bluuebunny/crossref_metadata_2025_split huggingface dataset:
Early Fludarabine Treatment of TCL-1 Transgenic Mice Promotes Drug Resistance through a Mechanism Not Involving p53 Mutations: Implications for Patients with Chronic Lymphocytic Leukemia (CLL).
Authors: De Lay Michael D. , Johnson Amy J. , Lucas David M. , Durr Darius T. , Sass Ellen M. , Goettl Virginia M. , Lehman Amy , Jarjoura David , Grever Michael R. , Byrd John C. 
Published: October 2019

Abstract
Initial therapy for Chronic Lymphocytic Leukemia (CLL) typically includes nucleoside analogs such as fludarabine. Mutations of p53 are uncommon in CLL patients at diagnosis, but are found at increased frequency in patients with prior treatment and are often associated with resistance to standard therapies including fludarabine. The causality of fludarabine treatment to p53 mutation has not been clearly defined. In previous work with the TCL-1 model of CLL, we demonstrated that mice with active leukemia do not have p53 mutations and respond to fludarabine therapy with a modest survival advantage (Johnson et al, Blood108:1334, 2006). We therefore initiated a randomized trial of fludarabine treatment (34mg/kg daily intraperitoneal injection for five days every 28 days) versus observation (treatment with volume-matched vehicle controls on the same dosing schedule) in TCL-1 transgenic mice prior to the onset of leukemia. The goals of this study were to determine if early treatment with fludarabine would 1) delay disease onset; 2) eventually yield a fludarabine-resistant phenotype; or 3) promote development of p53 mutations in mice developing progressive leukemia. Transgenic TCL-1 mice between the ages of eight and 12 weeks were continually treated with fludarabine (n=32) or saline (n=45) until death while being screened monthly for disease progression through changes in WBC count by peripheral blood differentiation smear, lymphocyte subsets by flow cytometry, and nodal and spleen exams. Despite continued treatment with fludarabine, drug resistance as manifested by leukemia and eventual death occurred. Median time to leukemia onset was 308 days (95% CI: 224, ∞) in the fludarabine group and 338 days (95% CI: 252, ∞) in the saline group. Thus, the risk of developing leukemia was not significantly different between the two groups. Of interest, p53 mutations in exons 2–11 were not found in either group (n=10 each) at the time of death. Microarray studies are currently underway to discover other genes differentially expressed between treated and untreated mice that might contribute to fludarabine resistance and will be presented. In conclusion, our data demonstrate that early fludarabine treatment in the TCL-1 transgenic model of CLL does not prevent onset of leukemia, but instead results in disease resistant to fludarabine. The source of fludarabine resistance in this mouse model was not through acquisition of p53 mutations, but rather an alternative unidentified mechanism(s).
The Proliferative Response to CpG ODN Stimulation Predicts Disease Progression in B-Cell Chronic Lymphocytic Leukemia.
Authors: Laurenti Luca , Longo Pablo G. , Tarnani Michela , Gobessi Stefania , Sica Simona , Leone Giuseppe , Efremov Dimitar G. 
