bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clonal Involvement of the CD34+CD33− Progenitor Population by Leukemic Cells Harboring FLT3/ITD Correlates with High Risk of Relapse in Pediatric Acute Myeloid Leukemia.
Authors: Wright Jessica A. , Alonzo Todd A. , Gerbing Robert B. , Woods William G. , Lange Beverly J. , Radich Jerald P. , Bernstein Irwin D. , Meshinchi Soheil 
Published: October 2019

Abstract
Internal tandem duplication of the FLT3 gene (FLT3/ITD) has been associated with high risk of relapse in acute myeloid leukemia (AML) yet nearly 25–30% of the patients with FLT3/ITD have long-term disease free survival with conventional chemotherapy. We hypothesized that FLT3/ITD AML patients with poor clinical outcome may have disease that involves less mature hematopoietic precursors than patients with favorable outcome. To test this hypothesis, we isolated less mature, CD34+CD33− and more mature, CD34+CD33+ precursor cells from 24 pediatric AML patients enrolled on Children’s Cancer Group clinical trials CCG-2891 and 2961 previously identified as having a FLT3/ITD. Granulocyte/monocyte colonies (CFU-GM) were grown in methylcellulose, harvested, and analyzed for the presence of FLT3/ITD after 14 days of growth. Twenty patients yielded sufficient cells and growth of CFU-GM colonies for analysis. FLT3/ITD was detected in CFU-GM colonies derived from CD34+CD33+ cells in all patient samples (median 80% of colonies tested per patient, range 6–100%). In contrast, FLT3/ITD was detected in CFU-GM colonies derived from CD34+/CD33− cells in only 11 of the 20 patient samples (median 46% of colonies tested per patient, range 6–100%). Of the 9 patient samples without FLT3/ITD involvement of CD34+CD33− colonies, 8 achieved a CR, 6 of whom are long-term survivors, and one patient died of non-leukemic causes. In contrast, of the 11 patients with CD34+CD33− cell involvement, 9 either failed to achieve CR or relapsed after achieving CR, and 2 died of non-leukemic causes. Actuarial progression-free survival at 4 years from diagnosis for the patients with and without FLT3/ITD in the CD34+CD33− population was 0% vs. 68% respectively (p=0.017). As allelic ratio of the FLT3/ITD has been used to define high-risk patients within the FLT3/ITD cohort, we determined the FLT3/ITD allelic ratio in our study population and correlated it with the presence of FLT3/ITD in the CD34+CD33− population. Ten of the 11 (91%) of the patient samples with FLT3/ITD involvement of the progenitor cells had high allelic ratio compared to 5 of 9 (56%) of the patients without early cell involvement. Together these data suggest that clonal dominance of FLT3/ITD containing leukemia cells at the CD34+CD33− stage of hematopoietic development is correlated with a high risk of relapse. Further studies are required to determine whether clonal dominance at this hematopoietic stage is a variable that, independent of high allelic ratio, accounts for the poor clinical outcome seen in a subset of FLT3/ITD positive AML patients.
Homeostatic Proliferation of Chronic Lymphocytic Leukemia.
Authors: Wirths Stefan , Lanzavecchia Antonio 
Published: October 2019

Abstract
Slow accumulation of chronic lymphocytic leukemia cells in vivo was considered due to defective apoptosis rather than proliferation. However, recent data, based on isotope incorporation studies, suggested significant continuous proliferation of CLL cells in vivo and similar observations has been made for human memory B cells. As previous gene expression studies revealed close relation of CLL cells to memory B cells we asked, whether the mechanisms driving homeostatic proliferation of human memory B cells were stimulating CLL cells as well. Comparison of CLL samples (n=20) and memory B cells of healthy donors showed high expression levels of Toll-like receptor (TLR) 7 and TLR9 and of IL2 and IL15 cytokine receptors compared to naive B cells. Proliferation of CLL cells compared to their normal counterparts was analyzed after FACS sorting and CFSE-labeling.Naive B cells did neither respond to TLR7 ligand resiquimod, TLR9 ligand CpG2006, IL2 and IL15 nor to their combination. In contrast, CLL cells and memory B cells showed comparable patterns of proliferation. Remarkably, under these conditions, terminal differentiation to plasma cells was observed for memory B cells only, while proliferation of CLL ceased after the 4th division without differentiation into Ig-secreting cells. To estimate in vivo turnover rates, B cells of healthy donors and CLL cells were stained for intracellular Ki67, that identifies recently divided cells, and were analyzed by flow cytometry. Memory B cells showed high in vivo turnover rates (2–5% Ki67+) while naive B cells where largely quiescent - results that are in line with previous isotope incorporation studies with healthy donors. Turnover rates of CLL ex vivo ranged from 0.025–1.4% Ki67+ cells, which in absolute numbers (per mL of blood) was comparable to memory B cells. These results are consistent with a concept of homeostatic proliferation of both memory B cells and CLL cells, the latter being arrested without terminal differentiation, which might account for their accumulation in vivo.
NK Cells Efficiently Prevent Engraftment of Donor Stem Cells after Tolerigenic Bone Marrow Transplantation.
Authors: Kean Leslie , Hamby Kelly , Pearson Thomas , Larsen Christian 
Published: October 2019

