bluuebunny/crossref_metadata_2025_split huggingface dataset:
Control of Cell Death Levels Using Th9402-Based PDT Treatment on Fresh PBMC, and Potential Application to Patients with cGvHD.
Authors: Levesque Annie , Savard Ann-Louise , Roy Denis-Claude , Foss Francine , Scotto Christian 
Published: April 2020

Abstract
Recently, extracorporeal photopheresis (ECP) has shown interesting clinical activity for the treatment of drug-refractory chronic graft-vs-host disease (cGvHD), inducing Th1/Th2 immunomodulation to restore immune tolerance. Several studies indicate that target cell apoptosis may have a role in the control of cGVHD, and increasing apoptotic levels may favor immune modulation. We have developed an approach to eliminate immunoreactive cells using the Theralux™ photodynamic cell therapy (PDT) system based on the use of the rhodamine-derivative TH9402 illuminated ex vivo with a visible light source (l=514nm). The capacity of TH9402 PDT to induce increasing levels of T cell apoptosis has not been investigated. The induction of apoptotic cell death was studied using peripheral blood mononuclear cells (PBMCs) isolated from healthy volunteers (HV) and cGvHD patients. We found a good correlation between PDT conditions and levels of cell death induced. In comparison to controls showing 8±4 percent (%) of apoptosis, as measured by TUNEL assay on cells harvested 3 days post-treatment, PBMC from HV subjected to PDT using 0.33, 0.66, or 1.32 micromolar of TH9402 showed 28±16%, 49±17%, and 78±11% of apoptosis, respectively. These studies have also shown that the intra- and inter-donor variability in TH9402 incorporation are very low (~5% and 10%, respectively). To ensure that these findings could also be applied to a clinical setting, PBMC from cGvHD patients were treated with 0.33 and 1.32 micromolar of TH9402 to trigger either low or high levels of apoptosis. PBMC from cGvHD patients showed a sensitivity similar to that of PBMC from HV, with 38±9% and 73±13% of apoptosis when treated with 0.33 and 1.32 micromolar TH9402, respectively. Data obtained from the analysis of various cell death parameters such as the loss of DΨm, AnV/PI staining, TUNEL assay and agarose DNA ladder on PBMC and human Jurkat T cells treated with escalating doses of TH9402 suggested strongly the higher sensitivity of proliferating cells to the treatment. Moreover, together with the use of the caspase-inhibitor ZVAD-fmk, these data suggested that upon irradiation, the photoactivation of TH9402 will trigger the formation of reactive oxygen species (ROS) and the release of proapoptotic factors from mitochondria triggering various cell death mechanisms, such as caspase-dependent apoptosis, caspase-independent apoptosis, or a mixture of apoptosis and necrosis. Finally, preliminary data showed that PDT-treated cells were able to induce in vitro the maturation of monocyte-derived dendritic cells. Based on these data, we are beginning a pilot clinical study evaluating two controlled PDT conditions inducing low and high levels of apoptosis in order to assess the efficacy and biological effect of TH9402-based ECP to treat cGVHD in humans.
Selection of CD34+ HSC from Pooled Cord Blood Samples Does Not Result in Co-Purification of Potential Alloreactive CD3+ T Cells.
Authors: Gilmore Gary L. , DePasquale Darlene K. , Lister John , Shadduck Richard K. 
Published: April 2020

Abstract
Most systems for studying the ex vivo expansion of human UCB-HSC require enrichment of the HSC population, typically by CD34 selection. Because the number of HSC present in a single UCB collection is limited, one approach to increase the total HSC is to pool multiple UCB samples. A potential complicating factor with this method is that alloreactive T lymphocytes present in whole UCB samples would be expected to bind to cells displaying allogeneic HLA, including HSC, and would therefore co-purify with CD34+ UCB-HSC during isolation. Such complexes would be excluded from ISHAGE analysis by the side scatter gate, which is set to exclude aggregates. Alloreactive T cells would be expected to contain cytotoxic T lymphocytes [CTL], which could potentially inhibit or completely abrogate HSC expansion.