Published: October 2019

Abstract
Chronic lymphocytic leukemia (CLL) B-cells from certain patients exhibit marked proliferation following stimulation with unmethylated CpG-oligodeoxynucleotides (CpG-ODN). In a recent study we observed that CpG-ODN induced proliferation occurs primarily in leukemic B-cells from patients with progressive disease and unmutated immunoglobulin (Ig) VH genes. To further investigate whether the proliferative response following CpG-ODN stimulation can be used as a prognostic marker in CLL, we evaluated progression-free survival (PFS) and treatment-free survival (TFS) in a larger series of well characterized patients that had been followed for an extended period. In addition, we correlated the proliferative response with other prognostic factors, including Ig VH gene mutation status, ZAP-70 expression and the presence of “high risk” cytogenetic abnormalities. Analysis of 3H thymidine incorporation in purified CLL B-cells stimulated for 72 hours with CpG-ODN 2006 revealed a proliferative response in 23 (40%) of the 58 evaluated cases. Eighteen (78%) of the proliferating cases had unmutated and 5 (22%) had mutated (M) Ig VH genes, whereas among the 34 non-proliferating cases with available Ig VH gene data 8 (24%) had unmutated and 26 (76%) mutated Ig VH genes (P=<0.001 by chi-square test). ZAP-70 was expressed by 16 of the proliferating (70%) and 14 of the non-proliferating cases (40%), but this difference was not statistically significant (P = 0.053). The high-risk risk cytogenetic abnormalities del(17p13.1) and del(11q22.3) were detected in 6 of the 21 evaluated proliferating (29%) and in 3 of the 30 evaluated non-proliferating cases (10%) (P = 0.136). Median PFS was 25 months in the proliferating and 72 months in the non-proliferating subset (P=0.001). Similarly, median TFS was significantly shorter in the proliferating (30 months) than in the non-proliferating subset (114 months, P=0.001). In summary, these data confirm that the proliferative response to CpG-ODN stimulation is strongly correlated with Ig VH gene mutation status, but does not depend on ZAP-70 expression. Although high-risk cytogenetic abnormalities were more frequent in the proliferating subset, the difference was not significant. Cases with a positive proliferative response had significantly shorter PFS and TFS, indicating that the proliferative capacity of the leukemic cells is an important determinant of the clinical course in CLL. Elucidation of the mechanisms that underlie the different proliferative capacity of CLL B-cells should provide additional insight in the pathogenesis of CLL and may lead to the identification of novel and clinically useful prognostic markers.
Comparative Analysis of Coagulation System Proteins Distribution in Breast Cancer Primary Lesions and Lymph Node Metastases.
Authors: Wojtukiewicz Marek , Sierko Ewa , Sawicki Zbigniew , Zimnoch Lech , Kisiel Walter 
Published: October 2019

Abstract
Breast cancer is the most frequent cause of morbidity and mortality due to cancer in women. Thromboembolic episodes are relatively common complication during the course of the disease. It was widely documented, that components of blood coagulation play a role in tumor progression and metastasis formation. Presence of hemostatic proteins was observed in primary lesions of different malignant tumors. However, information on expression and role of coagulation system components in regional lymph node metastases is scanty. The purpose of the study was to assess and compare the distribution of selected coagulation proteins - in primary versus metastatic lesions of breast cancer. Specimens of primary tumor of different degree of malignancy and metastatic lesions localized in regional lymph nodes were obtained during surgical treatment of previously untreated twenty five breast cancer patients and fixed in formalin. Immunohistochemical procedure was employed using monoclonal and polyclonal antibodies against tissue factor (TF), tissue factor pathway inhibitor (TFPI), factor IX, factor X, fibrinogen and fibrin. A strong expression of TF was demonstrated in cancer cells of both primary tumors and lymph node metastases. Coagulation factors IX and X were also present in primary and metastatic breast cancer lesions. However, the expression of the proteins was weaker in regional lymph node metastases than in primary tumor. Strong staining for fibrinogen was detected in the stroma of primary breast cancer, particularly at the area adjacent to the cancer foci. Stroma of metastatic lesions was characterized by much weaker staining for fibrinogen. Strong expression of fibrin and TFPI was demonstrated in cancer cells of primary tumor while much weaker expression of the antigens was visualized in cancer cells localized in lymph node metastatic lesions. Furthermore, the presence of all examined proteins was observed in small blood vessel walls and in macrophages infiltrating both primary tumor tissue as well as metastatic lesions. The results of the study indicate extravascular activation of blood coagulation in loco in breast cancer, both in the primary tumor and in the regional lymph node metastatic lesions. The presence of coagulation system proteins in cancer tissue (primary tumor and metastatic foci) suggests that they may play a role in the biology of breast cancer growth as well as they may modulate breast cancer metastatic dissemination.
Effects of “Primary Prophylaxis” and “On-Demand” Therapy on the Clinical Expression of Severe Hemophilia A in Children - Results of a Multicenter Study.