Abstract
Introduction: Immunologic tolerance remains an elusive goal of transplantation. In mice, mixed-chimerism and donor-specific tolerance can be induced by blocking the CD28/CD40L T-cell costimulatory pathways after bone marrow transplant (BMT). However, large doses of marrow (~1x109 cells/kg) are required, and these regimens have not yet been successfully translated to clinical practice. There is a growing body of evidence that NK cells may play a central role in the failure of low doses of donor bone marrow to engraft, but the mechanisms underlying NK alloreactivity remain to be determined.
Methods: (1) BMT in the presence of CD28/CD40L T cell costimulation blockade was performed using C57BL/6 (B6) recipients and Balb/C donor bone marrow. The role of host-anti-donor NK alloreactivity in preventing engraftment was determined by specifically depleting B6 NK cells. The contribution of the NK cell-surface receptor, LFA1 to NK alloreactivity was determined with the anti-LFA1 blocking antibody M17/5.2. (2) An in vivo NK alloreactivity assay was developed that should allow the investigation of the mechanism of NK alloreactivity and the molecular mediators of this process. In this assay, CFSE-labeled B6 splenocytes were adoptively transferred into B6xBalbC F1 progeny. As such, alloreactivity was specifically mediated by NK cells. NK alloreactivity was measured flow-cytometrically by the disappearance of the CFSE-labeled B6 population.
Results: Transient depletion of recipient NK cells resulted in increased donor stem cell survival and the induction of stable mixed-chimerism and tolerance despite BMT with low doses (≤2x106 cells) of donor bone marrow. This effect was specific to allogeneic donor cells: depletion of NK cells did not increase engraftment of syngeneic bone marrow. Blocking the adhesion molecule, LFA-1 recapitulated the effects of whole-scale NK depletion. Newly emergent NK cells exhibited significantly lower expression of the donor-specific activating receptor, Ly49D, and these NK cells did not exhibit in vivo alloreactivity. These results suggest that the NK repertoire in the mixed-chimeric setting exhibited donor-specific tolerance.
Using the in vivo hybrid resistance NK alloreactivity assay, we measured 80% NK-specific target killing 8 days after adoptive transfer. Significantly less killing occurred at 2, 4, and 6 days. Pre-sensitizing the recipient for 4 days increased the efficiency of killing—from 50% to 80%, suggesting a potent activation phenomenon required for efficient NK allorecognition and/or cytotoxicity.
Implications: These results reveal the importance of NK alloreactivity in the acquisition of mixed-chimerism after BMT at limiting stem cell doses, and suggest that clinical approaches to tolerance-induction transplantation may require mechanisms to control NK alloreactivity.
Serum Soluble Syndecan-1 Levels at Diagnosis Constitute an Independent Prognostic Factor of Survival in Myeloma That May Further Differentiate Patients within the ISS Stages.
Authors: Kyrtsonis Marie-Christine , Vassilakopoulos Theodoros P. , Papadogiannis Argyro , Tzenou Tatiana , Kalpadakis Christina , Antoniadis Antoine , Dimopoulou Maria N. , Siakantaris Marina P. , Kokoris Stella I. , Dimitriadou Evangelia M. , Angelopoulou Maria K. , Plata Ellen , Tsaftaridis Panagiotis , Pangalis Gerassimos A. 
Published: October 2019

