bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-4 Expression and Constitutive Activation of Erk Signaling In Vivo in Follicular Lymphoma.
Authors: Calvo Katherine R. , Dabir Bhavana , Bandle Russell , Bond Amelia , Jaffe Elaine S. 
Published: October 2019

Abstract
Follicular lymphoma (FL) is characterized by constitutive expression of the Bcl-2 oncoprotein as a consequence of the t(14;18) BCL2/JH translocation in germinal center B-cells. While Bcl-2 confers survival advantage through anti-apoptosis mechanisms, the t(14;18) alone is not sufficient for lymphomagenesis. Evidence suggests that additional factors in the lymph node microenvironment, related to tumor infiltrating T-cells and macrophages, may cooperate with Bcl-2 in orchestrating the malignant phenotype in vivo. In this study we generated proteomic cytokine profiles of patient lymph node tissue lysates from FL (N=53) grades 1–3, and the non-malignant normal counterpart follicular hyperplasia (FH; N=24). Using an ultrasensitive multiplex elisa system we assayed a panel of 11 cytokines known to be secreted by T-cells and/or macrophages including IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IL-12p70, TNF-alpha, and INF gamma. With the exception of IL-4, all cytokines measured showed higher concentrations in FH than FL. In particular, IL-1 beta demonstrated levels 20 times higher in FH; IL-8 levels were 30 times higher; and IL-13 levels were 9 times higher in FH than in FL (p values < 0.005). In contrast, IL-4 levels were 5 times higher in FL than in FH (p < 0.0054). Interestingly, IL-4 levels were highest in FL grade 1 (N=20) in comparison to grade 2 (N=18) and grade 3 (N=12). Immunohistochemical stains for CD3 showed no significant difference in the numbers of CD3+ T-cells in FL samples over FH lymph nodes; or in FL grade 1 vs. grade 2 or grade 3. Hence the differences observed are not due to numerical differences in the total number of T-cells present in FL and FH. We evaluated the activation state of the mitogen activated protein (MAP) kinase signaling pathway, which is activated in response to stimulation by most cytokines in our panel via PI3Kinase. Western blot analysis showed that Erk was constitutively phosphorylated in 14 out of 15 (93%) of FL cases in contrast to only 3 out of 9 (33%) of FH cases. These findings demonstrate that tumor growth in FL is characterized overall by low levels of cytokines within the lymph node microenvironment in comparison to growth associated with the benign counterpart FH. The T-cell secreted cytokine, IL-4, which drives B-cell proliferation, was the only cytokine that showed statistically significant higher expression in the lymph node microenvironment of FL in comparison to FH. Moreover, the intracellular MAP kinase signaling pathway was constitutively activated in vivo in patient FL samples as compared to FH, despite higher overall cytokine levels in FH. These findings suggest that the IL-4 cytokine, and intracellular Erk activation in FL cells, may 1) play a role, in cooperation with Bcl-2, in driving the malignant phenotype in FL and 2) may serve as effective molecular targets for future therapies in FL.
Transferrin Acts as an Iron Sensor Upstream of BMP2b in the Zebrafish.
Authors: Fraenkel Paula G. , Burnett Sarah , Holzheimer Jason , Dooley Kim A. , Wingert Rebecca , Hammerschmidt Matthias , Zon Leonard I. 
Published: October 2019

Abstract
The BMP pathway is important in modulating expression of the iron regulatory hormone hepcidin in response to iron, however how iron levels are sensed is unknown. The zebrafish mutant gavi was identified in a large-scale chemical mutagenesis screen for embryonic anemia. Two alleles were identified: gaviHE067 and gaviIT029, both of which have a homozygous recessive pattern of inheritance. Sequencing of transferrin cDNA from the mutants revealed that each allele of gavi produces an aberrant message and virtually absent transferrin expression. Our previous work demonstrated that hepcidin expression is upregulated following iron dextran injection of the zebrafish embryo. This effect was not impaired by a defect in ferroportin1 (Fraenkel et al., (2005) J Clin Invest115:1532–1541), an iron exporter and ligand for hepcidin. To evaluate the effects of iron loading on transferrin-deficient mutants, gaviIT029 homozygotes at 48 hours post-fertilization were either anesthetized and injected with iron dextran, or anesthetized but not injected. GaviIT029 homozygote embryos exhibited low levels of hepcidin expression that failed to increase following iron injection. Morpholino knockdown of transferrin or transferrin receptor 2 abrogated hepcidin expression, even following iron injection, while knockdown of transferrin receptor 1a (erythroid-specific transferrin receptor 1) or transferrin receptor 1b (a ubiquitously expressed form of transferrin receptor 1) failed to produce this effect. Overexpression of BMP2b, however, rescued hepcidin expression following transferrin knockdown. These findings provide the first evidence that transferrin acts as an in vivo iron sensor upstream of the BMP pathway.