Authors: Schobess Rosemarie , Kurnik Karin , Kreuz Wolfhart , Friedrichs Frauke , Krumpel Anne , Bidlingmaier Christoph , Ettingshausen Carmen Escuriola , Nowak-Gottl Ulrike 
Published: October 2019

Abstract
Background: Patients with severe hemophilia A (HA) can either be treated by regular FVIII infusions twice or thrice per week (prophylaxis), or only in case of bleeding episodes (on-demand). Whereas prophylaxis reduces the number of bleeding episodes and may therefore prevent the development of hemophilic arthropathy, recommendations regarding age and dose at start of prophylactic regimens are still a matter of debate. The present cohort study was performed to investigate the role of “primary prophylaxis” versus “on-demand” therapy in HA children. The outcome variable was imaging-proven hemophilic joint damage.
Methods: 42 children were initially treated with primary prophylaxis following the first bleeding episode, and were frequency-matched (year of birth, catchment area) to 67 patients receiving “on-demand” therapy with an early switch to “secondary prophylaxis”.
Results: In multivariate analysis adjusted for the HA mutation type and the presence or absence of thrombophilia, the Pettersson score investigated at a median age of 12.5 years in joints with at least one documented bleeding episode was no significantly different between the two patient groups (p=0.944), and no statistically significant differences were found in patients with target joints (p=0.3), or in children in which synovitis had occurred (p=0.77). Imaging results obtained showed a substantial agreement (87.14%) beyond that expected by chance alone (42.4%) between local and central readers in the patients tested (kappa=0.77; Z= 17.27; p < 0.001).
Conclusion: In cases with severe HA where primary prophylaxis is impossible, the procedure to switch from “on-demand” to early secondary prophylaxis can be achieved in the majority of young children affected. In addition, the needs of parents around the time of diagnosis of severe HA could be better addressed.
In Vitro and In Vivo Characterization of Full-Length rFVIII Modified with PEG Via Coupling to Primary Amino Groups.
Authors: Turecek Peter L. , Siekmann Jürgen , Gritsch Herbert , Váradi Katalin , Ahmad Rafi-Uddin , Muchitsch Eva-Maria , Ehrlich Hartmut , Schwarz Hans Peter 
Published: October 2019

Abstract
Chemical modification of recombinant therapeutic proteins with PEG has been shown to enhance the biological half-life. Here we assess the effect of PEGylation on FVIII. Full-length rFVIII bulk drug substance from protein-free fermentation (Advate process, Baxter) was conditioned into a buffer suitable for coupling to polyethylene glycol succinimidyl succinate (linear PEG, 5 kDa PEG chain length). PEG was covalently bound by amine coupling preferentially to lysine residues of FVIII at neutral pH. PEG was removed by ion-exchange chromatography and the PEG-FVIII derivative was concentrated by ultra-diafiltration. The conjugates thus obtained retained about 30–40% of the activity of non-modified rFVIII. The specific activity decreased with the amount of PEG linked to the FVIII molecule. In SDS-PAGE and immunoblot studies PEGylated rFVIII showed a band pattern similar to unmodified FVIII with full-length, heavy chain fragments of 180 kDa and 120 kDa and the light chain fragment of 80 kDa. PEGylation also occurred to a high extent in the B domain of FVIII. All bands appeared broadened due to the attachment of polymeric PEG. The maintenance of functionality of FVIII was demonstrated by its potential to be activated and inactivated by thrombin. In the assay PEGylated and unmodified FVIII were incubated with 1 nM thrombin. Sub-samples were drawn at intervals up to 40 minutes and added to a mixture of FIXa, FX, phospholipid vesicles and Ca2+ containing a thrombin inhibitor. After 3 minutes incubation at 37°C the amount of activated FX (FXa) was measured using a FXa-specific chromogenic substrate. Unmodified rFVIII showed a typical picture of an immediate increase in FXa activity and a subsequent decline with no further FXa generation after 15 minutes. PEGylated rFVIII was activated to the same extent as unmodified FVIII but the decay in FXa generation was slower and did not reach the zero level, even 40 minutes after incubation. The formation of the typical thrombin cleavage fragments, with unmodified as well as PEGylated rFVIII, was demonstrated in a Western blot analysis. The slower inactivation by thrombin was also seen there. The pharmacokinetic properties of PEGylated rFVIII compared with rFVIII were investigated in hemophilia A knock-out mice. Both preparations were applied at a dose of 200 IU rFVIII/kg and groups of mice (n=5) were exsanguinated at several time points up to 24 hours. Terminal half-life for PEGylated rFVIII was calculated at 4.9 hours compared with 1.9 hours for unmodified rFVIII in hemophilia A knock-out mice. AUC was approximately doubled. These results indicate that rFVIII can be biochemically modified with PEG whilst at least partly retaining its major functions, but at the same time prolonging its survival in the circulation of hemophilic mice.
