T cells in these irradiated mice. 8–12% of NK T cells and non NK T cells in untreated mice expressed a high level Bcl-2. As the dose of TBI increased progressively, the percentage of Bcl-2hi cells increased progressively to 89% amongst NK T cells and 70% amongst non-NK T cells. At each irradiation dose, the percentage of Bcl-2hi cells amongst NK T cells was higher than amongst non-NK T cells. There were 40×103 Bcl-2hi NK T cell and 10×103 Bcl-2hi non-NK T cells surviving per spleen at 24 hours after 3000 cGy TBI. The absolute number of Bcl-2hi NK T cells decreased by about two fold while the absolute number of Bcl-2hi non-NK T cells decreased by about 100 fold. These results indicate that the increased percentage of NK T cells amongst all T cells after irradiation is due to greater radioresistance rather than to more rapid replenishment of NK T cells as compared to non-NK T cells. We are investigating whether Bcl-2 plays a critical role in the extraordinary radioresistance of the NK T cells.
Peripheral Blood Progenitor Cell Apheresis Increases Circulating Stem Cells.
Authors: Chang Naibai , Pei Lei , Xu Xiaodong , Wei Jianping , Gu Xichun , Liu Hui 
Published: October 2019

Abstract
Peripheral blood hematopoietic stem cell transplantation has been replacing the bone marrow source stem cell transplantation for the treatment of hematological malignancies. Many efforts were made to enhance the stem cell number in peripheral blood and optimize the peripheral stem cell collection. Hematopoietic stem cell circles between bone marrow and peripheral blood during maturation. Thus the recruitment of hematopoietic stem cell in peripheral blood is a factor influencing the efficacy of peripheral blood stem cell apheresis. The aim of this study was to evaluate the effects of apheresis on the number of circulating stem cells. There were 41 consecutive cases involved. The mobilization regimen was chemotherapy + G-CSF. All patients underwent at least two apheresis. The volume of blood processed was calculated automatically by COBE Spectra cell separator. Blood samples were drawn just before each apheresis. CD34+ cells both in blood samples and in apheresis products were analyzed by multicolour flowcytometry. CFU-GM was cultured at same time. Data from the 1st and 2nd collection was compared. CD34+ /CD38− cells in peripheral blood increased from 48±12/ul to 133±40/ul (p<0.05) and CD34+/HLA-DR− cells from 45±12/ul to 97±29/ul (p<0.05) after apheresis. But CFU-GM was unchanged (154±35/105 cells to101±20/105 cells). CD34+ /CD38− cells, CD34+/HLA-DR− cells and CFU-GM in apheresis products were slightly increased but not reached the statistical significance(2.10±2.31x108 to 2.89±2.08x108, 2.09±2.35x108 to 2.32±1.61x108, 1.28±1.53x107 to1.93±3.11x107 respectively).
Conclusions: Peripheral blood stem cell apheresis promotes stem cell recruitment in peripheral blood and increases circulating hematopoietic stem cells.
Atypical Presentations of EBV Reactivation and EBV-Associated PTLD Post-HSCT: Implication for Routine EBV Surveillance after Allogeneic HSCT?.