A Randomized Comparison of Total-Marrow Irradiation, Busulfan and Cyclophosphamide with Tandem High-Dose Melphalan in Patients with Multiple Myeloma.
Authors: Knop Stefan , Bauer Katja , Hebart Holger , Wandt Hannes , Trumper Lorenz , Liebisch Peter , Maschmeyer Georg , Peest Dietrich , Wolf Hans H. , Kroger Nikolaus , Straka Christian , Pfreundschuh Michael , Coser Paolo , Meisner Christoph , Kanz Lothar , Einsele Hermann 
Published: October 2019

Abstract
Background Myeloablative chemotherapy with support of autologous peripheral blood stem cells (APBSC) has widely been accepted as a standard of care in patients (pts) with newly diagnosed multiple myeloma (MM). High-dose (HD) melphalan (Mel) 200 mg/m2 was considered superior to total-body irradiaton (TBI) plus Mel 140 mg/m2 for toxicity reasons. Since MM plasma cells are inherently responsive to irradiation, our group evaluated TBI aimed at reduced organ toxicity by shielding lungs and liver (total-marrow irradiation [TMI], 9 Gy) combined with busulfan (Bu, 12 mg/kg) and cyclophosphamide (Cy, 120 mg/kg) in a previous phase I/II trial (Einsele et al, Bone Marrow Transplant, 2003).
Patients and methods In the current study (DSMM I), subjects with previously untreated MM in Durie-Salmon stages II/III were randomly assigned to either receive one course of TMI/Bu/Cy versus two cycles of HD Mel 200 mg/m2 each with APBSC transplantation if having had an adequate number of stem cells collected and at least stable disease. Primary end point was event-free survival (EFS), secondary end points overall (OS) and disease-free survival (DFS).
Results A total of 294 pts (median age, 54 years), 246 of whom completed stem cell harvest were enrolled between 8/1998 and 1/2002 by 46 centres. Eventually, 198 (n=100 TMI/Bu/Cy and n=98 HD Mel) pts were randomized and included into the ITT population. The safety population (n=80 TMI and n=118 HD Mel, due to 18 pts switching to Mel) was analyzed for toxicity and response. CR rate before HD therapy was 7.0% (7/100) in the TMI and 6.1% (6/98) in the Mel arm respectively. Significantly more pts receiving TMI/Bu/Cy experienced WHO grades 3 and 4 pulmonary and gastrointestinal toxicity and pain. Following HDT, CR rate increased to 17.5% (14/80, TMI) and 32.2% (38/118, HD Mel; p=.022) respectively. After a median follow-up of 1447 days, median EFS in the TMI group was 1161 days versus 1090 days for HD Mel (p=.812). Probability of 4-year OS was 72.7% (95%-CI: 62.1–80.7) with TMI and also 72.7% (95%-CI: 61.7–81.1) after HD Mel (p=.754). For pts in CR following HD therapy, probability of 4 year DFS was 62.4% (95%-CI: 33.6–81.6) for TMI vs. 50.4% (95%-CI: 30.6–67.3) for HD-Mel (p=.138).
Conclusion In this randomized trial on pts with newly diagnosed MM, the irradiation-based regimen was associated with more pulmonary and GI toxicity when compared to HD Mel. Incidences of other toxicities including hepatotoxicity, however, were not different between the two treatment arms. CR rate was superior for HD-Mel, while there was no difference in OS and EFS between the two treatment arms. Subjects achieving CR may be more likely to enjoy prolonged DFS after TMI/Bu/Cy.
Clinical Relevance of a Pharmacogenetic Approach Using Multiple Candidate Gene Polymorphisms To Predict Response and Resistance to Imatinib Mesylate Therapy in Chronic Myeloid Leukemia.