In order to determine whether CD3+ T lymphocytes co-purify with CD34+ UCB-HSC in pooled samples, UCB pools were prepared containing 2 to 4 UCB samples. CD34+ HSC were isolated by MACS and analyzed by flow cytometry and antibodies to human CD45, CD34 and CD3, with and without ISHAGE gates. Samples of CD34+ HSC purified from a single UCB collection were analyzed concurrently to give the background value for co-purification of syngeneic T cells and formation of T:HSC aggregates in the CD34-selected products. We found a limited number of CD3+ T cells present in CD34+ HSC isolated from single UCB collections [mean = 0.25%, range = 0 – 1.1%]. That value was slightly elevated when pools of UCB were used [mean = 0.44%; range = 0 – 1.9%]. There are few, if any, CD34+/CD3+ cells that can be detected by either the standard ISHAGE gating or by gating merely on CD45+ cells with no side scatter gating [0.02 – 0.04%]. This was true of CD34+ HSC isolated from either pooled UCB or single UCB collections.
Based on these results, we conclude that there is not significant co-purification of CD3+ T cells with CD34+ UCB-HSC, and that any such complexes that form are not found at any greater frequency in UCB pools than in single UCB collections.
Optimum Temperature for Maintaining the Viability of CD34+ Cells during Storage and Transport of Fresh Hemopoietic Progenitor Cells.
Authors: Antonenas Vicki , Garvin Frances , Webb Melina , Sartor Mary , Bradstock Kenneth F. , Gottlieb David J. 
Published: April 2020

Abstract
Background and Aim: The optimum storage and transport of freshly harvested hemopoetic progenitor cells (HPC) in the liquid state is not specified in the JACIE and FACT guidelines. Depending upon transplant centre, there is a range of reported ideal temperatures (1°C to 24°C) for HPC storage and transport but little data exists to justify the recommendations. Due to the limitations of Trypan Blue viability assays and CFU-GM colony assays, we used a no-lyse, CD34 assay (Sartor et al, Bone Marrow Transplantation 2005) to determine the optimum storage and transport temperature for maintaining viability of CD34+ stem cells in freshly harvested HPC.
Method: Samples were aseptically removed from 46 fresh HPC harvests (34 PBSC & 12 BM) and stored at refrigerated temperature (2°– 8°C), room temperature (18°– 24°C) and 37°C, for up to 72 hours. Samples were analysed for viable CD34+ cells/ml at 0, 24, 48 and 72 hours.
Results: The mean viable CD34+ yield prior to storage was 7.7x106/kg (range: 0.7 – 30.3). No viable CD34+ cells remained after storage at 37°C for 24 hours. The mean % loss of viable CD34+ cells at refrigerated and room temperatures can be summarized as follows:
Conclusion: These results demonstrate that the optimum temperature to maintain the viability of CD34+ stem cells for up to 72 hours during storage and transport of freshly harvested HPC is 2°– 8°C.
Mean % loss or gain of CD34+ cells on storage Time Refrigerated Temperature Room Temperature N Mean Range N Mean Range 24 hrs 39 −9.4 +15 to −42 23 −21.9 +17 to −59 48 hrs 31 −19.4 +7 to −69 20 −30.7 +3 to −68 72 hrs 29 −28.0 +4 to −53 18 −43.3 +11 to −70
Comparison of Umbilical Cord Collection Methods: Impact on Cellularity and Microbial Contamination Rates.
Authors: Foster Karen A. , Chasan-Taber Scott , Dracker Robert A. , Gunter Kurt C. 
Published: April 2020

Abstract
Introduction
Umbilical cord blood collections are typically performed using one of two methods: the syringe collection method and the gravity collection bag method. Collection volume, total nucleated cell (TNC) count, total CD34 positive cell count and percent with microbial contamination were analyzed to determine differences between collections performed using these two methods.
Methods
Units were processed using a hydroxyethyl starch red blood cell depletion method. TNC counts were measured using a Beckman Coulter HmX Analyzer. Total CD34 positive cell counts and viability were measured using the Beckman Coulter Epics XL-MCL flow cytometer. Microbial testing was performed using the VersaTrek system. Data related to collection volume and cellular content were analyzed by a Wilcoxin rank sum test of the paired differences [95% confidence interval (CI) for the difference between the control and experimental (syringe collections) results].