Authors: Dobbelstein Christiane E. , Kamal Haytham M. , Dammann Elke , Hertenstein Bernd , Weissinger Eva M. , Ganser Arnold 
Published: October 2019

Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT), an effective treatment for leukemia and other disorders, produces profound immune deficiency in the early period after transplantation. Viral reactivations such as Epstein-Barr virus (EBV) are a major cause of morbidity and mortality after HSCT. EBV-associated posttransplant lymphoproliferative disease (PTLD) is a life-threating and often fatal complication occurring after allogeneic HSCT with a cumulative incidence of 1% at 10 years. Most cases occur during the first year after transplantation with the incidence highest at 2–3 months after HSCT. Typically PTLD presents as a disseminated disease with fever, lymph node enlargement, respiratory disease and feature of hepatitis. The common lymphadenopathy usually leads to diagnosis, with confirmation on viral tests. Here we retrospectively evaluate the patients at our institution since January 2002 to date (07/2005) who were allogeneic transplanted. Of the total 236 patients who were transplanted during this time period eight patients (3.4 %) developed a significant EBV reactivation with treatment need. Of the eight patients six had a fulminant course. Of these five patients died (EBV reactivation/PTLD is at least probable cause of death). Since at least two of the patients with the fulminant course presented atypically with fever as initial symptom and without other manifestations of PTLD like lymph node enlargement, these cases underline the need of an immediate pre-emptive therapy without any time delay for further diagnostic procedures e. g. diagnostic lymphadenectomy. While at present there is no recommendation of the EBMT for routine surveillance after HSCT, our data indicate the need for a standardized surveillance strategy.
UPN diagnosis conditioning GvHD prophylaxis HSCT max. viral load (EBV-DNA copies/ml) treatment outcome abbreviations: RIT: radioimmuno therapy; FLAMSA: fludarabine, amsacrine, cytosine arabinoside, cyclophosphamide, TBI 4Gy; TCD: T-cell depletion; (*): diagnosis post mortem. 888 AML Flu, Mel ATG, CsA, MTX, MMF MUD-PBSCT 11.0 x 10e6 (d +51) rituximab CR 908 AML TBI, Cy ATG, TCD MRD-PBSCT 1.75 x 10e6 (d +724) rituximab, foscavir, cyclophosphamide death 932 multiple myeloma Flu, Mel ATG, CsA, MTX MUD-PBSCT 2.3 x 10e6 (d +101) rituximab CR 973 AML RIT, TBI, Cy ATG, TCD MRD-PBSCT 10.0 x 10e6 (d +110) none (*) death 1140 multiple myeloma Flu, Mel ATG, CsA, MTX MUD-PBSCT 0.07 x 10e6 (d +278) rituximab death 1264 ALL TBI, Cy ATG, CsA, MTX MUD-PBSCT 1.75 x 10e6 (d +111) rituximab death 1271 AML FLAMSA ATG, CsA, MMF MMUD-BMT 7.0 x 10e6 (d +91) rituximab death 1300 aplastic anemia Flu, Cy, TBI ATG, CsA, MTX MRD-BMT 0.75x 10e6 (d +26) rituximab CR
Identifying Patients at Risk of Early Relapse after Submyeloablative Allogeneic HSCT.
Authors: Mehta Jayesh , Gordon L. , Tallman M. , Winter J. , Williams S. , Evens A. , Frankfurt O. , Grinblatt D. , Kaminer L. , Meagher R. , Singhal S. 
Published: October 2019

Abstract
Since graft-vs-tumor effects are the mainstay of sustained disease control after submyeloablative (mini) allogeneic transplantation, rapidly-progressive malignant disease may not be curable with SMAT. Figure 1 shows the cumulative incidence of chronic GVHD and relapse (n=35) in 63 patients undergoing mini-allografts for hematologic malignancies (acute leukemia, lymphoma/CLL, myeloma).
Figure Figure
The pattern suggests that a substantial number of relapses occur before chronic GVHD and associated GVT get established. The purpose of this analysis was to see if patients at risk of early relapse (relapse within 100 days; n=15 in this series) could be identified based on pre-transplant characteristics (diagnosis, chemosensitivity, Hb <9, platelets <100, abnormal LDH, albumin <3, performance status). In Cox analysis, the only factor predictive of early relapse was Hb <9 at the time of transplant (7 of 15 vs 8 of 48 with Hb ≥9; RR=3.1; P=0.029). The patients were also retrospectively classified by their physicians to have “slow” (n=33) or “rapid” (n=30) disease pre-transplant based on clinical impression. 11 early relapses occurred in patients with “rapid” disease and 4 in those with “slow” disease. When disease tempo was introduced in the Cox analysis as an additional variable, it was of borderline significance (P=0.051); with Hb <9 remaining as the sole predictive factor.