Authors: Kim Dong Hwan (Dennis) , Sriharsha Lakshmi , Xu Wei , Kamel-Reid Suzanne , Liu Xiangdong , Siminovitch Katherine , Messner Hans , Lipton Jeffrey 
Published: October 2019

Abstract
Background: Imatinib resistance (IR) is a well known cause of treatment failure in chronic myeloid leukemia (CML) patients being treated with imatinib mesylate (IM). Several cellular and genetic mechanisms of IR have been proposed including amplification and overexpression of the BCR-ABL gene, the presence of specific point mutations and MDR1 gene overexpression.
Methods: We investigated the impact of 16 single nucleotide polymorphisms (SNPs) in 5 genes potentially associated with pharmacogenetics of IM (ABCB1, multidrug resistance 1; ABCG2, breast-cancer resistance protein; CYP3A5, cytochrome P450 3A5; HOCT1, human organic cation transporter 1; AGP1, alpha-1-acid glycoprotein-1, plasma protein binding to IM). The major endpoints included:
response to IM: cytogenetic (CyR) or molecular response (MoR); resistance to IM: loss of response (LOR), treatment-failure (including primary resistance or LOR); progression to accelerated phase (AP) or blast crisis (BC), or death; and need for IM dose escalation to overcome resistance or LOR. The DNAs from peripheral blood samples were genotyped using the Sequenom MassARRAY system based on MALDI-TOF technique.
The study population included 229 patients whose clinical outcomes following IM therapy were evaluated from January 2000 to January 2007 (male:female 96:133; median age at start of IM, 53 years-old; white:non-white 170:59; chronic phase:AP:BC, 199:23:3).
Results: The frequencies of genotypes in 16 SNPs are summarized in Table 1. The GG allele in ABCG2 (rs2231137), AA allele in CYP3A5 (rs776746) and advanced stage were significantly associated with poor response to IM, while GG allele at HOCT1 (rs683369) and advanced stage correlated with high rate of LOR or treatment failure. The CC allele in ABCG2 (rs2231142) was also identified as an independent predictor of more frequent need for IM dose escalation. The results of multivariate analyses are summarized in Table 2.
Conclusion: Using a novel, multiple candidate gene approach based on the pharmacogenetics of Imatinib mesylate, we identified several SNP candidates in patients with CML that are potential predictors of clinical response to IM (rs2231137, ABCG2 or rs776746, CYP3A5), resistance to IM (rs683369, HOCT1) and need for IM dose escalation (rs2231142, ABCG2). Further studies are warranted to validate the role of these SNPs in the early identification of individuals with CML who may not respond optimally to standard IM therapy.
Table 1. The genotype frequency and clinical significance in 16 SNPs Table 2. Multivariate regression model based on Cox's proportional hazard model for each-point
Th-17 Lymphocytes, Not Regulatory T Cells, Mediate Immune Regulation of Bone Marrow Infiltrating Lymphocytes and Induce Osteoclast Activation in Multiple Myeloma.