Prevalence Estimates of Adult Chronic Idiopathic Thrombocytopenic Purpura (ITP) in the United States.
Authors: Feudjo-Tepie Maurilee A. , Robinson Noah J. , Bennett Dimitri 
Published: October 2019

Abstract
ITP is a disease caused by inadequate platelet production as well as increased platelet destruction. Literature on the epidemiology of ITP is currently sparse and the disease remains poorly described. Based on previous reviews, the overall incidence of ITP among adults ranges from 1.6 to 3.0 per 100,000 person years of observation (PYO); prevalence estimates range from 2.1 to approximately 36.4 per 100,000 persons (Neylon AJ et al., Br J Haematol. 2003; Satia J et al., Eur Hematol Assoc. [poster presentation] 2006; Deitz AC et al., Blood. 2006). In this study, the objective was to estimate the prevalence of adult chronic ITP (in 2005) stratified by age and gender using a large US claims database (Integrated Healthcare Information Services, IHCIS). IHCIS comprises over 70 million patients from 45 health plans. Adult chronic ITP cases were defined as patients with at least two diagnoses (ICD-9 code 287.3) for primary thrombocytopenia separated by at least 6 months between 2002 and 2006, and age 18 or older in 2004. Although ICD-9 code 287.3 is not exclusive to ITP, its sensitivity and specificity are likely to be high (Segal et al., Am J Hem. 2004). A patient’s first identified 287.3 ICD-9 code over the period 2002–2006 had to be in 2004 or earlier and the last one in 2004 or later. Patients had to have continuous enrollment throughout the year 2004. We estimated the 2004 prevalence as the total number of identified chronic ITP cases divided by the total population in the database in 2004 stratified by age group and gender. Estimates of adult cases for the US population were obtained by multiplying the 2004 prevalence estimates by the 2005 adult US census population estimates. The estimated adult prevalence was 23.6 [95% CI 23.4 – 23.8] per 100,000 persons, translating to approximately 52,700 (95% CI 52,200 – 53,100) adult chronic ITP cases in the US. Females had higher prevalence than males (28.1 vs 18.8 per 100,000). In addition, prevalence increased with age (from 17.0 in the age group 18–49 to 36.2 in the age group 65 and older). These results are broadly similar to previously published data and help to better understand the epidemiology of chronic ITP. While the tight CIs predominantly reflect the large data set rather than the precision of the estimates, we believe these data are among the most robust estimates of ITP prevalence and are the first to specifically estimate adult chronic ITP prevalence by age and gender in the US.
Identification of New Alloantigen-Reactive CD8+ Cytotoxic and Suppressor T Cell Subpopulations.
Authors: Bohana-Kashtan Osnat , Levitsky Hyam , Civin Curt I. 
Published: October 2019

We sought to develop a better understanding of the T cells involved in the human allogeneic immune response, in order to eventually engineer a donor graft with reduced GVHD-mediating potential, without ablating general immune competence. Prior studies reported that all the activated CD4+ T cells responding to a specific antigen challenge reside within the CD4high population expressing high levels of membrane CD4. We identified a new population of activated CD8+ T cells that developed during an in vitro allogeneic immune response, along with the allo-activated CD4high T cell population. Analogous to activated CD4+ T cells, this new T cell population was distinguished by up-regulated CD8 (and CD38) expression (CD8highCD38+). In accordance with Martins et al. (Blood 2004, 104:3429), we found that the depletion of the CD4highCD38+ population resulted in reduced 2o response to the original 2nd party stimulators. In contrast, depletion of the CD8highCD38+ population resulted in an increased 2o response to 2nd party cells, with no change in the response to 3rd party or CMV antigens. Elevated numbers of CD8highCD38+ T cells potently reduced the 1o and 2o responses to 2nd party, but not to 3rd party cells or CMV antigens. The complementary, non-activated CD8normalCD38− T cell population had no inhibitory effect. Importantly, we found that
CD8highCD38+ T cells mediated both a specific cytotoxic response (that could be inhibited by the pan-caspase inhibitor, Z-VAD), and a specific suppressive response toward the original 2nd party stimulators (that was not affected by Z-VAD), and within this CD8highCD38+ population, there was a subpopulation of cytotoxic T cells (perforin+LAMP1+CD56+CD11b+CD11c+) and a subpopulation of non-cytotoxic T cells.