Results and Conclusions
The total cord blood volume collected via the syringe method was not significantly lower than that collected using the gravity collection bag method. However, the TNC count and total CD34 positive cell count from syringe collections were significantly inferior to the TNC count and CD34 positive cell count from gravity bag collections. The rate of microbial contamination of units collected in syringes was 3.3-fold higher than that of units collected using gravity collection bags. Cord blood banks should train collection sites on optimal collection methods and monitor the compliance of all collection sites with standardized collection procedures.
Summary of Results Gravity Bag Collections Syringe Collections Collection Volume N 27,121 55 Mean 66 mL 60 mL Standard Deviation +/− 30 mL +/− 24 mL Wilcoxin rank sum test (p-value) 0.0945 TNC Count (Post Processing) N 27,121 55 Mean 7.5 x 10^8 5.0 x 10^8 Standard Deviation +/− 4.6 x 10^8 +/− 3.5 x 10^8 Wilcoxin rank sum test (p-value) &lt;0.0001 Total CD 34 Positive Cells (Post Processing) N 20,495 51 Mean 2.76 x 10^6 1.77 x 10^6 Standard Deviation +/− 2.87 x 10^6 +/− 1.32 x 10^6 Wilcoxin rank sum test (p-value) &lt; 0.0001 Microbial Contamination Rate 7.1 % 23.6%
Pegfilgrastrim Compared with Filgrastim after Autologous Hematopoietic Peripheral Blood Stem Cell Transplantation.
Authors: Vanstraelen Gaetan , Frère Pascale , Ngirabacu Marie-Christine , Willems Evelyne , Fillet Georges , Beguin Yves 
Published: April 2020

Abstract
In order to assess the effect of Pegfilgrastim on the duration of neutropenia and clinical outcome of patients after autologous peripheral blood stem cell transplantation (PBSCT), we compared 20 consecutive patients with lymphoma or multiple myeloma receiving a single 6 mg dose of Pegfilgrastim on day 1 posttransplant to a historical control group of 60 patients receiving daily Filgrastim 5 μg/kg starting on day 1 posttransplant. There were 54 M and 26 F, 30 patients with lymphoma and 50 with myeloma, 26 in CR and 54 not in CR. Mean age was 55±10 yrs and 25 had already received a previous autologous transplant. The two groups were matched for disease and disease status, transplant number, age and sex. Cell dose infused tended to be higher in the Pegfilgrastim group (7.16±3.82 vs 10.03±6.25 x106 CD34+ cells/kg, p=0.0575). There were no differences (p&gt;0.05) in time to 0.5 (8 vs 9 days) or 1 (9 vs 9 days) x109/L neutrophils; to 1 % reticulocytes (13 vs 15 days) or 9 (12 vs 14 days) or 10 (30 vs 25 days) g/dL Hb; to 20 (9 vs 9 days) or 100 (20 vs 31 days) x 109/L platelets. The number of days with fever (2.7±2.3 vs 2.3±2.4 days), incidence of infections (all infections; bacteremia; bacterial, fungal or viral infections; FUO), duration of antibiotic therapy (8.7±5.9 vs 8.4±5.9 days), RBC (1.1±1.6 vs 0.9±1.6) and platelet (1.0±1.7 vs 1.2±1.8) transfusions, and time to hospital discharge (14.5±5.3 vs 15.4±5.8 days) were similar in the Pegfilgrastim compared to the Filgrastim group. However, after initial hematopoietic recovery, several differences between the groups became apparent, with the group always showing higher counts compared to the Filgrastim group (p values &lt;0.05 to &lt;0.001). Neutrophils remained significantly higher in the Pegfilgrastim group between days 14–30, lymphocytes between days 56–90, monocytes between days 21–24, reticulocytes between days 17–42 and platelets between days 35–90, respectively. These differences had no impact on clinical outcome after day 30 due to the low incidence of infectious events after engraftment. We conclude that Pegfilgrastim administrated on day 1 posttransplant facilitates early hematopoietic reconstitution comparable to daily Filgrastim. However, despite a trend towards fewer CD34+ cells transplanted, the Pegfilgrastim group enjoyed higher trilineage cell counts for some time after initial engraftment. This should be further tested in prospective randomized trials.