Abstract
Syndecan-1 (CD138) is a heparin sulphate proteoglycan overexpressed on the surface of both normal and malignant plasma cells. Its role in multiple myeloma (MM) pathogenesis remains unclear. It may be shed from plasma cell surface and released in its soluble form (soluble syndecan-1: s-synd-1). Serum s-synd-1 levels have been shown to be a powerful prognostic factor of survival in myeloma patients. The role of s-synd-1 in the current prognostic classification systems requires further investigation. We have retrospectively determined soluble syndecan-1 in frozen sera of 99 MM patients, 18 MGUS patients, and 25 healthy individuals (HI). In all patients sera samples were collected at diagnosis. Serum s-synd-1 levels were determined by ELISA (Diaclone Research, Besanson, France) according to the manufacturer’s instructions. MM patients median age was 69 y (44–82), 51 were female, 48 male, 26 were Durie-Salmon (DS) stage I, 41 stage II, and 32 stage III while, according to the International Scoring System for MM (ISS), 32 were in stage 1, 25 in stage 2 and 42 in stage 3. Median serum s-synd-1 was 15 ng/ml (7–40) in HI, 16.5 ng/ml (8–220) in MGUS and 64 ng/ml (7–3500) in MM. Differences between HI and MM patients levels and between MGUS and MM patients levels were both significant (0.01 and 0.05 respectively). Overall survival was decreased in patients with elevated (mean + 2SD) serum s-synd-1 (5-year overall survival 35±9 vs 80±13, p=0.004). S-synd-1 levels were an independent prognostic factor of survival compared to DS and ISS staging systems. In multivariate analysis, only ISS and s-synd-1 were significant parameters of prognosis (the prognostic significance of DS, ISS and s-synd-1 was 0.06, 0.006 and 0.02 respectively). Further analysis of survival of patients stratified according to the ISS system showed a difference in survival within groups according to s-synd-1 levels. The 5-year overall survival was 100 vs 58±16 in ISS stage 1 patients with low or elevated s-synd-1 respectively, 100 vs 18±16 in stage 2 and 50±25 vs 14±13 in stage 3. The difference was statistically significant for stage 2 only (p=0.04) and there was only a trend due to the low number of patients in stages 1 and 3. In conclusion serum s-synd-1 levels at diagnosis constitute an independent prognostic factor of survival, able to further differentiate MM patients within the ISS stages.
A Phase I Study To Investigate the Pharmacokinetics (PK) and Pharmacodynamics (PD) of an Oral Platelet Growth Factor (SB-559448) in Healthy Subjects.
Authors: Su Sheng-Fang , Peng Bin , Williams Daphne , Park Jung Wook , Handel Malcolm L. , Jenkins Julian , Ho Peter 
Published: October 2019

Abstract
Introduction: SB-559448, is an orally bioavailable, non peptide, small molecule thrombopoietin receptor agonist that induces proliferation and differentiation of megakaryocytes and progenitor cells, ultimately increasing the production of platelets.
Method: In a randomized, single blind, placebo-controlled, parallel group, phase I study in healthy subjects, SB-559448 was administered as oral capsules once daily for 1 day and, after a 7-day washout, for 10 days at doses of 160 to 640 mg. In this dose escalation study, subjects were randomized into 4 groups of 9 subjects to receive either active or placebo medication in a ratio of 6:3. Serial blood samples were collected to determine SB-559448 concentrations and estimate PK parameters following single- and repeat-dosing. PD assessments were performed every 4 to 5 days up to 31 days following initiation of repeat dosing.
Results: The preliminary results showed that SB-559448 was well tolerated in the study, there were no serious adverse events, no significant changes in laboratory or cardiovascular safety parameters and there was no observed relationship between the incidence or severity of adverse events and dose. Most adverse events were mild in intensity and self-limiting. SB-559448 was also shown to be orally bioavailable in humans with a pharmacokinetic profile suitable for a once daily oral dosing. When administered at oral doses of 160 mg and above for 10 days, a dose dependent increase in the platelet count was observed with maximum platelet count observed on days 14 to 15 following initiation of repeat dosing.
Conclusion: The results suggest that SB-559448 is an orally administered agent that increased platelet counts in healthy subjects. Additional studies will be needed to evaluate its effects in individuals with thrombocytopenia.
Inter-Laboratory Variation and the Effect of the Type of Fluorimeter on the Results of the Technothrombin® Thrombin Generation Assay.
Authors: Meijer Piet , Kluft Cornelis 
Published: October 2019