Figure Figure
The curves showing the temporal relationship between chronic GVHD and early relapse based upon Hb or disease tempo suggest that patients with Hb ≥9 or those with “slow” disease stand to benefit more than the others from graft-vs-tumor effects. Accordingly, restriciting mini-allografts to patients with Hb ≥9 or those with “slow” disease would be expected to improve outcome in terms of disease-free survival. However, neither of these two factors is particularly satisfactory: Hb <9 identifies only half the patients relapsing early, and the subjective assessment of disease tempo by the treating physician, while identifying a greater proportion of patients at risk of early relapse, excludes almost half the patients from the allograft. These data suggest that studies are needed in larger groups of patients to develop more robust (sensitive as well as specific) criteria to identify patients at risk of early relapse after mini-allografts. Since such patients are unlikely to benefit from a standard mini-allograft procedure, they may be better suited for alternatives such as conventional-intensity allografts, tandem autograft and mini-allograft procedures, or novel immunotherapeutic approaches applied very early after a mini-allograft.
Impact of High-Resolution HLA Matching on Outcomes of Unrelated Donor Hematopoietic Stem Cell Transplantation.
Authors: Kang Seok Yun , Kim Hyeoung Il , Lee Hyun Woo , Jang Jun Ho , Park Joon Seong , Choi Jin Hyuk , Lim Ho Yeong , Kim Hugh C. 
Published: October 2019

Abstract
Objective: We assessed the impact of high-resolution genotypic results of human leukocyte antigen (HLA) for all major class I and II loci between donors and recipients in the outcome of unrelated hematopoietic stem cell transplantation (HSCT).
Method: Between 1999 and 2005, high-resolution genotyping for HLA-A, -B, -C and -DRB1 was performed for 23 unrelated HSCT. All the patients were typed as HLA identical by serologic technique and then they were also typed HLA identical by high resolution technique. Unrelated bone marrow transplantation using DNA-based high resolution HLA compatibilities were considered in the analyses of clinical outcomes such as hematopoietic engraftment, acute GVHD, and survival. And then, we compared with patients who received related HSCT (same institute, same duration) and also unrelated HSCT data from IBMTR.
Results: Median follow up duration was 9 months (1–51). Fifteen patients were male and 8 were female. Median age was 22 years (range 6–52). Median time from diagnosis to transplantation was 7 months (range 4–63). Eight patients of acute myeloid leukemia (AML), 6 of chronic myeloid leukemia (CML, 2 of 6 were in blast crisis), 4 of acute lymphoid leukemia (ALL), 3 of severe aplastic anemia and each case of juvenile myelomonocytic leukemia and myelodysplastic syndrome were enrolled. Median value of total nucleated cell and CD34 positive cell count was 3.51 (1.06–20.7) x 108/kg and 4.88 (1.33–46.9) x 106/kg, respectively. The conditioning regimen and prophylaxis for graft versus host disease (GVHD) were not different from conventional HSCT except one case of non-myeloablative transplantation. Median value of granulocytic (absolute granulocyte count > 500/mm3) and platelet (> 20,000/mm3) engraftment were D + 16, D + 17, respectively. Grade II acute GVHD developed in 4 patients (2 patients subsequently proceded to chronic GVHD). Treatment related mortality was 2 out of 23 patients (8.7%). Median value of overall survival duration was 30 months. For AML patients, 3-year survival rate was 72.9 %.
Conclusions: Our survival data for unrelated HSCT based on high resolution genotyped HLA matching was superior to unrelated HSCT. Although the sample size is small, the survival data of AML patients (CR1 at transplant) was superior to the survivals of related HSCT, as well as that of unrelated HSCT. The findings were that transplantation using unrelated donors selected by high-resolution genotype identity improves the transplantation outcomes similar degree to the result of the related HSCT.