Successful Autologous Stem Cell Transplantation in Patients with Non-Hodgkin’s Lymphoma over the Age of 70 Years.
Authors: Yusuf Rushdia Z. , McAfee Steve , Dey Bimalangshu R. , Yeap Beow , Spitzer Thomas R. , Ballen Karen K. 
Published: April 2020

Abstract
Autologous stem cell transplantation (SCT) is a potentially curative procedure for patients with relapsed lymphoma (NHL). Over 60% of the cases arise in adults over 60 years of age. We retrospectively analyzed the outcomes of patients &gt;60 years old who received high dose busulfan (Bu) and cyclophosphamide (Cy) and autologous SCT for NHL. Stem cells were mobilized with G-CSF with or without chemotherapy. Patients received a dose-adjusted regimen of Bu 0.8 mg/kg IV (n=17) or 1mg/kg po (n=9) q 6 hours x 14 doses starting on Day -7 and Cy 60 mg/kg IV q day on days -3 and -2. In overweight patients, IV Bu doses were calculated based on ideal body weight and oral Bu doses and Cy doses were calculated on an adjusted body weight. All patients were treated as inpatients and received our usual supportive care including transfusion support, pneumocystis prophylaxis, antiviral prophylaxis, growth factor support and broad-spectrum antibiotic coverage for febrile neutropenia. 26 consecutive patients &gt; 60 with relapsed NHL underwent autologous SCT at Massachusetts General Hospital between 1995 and February 2005. The median age of the population was 66 (range 60–78) years. 17 (65%) of the 26 patients were females. 15 patients had large cell lymphoma, 2 immunoblastic lymphoma, 3 follicular lymphoma, and 6 mixed small cleaved cell and large cell. All patients had recurrent disease; 7 patients were in a CR and 19 patients in a PR at the time of transplant. The median time from diagnosis to transplant was 32 months. All patients engrafted. The median time to absolute neutrophil count &gt; 500/uL was 10 days (range 9–11) days. The median time to platelet count &gt; 50,000/uL unsupported was 15 (range 12–39) days. The 100-day transplant related mortality was 0%. Patients experienced the following serious toxicities: interstitial lung disease (n=1), seizures in a patient with CNS lymphoma (n=1), venoocclusive disease which resolved (n=2), and transient atrial fibrillation (n=3). With a median follow-up of 22 months (range 4 months–9.5 years), the two year overall survival is 63% and two year progression-free survival is 42%. Six patients above the age of 70 years have been transplanted. 100-day transplant related mortality was 0%. With a median follow up of 25 months, all 6 patients are alive and 3 of 6 are progression free. In conclusion, 1) Bu/Cy is a well-tolerated conditioning regimen for patients with NHL above the age of 60 years, including selected patients over the age of 70; 2) Age alone should therefore not be used as an exclusion criterion for autologous SCT.
Subclinical Alterations in Coagulation in Patients during Conditioning Regimen before Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Jiang Qian , Huang Xiao Jun , Liu Kaiyan , Chen Huan , Chen Yuhong , Cao Zhiyong , Lu Daopei 
Published: April 2020

Abstract
Objective To evaluate the alterations in coagulation in patients during modified busulfan plus cyclophosphamide (BUCY) ± antithymocyte globulin (ATG) before allogeneic hematopoietic stem cell transplantation (allo-HSCT), and to assess the effect of ATG on coagulation system as part of conditioning regimen.