Abstract
Thrombin generation is an essential step in the clotting system. Therefore the measurement of thrombin generation is a desirable parameter to elucidate deviations in the clotting system. However till recently the measurement of thrombin generation was a difficult and laborious laboratory method. Nowadays commercial tests are available at the market. We evaluated in a multicenter trial the Technothrombin® TGA assay from Technoclone GmbH Vienna, Austria with a focus on inter-laboratory variability. The Technothrombin® TGA assay is a fluorogenic method. To our knowledge this was the first large-scale inter-laboratory comparison of a thrombin generation test. A set of 8 different samples were distributed to 45 clinical centers in Europe. Thirty-six returned results (80%). Samples were selected on a different level of thrombin generation as indicated by the thrombin concentration at peak height (PTC). The PTC in the samples used varied from 4 nM to 335 nM. The inter-laboratory variability varied from 16% for high PTC (~335 nM) to about 100% for low PTC (~4 - 15 nM).
Using a normal citrated pooled plasma as a reference the other samples can be significantly discriminated by all participants (P < 0.03) using an activator with a low phospholipid concentration (0.32 μM) in the test. With a high phospholipid concentration (3.2 μM) two of the samples can not be significantly discriminated from normal pooled plasma.
Furthermore, we evaluated the effect of the type of fluorimeter used by the clinical laboratories. The most frequently used fluorimeters in this study were the Biotek FLx 800 (n=11), Thermo Fluoroskan (n=8), Tecan Genios (n=5) and the BMG Fluostar (n=4). Overall there are no significant differences between the measurement of the PTC by the different fluorimeters. However remarkable differences were observed for the inter-laboratory variability between the different fluorimeters. The average coefficient of variation of all samples increased from BMG (19.3% ; range 11.1 - 26.9%) < Biotek (28.8% ; range 15.9 - 59.3%) < Tecan (38.5% ; range 12.6 - 81.5%) < Thermo (49.7% ; 19.1 - 96.6%).
This data shows the impact the selection of a type of fluorimeter may have on the comparability of test results of clinical samples. Selection of a measurement system with the lowest inter-laboratory variation in clinical studies shows a benefit for the comparability of study results.
Efficacy and Safety of Tinzaparin Sodium as a Bridging Agent for Patients on Oral Anticoagulant Undergoing General Surgery: Clinical Experience with the First 100 Consecutive Patients.
Authors: Volteas Spiridon K. , Babos Anastasios 
Published: October 2019

Abstract
The aim of the present prospective study was to test the efficacy and safety of a bridging protocol in patients under oral anticoagulants (OA) who need to undergo general surgery. One hundred consecutive patients receiving oral anticoagulant accenocoumarol for various cardiovascular disorders entered the study. Patients underwent 107 scheduled operations (among them 41 cholecystectomies, 24 colectomies, 13 hernia repairs and 9 ERCPs, the 7 followed by cholecystectomy), receiving the low molecular weight heparin (LMWH) tinzaparin sodium as the bridging agent. At least 4 days prior to the operation the OA was replaced with tinzaparin at a daily dose of 175 anti-Xa IU per kg of body weight. Patients received no LMWH on the day of the operation and the OA was co-administered with tinzaparin on the second post-operative day and for at least 2 days. When target INR (2.5 to 3) was achieved, tinzaparin administration was stopped. Lower limbs color duplex was performed pre-operatively and on the 2nd, 8th–10th and 20th–30th post-operative day. Complete anticoagulation profile, intra-operative bleeding and post-operative blood loss, large haematoma development (over 5 cm in diameter), deep vein thrombosis (DVT) and 30-day morbidity and mortality were thoroughly recorded. During the 30-day period 1 patient died (from sepsis), 9 developed wound infections and three underwent re-operations (two for anastomotic leaks and one for duodenal perforation following ERCP). None of these events was related to the bridging process. There were no proximal DVTs. Three asymptomatic distal vein thromboses (2 posterior tibial and one peroneal ) were identified on post-operative duplex scan. No large wound haematomas were seen. One colorectal surgery patient had a drain blood loss of 460 ml on the first and 140 ml on the second post-operative day, requiring blood transfusion; tinzaparin administration was delayed for three days in this case. Finally, in 17 cases we had difficulties in adjusting INR levels to the desired range; in these cases tinzaparin administration was prolonged for 2–5 days. We conclude that our bridging protocol, using therapeutic doses of the LMWH tinzaparin sodium as the bridging agent, is both safe and effective for replacing OAs in patients undergoing general surgery operations.
Thalassemia in a Hawaiian Family of Filipino Extraction
Authors: LAZAR HAROLD P. 
Published: October 2019

Abstract
Hematologic data are presented on thalassemia in a Hawaiian family of Filipino extraction, and support is given the recent trend to broaden the ethnic concept of thalassemia. Because of recent reappraisals of older reported cases by other investigators in the light of newer electrophoretic technics, the need for continued re-evaluation is suggested.
The Effect of Repeated Doses of Urethane (Ethyl Carbamate) on the Mitotic Activity and Cellular Composition of the Bone Marrow of Normal Mice
Authors: ROSIN A. , GOLDHABER G. 
Published: October 2019