Authors: Noonan Kimberly A. , Anderson Judy , Mgebroff Stephanie , Serafini Paolo , Roodman David , Borrello Ivan 
Published: October 2019

Abstract
The bone marrow of myeloma patients is a reservoir of marrow infiltrating lymphocytes (MILs) that upon activation with anti-CD3 and anti-CD28 antibodies show significant tumor specificity against both mature myeloma plasma cells and their clonotypic precursors, whereas activated peripheral blood lymphocytes (PBLs) fail to demonstrate any tumor specificity. Here we examine the major differences between MILs and PBLs of both myeloma patients and normal donors. In myeloma, CD4+/CD25+ MILs lack a regulatory T-cell (Treg) phenotype as shown by the absence of FoxP3 expression and the inability to suppress tumor-specific proliferation of effector T cells. In contrast, the CD4+/CD25+ PBLs express FoxP3 and suppress tumor specific proliferation. Analysis of MILs and PBLs from normal individuals reveals the exact opposite (more Tregs in the marrow than blood). Considering the abundance of plasma cell-derived IL-6 in myeloma marrow, we hypothesize that IL-6 skewing of MILs towards the effector Th17 phenotype could explain the reciprocal paucity of Tregs in myeloma and relative abundance in normal bone marrows. Intracellular staining shows increased expression of IL-17 in myeloma CD4 MILs (2.7%) as compared to either myeloma PBLs (0.02%) or normal MILs (0.7%). The Th17 cytokine secretion pattern also appears more pronounced in myeloma-derived MILs as compared to either myeloma PBLs or normal MILs respectively (see table). Th17 T cells have recently been implicated in osteoclastogenesis of rheumatoid arthritis. In fact, bone marrow serum IL-17 levels correlate with the degree of lytic bone disease. In experiments utilizing M-CSF and RANK-L as the osteoclast differentiation medium, we show that the addition of IL-17 to the culture medium increases the number of mature osteoclasts by 2.3 fold whereas γIFN reduces mature osteoclast numbers by 80%. These findings explain the profound differences seen in MILs of normal and myeloma patients and implicate Th17 MILs in the generation of lytic bone disease. Activation of myeloma MILs with anti-CD3/CD28 skews the population from Th17 to a γIFN-producing Th1 phenotype. As such, targeting Th17 T cells or utilizing adoptive T cell therapy with activated, γIFN-producing MILs may represent a novel therapeutic mechanism to target osteoclastogenesis and reduce bone disease in myeloma.
Th-17 Cytokine Profile N=56 MM MILs MM PBLs NL MILs IL-6 (pg/ml) 30 3.7 6.5 IL-23 (pg/ml) 246 19.6 35.9 IL-17 (pg/ml) 12.2 0.4 5.1
Isoprostane (8-Epiprostaglandin F2α) Plasma Concentration and Aspirin Resistance in Patients after Ischaemic Stroke.
Authors: Zawilska Krystyna M. , Zytkiewicz Marcin M. , Hanszke Ewelina E. , Turowiecka Zofia Z. , Gielwanowska Liwia L. , Psuja Piotr P. 
Published: October 2019

Abstract
Background The limited efficacy of secondary prevention for ischaemic stroke may be partially related to aspirin resistance leading to continuous generation of intraplatelet thromboxane A2. Besides other underlying metabolic mechanisms, an oxidant stress along with nonenzymatic biosynthesis of isoprostanes supporting the platelet activation has been suggested. Aims We analyzed the incidence of aspirin resistance in survivors of ischaemic stroke and compared the usefulness of some platelet tests designed for its laboratory exploration. Also we searched for relationship between isoprostane concentration and results of platelet function tests.
Material and Methods Forty four patients, at least a month after acute onset of ischaemic stroke were included into the study. All of them have been receiving 75–150 mg aspirin daily at least for a month. The control group for 11-dehydro Thromboxane B2 measurements consisted of 12 healthy volunteers. The platelet function was investigated by platelet aggregation tests induced by either ADP (3.5 and 5.0 μM), collagen (2 μg/ml) or arachidonic acid (AA) (0.6 mM) and by measurement of closure time in PFA-100 analyzer. Thromboxane A2 metabolite - 11-dehydro Thromboxane B2 (11-dTxB2) and isoprostane - 8-epi Prostaglandin F2α (8-epiPgF2α) plasma concentration by immunoenzymatic method (EIA Kits from Cayman Chemicals) were also determined. The aspirin ingestion was controlled by diminished intraplatelet concentration of malonyldialdehyde. Aspirin resistance has been determined by the following criteria: the intensity of platelet aggregation induced by ADP >60%, collagen >70%, AA >20%, PFA-100 closure time <165 s and 11-dTxB2 concentration >mean of the control group minus SD.
Results Table 1 Statistically significant inverse correlations have been found between the plasma concentration of 11-dTxB2 and PFA-100 (Col/Epi) closure time (r= −0.31; p= 0.039) as well as between plasma concentration of 8-epiPgF2α and PFA-100 (Col/Epi) closure time (r= −0.36; p= 0.019).
Summary/conclusions
Laboratory platelet function tests reveal aspirin resistance in almost half of patients after ischaemic stroke. The most significant correlation has been found between plasma concentration of 11-dehydro Thromboxane B2 and PFA-100 closure time. An important interrelationship observed between PFA-100 closure time and plasma concentration of 8-epi Prostaglandin F2α may support the hypothesis of nonenzymatic production of isoprostanoids with platelet proaggregatory activity, playing a role in aspirin resistance.