Methods Thirty-five patients with various hematological malignancies undergoing allo-HSCT were assessed. Nineteen patients from HLA-identical siblings (group A) were conditioned with modified BUCY regimen, included cytarabine (2g/m2 i.v., day -9), busulfan (4mg/kg p.o. in divided doses daily, day -8 to day -6), cyclophosphamide (1.8g/m2 i.v., day -5 and day -4) and Me-CCNU (250mg/ m2 p.o., day -3). Sixteen patients from HLA-mismatched family members or HLA-matched unreleated donors (group B) were conditioned with modified BUCY + ATG regimen, added cytarabine (4g/m2 i.v., day -10 and -9) and rabbit ATG (2.5mg/kg i.v., day -5 to day -2, SangStat S.A.S., France). Blood samples were obtained before the start of regimen until day +1 after allo-HSCT. The following laboratory parameters were measured: prothrombin time (PT), active partial thromboplastin time (APTT), Fgrinogen (Fg), antithrombin (AT), D-Dimer, Fgrin degradation product (FDP), platelet (PLT), liver enzymes and bilirubin. VIII:C, IX:C, XI:C and XII:C in some blood samples with prolonged APTT were determined. Clinical hemorrhagic symptoms were monitored.
Results From day -5 of conditioning regimens, temporary lengthening of APTT, which peaked on day -3, occurred in 16/19 (84.2%) patients in group A and 19/19 (100%) patients in group B, continued rise in Fg occurred in 17/19 (89.5%) patients in group A and 19/19 (100%) patients in group B, a progressive decrease of PLT was observed in all patients of two groups. Alterations of Fg and PLT were more significant in group B compared to those in group A. Transient D-Dimer increase was detected only in group B on day -3. Among intrinsic pathway coagulation factors, XII:C and XI:C were decreased commonly and significantly when APTT was prolonged. No difference between the two groups could be found with regard to PT, FDP, AT and liver parameters which remained nearly in normal ranges. Most of patients in two groups did not have overt bleeding manifestations.
Conclusions Modified BUCY ± ATG conditioning regimen can induce subclinical alterations in coagulation. The regimen contained ATG has more significant effect on coagulation parameters.
Pharmacokinetics (PK) of 2 Dosing Regimens of Palifermin in Patients (Pts) with Hematologic Malignancies (HM) Undergoing High-Dose Chemoradiotherapy and Hematopoietic Stem Cell Transplantation (HSCT).
Authors: Zia-Amirhosseini Parnian , Salfi Margaret , Elhardt Dieter , Aycock Jeff , Cheah Tsui Chern , Cesano Alessandra , Hurd David D. 
Published: April 2020

Abstract
For pts with HM undergoing HSCT, oral mucositis (OM) is a frequent and debilitating complication that negatively impacts treatment outcomes, patient quality of life, and healthcare resources. Palifermin reduces the incidence and duration of severe OM in the HSCT setting. This phase 1 open-label study assessed the PK of 2 palifermin dosing regimens. Methods: Pts were 18 to 76 years old with HM and a Karnofsky performance score ≥70%. Palifermin was administered intravenously once daily as follows: 60 mcg/kg/day [d] for 3 consecutive days on d -11, -10, and -9 before conditioning (total body irradiation [TBI] + etoposide + cyclophosphamide) and following HSCT on d 0, 1 and 2 (part A) and a single dose of 180 mcg/kg (part B) before conditioning on d -11 and after HSCT on d 0. In part A (6 total doses), PK parameters were assessed after the 1st, 3rd, 4th, and 6th doses. In part B (2 total doses), assessments were made after each dose administration (d -11 and d 0). Results: In part A, 13 pts received palifermin; in part B, 12 pts received the single dose on d -11 and 11 pts received the single dose on d 0. For both dosing regimens, palifermin concentrations declined rapidly (≥98% decrease) in the first 30 minutes postdose, followed by a slight increase in mean concentrations between 1 and 4 hours and then a terminal decay phase. Respective mean (SD) PK parameter values for the 2 dosing regimens are shown in Table 1. In part A, mean AUC0-t values were comparable between doses 1 and 3 (within 15%) and 1 and 4 (within 1%). In part B, mean PK parameter values were similar (within 10% of each other) between doses 1 and 2. The mean AUC after the first 180 mcg/kg dose in part B was approximately 4-fold higher than that after the first 60 mcg/kg dose in part A. Mean half-life values ranged between 3.3 to 5.7 hours in part A and the value was 5.4 hours in part B. Conclusion: The PK data in pts receiving HSCT were consistent with approximately dose-linear PK in the dose range of 60 and 180 mcg/kg, with no observed accumulation, based on AUC, after 3 daily doses of 60 mcg/kg in this pt population in the HSCT setting.