Furthermore, we found that although CD8highCD38+ T cells differentially expressed CD28, both CD8highCD38+CD28− and CD8highCD38+CD28− T cells mediated a cytotoxic as well as a suppressor T cell response toward the original 2nd party cells (different from the published suppressive function of CD8+CD28− T cells observed by Liu et al, Int Immunol 1998, 10:775). Upon separation of cytotoxic CD8highCD38+ T cells from suppressor CD8highCD38+ T cells, we will explore the GVHD potential of these 2 novel activated CD8high T cell subpopulations, in a sensitive in vivo xenograft model for GVHD using NOD/SCID/IL2Rγnull immunodeficient mice.
The Use of alpha Interferon To Augment the Graft-Versus-Leukaemia Effect of 2nd Stem Cell Transplants and Donor Lymphocyte Infusions in High Risk Paediatric Leukaemias.
Authors: Cooper Nichola , Amrolia Persis , Goulden Nick , Rao Kanchan , Veys Paul 
Published: October 2019

Abstract
Childhood leukaemia which is refractory to chemotherapy or which relapses early following stem cell transplantation (SCT) has a very poor prognosis, with few patients becoming long-term survivors. Establishing limited graft-versus-host disease (GVHD) in an attempt to exert a graft-versus-leukaemia (GVL) response in these patients remains the only curative option. Second SCT procedures and/or donor lymphocyte infusions (DLI) have achieved some success but the pace of acute leukaemia is often too fast and the occurrence of GVHD/GVL too unpredictable to achieve this goal. We postulated that the addition of alpha interferon (alphaIFN) might induce/augment and control GVHD/GVL in this group of patients and so improve outcome. Thirteen patients underwent initial SCT, for cALL CR2 (n=1), Philadelphia positive ALL CR1 (5), MDS/AML (2), refractory AML (2), and CML (3), with unrelated (9) or sibling donors (4). Eight patients subsequently had a frank haematological relapse and 4 patients a molecular relapse of their leukaemia at a median of 10 months (2–18 months) from 1st SCT. Six patients underwent a 2nd SCT from the same donor, all with reduced intensity conditioned regimes; all received alphaIFN when GVHD did not occur after early withdrawal of immunosuppression. One (ALL CR2) received DLI 2 months after 2nd SCT for mixed chimerism. A further six patients received DLI plus alphaIFN, 3/6 together with low dose DLI (<106/kg); 3/6 had previously received108/kg DLI alone without response. One patient with refractory AML received alphaIFN in conjunction with the initial transplant. The dose of alphaIFN was escalated from 3–5 megaunits/m2 subcutaneously 3–5 days per week as tolerated and continued in the absence of GVHD for up to one year. 3 patients (CML) received concomitant therapy with Glivec, which has now been stopped. Following therapy, 9/13 (69%) patients developed aGVHD: Grade I (1), grade II (4), grade III (4); and 7/11 evaluable patients cGVHD: limited (4), extensive (3). 8 of 13 (62%) remain well and in remission, a median of 23 months (range 12 to 100) from alphaIFN therapy. Five have died: 2 from relapse, one from lung GVHD, one from idiopathic pneumonitis and one from aspergillus infection. Two of 8 surviving patients have ongoing cGVHD requiring immunosuppressive therapy. This data shows promising results for the use of alphaIFN to augment GVL responses in patients with high-risk leukaemia not cured with standard transplant procedures with 62% disease free survival and minimal residual symptoms. Further analysis is warranted.