Abstract
A single dose of urethane produced a rise in the mitotic index, due essentially to a large increase in the metaphase index in the bone marrow of normal mice. Repeated daily injections (three and six doses) produced an elevation of the mitotic index, which was, however, smaller than that observed after a single dose of urethane. After nine doses of urethane the mitotic index was found approximately at the same level as in the controls. The mitotic index in the bone marrow of urethane-treated mice never decreased below the normal level found in untreated mice.
After three, six and nine doses of urethane a rise in the percentage of ana- and telophases was found, in contrast to the increase in the percentage of metaphases after administration of a single dose of urethane. The percentage of prophases was decreased after six and nine doses of urethane.
The morphological changes in the mitotic figures were the same as those described after a single dose of urethane. They consisted mainly in bridge formation and chromosome aberrations in anaphase.
Very large promyelocytes, often with prematurely lobated or bizarre shaped nuclei, were observed.
There was damage to the nuclei of the erythroid precursors and consequently an increase in the myeloid-erythroid ratio in the bone marrow after repeated doses of urethane, especially after nine doses.
The maturation rate of the segmented leukocytes in the bone marrow of the urethane treated mice was augmented after repeated doses.
Studies on Leukemia
Authors: SANDBERG A. A. , CARTWRIGHT G. E. , WINTROBE M. M. 
Published: October 2019

Abstract
1. The urinary excretion of uric acid was studied in 17 normal subjects and in 38 patients with leukemia. The mean excretion of uric acid by the normal subjects was 6.5 mg./Kg. of body weight /24 hrs. The mean excretion of uric acid in 14 patients with acute lymphoblastic leukemia was 30.3 mg./Kg. /24 hrs.; in 13 patients with acute myeloblastic leukemia, 13.0 mg.; in 6 patients with chronic lymphocytic leukemia, 5.2 mg.; and in 5 patients with chronic myelocytic leukemia, 13.5 mg.
2. Following therapy with cortisone, 6-mercaptopurine or Amethopterin, the urinary excretion of uric acid increased in the cases of acute leukemia as the leukocyte count declined. As the leukocyte count approached normal levels, the uric acid excretion decreased. The urinary excretion of xanthine and guanine paralleled the excretion of uric acid.
3. In association with the administration of an aromatic nitrogen mustard derivative to one patient with chronic lymphocytic leukemia and Myleran to one patient with chronic myelocytic leukemia, there was only a slight increase in uric acid excretion.
Time Pattern of Vitamin B12Co60 Urinary Excretion in Man After Oral Administration and Parenteral "Flushing"
Authors: BEST WILLIAM R. , LANDMANN WENDELL A. , LIMARZI LOUIS R. 
Published: October 2019

Abstract
Serial urine collections in a number of patients with pernicious anemia given 2 µg B12Co60 orally followed in two hours by 1000 µg nonradioactive vitamin B12 showed little urinary radioactivity at any time. When these tests were repeated together with a potent oral dose of intrinsic factor concentrate, there was little activity during the first four hours. Peak excretion rates occurred most commonly between 6 and 12 hours after ingestion of radioactive B12, sometimes even later. The time of peak excretion was fairly characteristic for the individual. Secondary peaks occasionally occurred, and only slight radioactivity usually remained after 24 hours. It is postulated that the delayed peak is related to the time it takes for B12 to be transported in the intestine to the point of absorption or to the duration of the intracellular metabolic processes of absorption. For most purposes the use of fractional urinary collections is not necessary.
A Modification of the Benzidine Method for Measurement of Hemoglobin in Plasma and Urine
Authors: CROSBY WILLIAM H. , FURTH FRANK W. , THIBEAULT CAROL E. 
Published: October 2019

Abstract
A modification of the benzidine method for measuring hemoglobin in plasma and urine is described. When a known amount of hemoglobin is added to plasma or urine more than 95 per cent can be recovered with this procedure.
Studies on the Fibrinogen, Dextran and Phytohemagglutinin Methods of Isolating Leukocytes
Authors: SKOOG WILLIAM A. , BECK WILLIAM S. 
Published: October 2019

Abstract
Studies are presented on the dextran, fibrinogen, and phytohemagglutinin methods of leukocyte separation. Optimal conditions, obtainable yields, detailed directions, advantages and disadvantages of each procedure are presented. It is shown that by using these methods in combinations, leukocyte suspensions of extreme purity may be obtained.