Table 1 Dose Number (Dosing Day) n AUC0-t (hr x ng/mL) mean (SD) Clearance (mL/hr/kg) mean (SD) Vss (mL/kg) mean (SD) a Accurate computations of clearance (CL) and volume of distribution at steady state (Vss) were not possible for some concentration-time profiles. Part A - 60 mcg/kg/day x 3 consecutive days 1st dose (day -11) 9–13 34.3 (15.9) 1730a (497) 5320a (2330) 3rd dose(day -9) 13 39.8 (36.4) - - 4th dose(day 0) 11–13 34.8 (22.5) 2030a (862) 3870a (2080) 6th dose(day 2) 13 21.2 (15.1) - - Part B - 180 mcg/kg/day x 1 day 1st dose(day -11) 12 140 (50.9) 1460 (600) 4290 (3270) 2nd dose(day 0) 11 143 (71.8) 1770 (1290) 4270 (4700)
Developing an Algorithm for Monitoring Chimerism after Allogeneic Bone Marrow Transplantation - A Single Centre Experience.
Authors: Chandy Mammen , Shanmugapriya S. , Balasubramaniam Poonkuzhali , SatheeshKumar S. , Shaji R.V. , Mathews Vikram , George Biju , Vishwabandya Auro , Srivastava Alok 
Published: April 2020

Abstract
Analysis of chimerism for the evaluation of engraftment and rejection after allogeneic stem cell transplantation is now standard practice (ASCT). The most common technique used is PCR amplification of short tandem repeats (STRs) and variable number of tandem repeats (VNTRs). Data on informativeness of various VNTRs or STRs in HLA identical recipient donor pairs in a population is essential to establish a scheme of sequential testing. To determine the informativeness in our population, we screened five STRs namely FES, THO 1, VWA, ACTBP2, and F13A1 and 1 VNTR, APOB, in allogeneic bone marrow transplant patients with their HLA identical donors. Samples from 310 patients who underwent haematopoietic stem cell transplantation (HSCT) during the period 1993–2005 were analyzed. All donors were HLA identical siblings or related family members of the patient. Of the 310 patients, 120 (39%) were from southern India, 88 (28%), 25(8%), and 59 (19%) from northern, eastern and western parts of India respectively. Seventeen (5%) patients were from other countries (Maldives (n=4), Oman (n=4), Sri Lanka (n=2), Singapore (n=1), Mauritius (n=1), Pakistan (n=3), Bhutan (n=2)). For 15 (5%) of these 310 patients only amelogenin PCR results were available. The other 295(95%) patients and donors were studied by PCR for one or more of the polymorphic repeat sequences, (STRs or VNTR) namely FES, THO 1, VWA, ACTBP2, F13A1 and APOB. Of all the six markers used THO1 was the most informative (55%) and F13A1 the least (20%). Informativeness of the markers THO1+VWF, THO1+VWF+ACTBP2, THO1+VWF+ACTBP2+FES was 79, 87, and 92 percent respectively. By combining one, two or three markers, we could discriminate between 68 to 92 percent of the patient donor pairs. Addition of marker APOB to this panel discriminated 95% of the HLA identical patient donor pairs. Usage of Genescan analysis in practice to evaluate chimerism status has improved the utility of these markers from 95% to 97%. Using a combination of all these markers along with a polymorphic marker in the b-globin gene and Y chromosome specific amelogenin marker, we were able to discriminate 99% of the patient donor pairs. We have established an algorithm for evaluating chimerism in the Indian population using molecular markers in 310 patients and their HLA identical related donors. This data may be useful for transplant centers in adopting a strategy for evaluating chimerism following HLA identical ASCT.