The frequency of aspirin resistance in patients after ischaemic stroke ADP 3.5μM ADP 5.0μM Collagen Arachidonic acid PFA-100 11-dTxB2 45% 52% 20% 7% 52% 43%
Sequential VAD (Vincristine, Adriamycin, Dexamethasone) and VTD (VELCADE®, Thalidomide, Dexamethasone) Induction Followed by High-Dose Therapy with Autologous Stem Cell Transplantation and Maintenance Treatment with VELCADE for Newly Diagnosed Multiple Myeloma: Interim Results of Phase II Trial.
Authors: Yoon Sung-Soo , Kim Hye Jin , Lee Dong Soon , Eom Hyeon Seok , Jang Jun Ho , Chung Ju Seup , Kang Hye Jin , Kim Cheol Soo , Kim Kihyun , Lee Jae Hoon , Lee Sang Jae , Min Yu Hong , Seong Chu Myong , Sohn Sang Kyun , Suh Cheolwon , Won Jong Ho 
Published: October 2019

Abstract
Introduction Effective reduction of myeloma before autologous stem cell transplantation (ASCT) prolongs survival in multiple myeloma patients. Recently, incorporation of novel agents resulted in improved response rate and reduced side effect in newly diagnosed multiple myeloma.
Method: Patients are planned to receive 2 cycles of VAD (vincristine 0.4mg D1-4, adriamycin 9mg/m2 D1-4, dexamethasone 40mg D1-4, 9–12 every 3 weeks), and VTD (bortezomib 1.3mg/m2 D1, 4, 8, 11, thalidomide 100mg daily, dexamethasone 40mg D1-4, 9–12 every 3 weeks). High dose melphalan (200mg/m2) is used as a conditioning regimen for ASCT. Bortezomib (1.3mg/m2) as a maintenance treatment is administered weekly x 4 times every 6 weeks for 4 cycles after ASCT. Response was assessed by EBMT criteria, with additional category of nCR. Adverse events were graded by the NCI-CTCAE, Version 3.0.
Result: At this interim analysis, 60 patients have been entered into the ongoing trial, and efficacy could be assessed in 53 patients. After 2 cycles of VAD, response rate was 70%. After VTD, two patients showed further improvement with additional CR, and an overall response was 97% with 14% CR. Especially, patients with poor prognostic cytogenetics (n=6) all responded after VTD. So far, autologous stem cells were successfully collected in all 28 patients with a median CD34+ count of 7.8 x 106/kg (range, 2.17–44.7 x 106/kg). In 24 patients who underwent autologous stem cell transplantation, five patients gained additional CR. There was no progression in patients completed bortezomib maintenance (n=9, CR 77%). The median follow-up duration was 6 months, median time to response was 1.4 months, and median overall survival was not reached. Grade 3,4 hematologic toxicity was more frequently observed after VAD than VTD (anemia 15.8%, 4.6%, neutropenia 7.9%, 3.5%), and incidence of grade 2,3 peripheral neuropathy was low (VAD 3.5%, VTD 7%).
Conclusion: Sequential VAD and VTD induction therapy in newly diagnosed multiple myeloma was highly effective, even in patients with poor prognostic cytogenetics, and did not prejudice stem cell collection. VTD could have contributed to increased RR and minimized side effects. An updated results will be presented at the ASH meeting.
*Protocol Number: KMM51-NCT00378755.
Anemia and Iron Deficiency in Strenuously Trained Adolescents.
Authors: Merkel Drorit , Huerta Michael , Grotto Itamar , Rachmilewitz Eliezer A. , Fibach Eitan , Epstein Yoram , Shpilberg Ofer 
Published: October 2019

Abstract
Background: Healthy soldiers serving in combat units have a higher prevalence of anemia than age- and sex-matched civilians. This may be a “pseudo-anemia” caused by the hemodilution typical among training athletes, or a “true anemia” due to reduced total body iron stores.
Objective: To investigate the incidence of iron-deficiency anemia in recruits to the Israel Defence Forces during their first 6 months of intense combat training. This is a follow-up study to previous publication of measured values on induction.
Methods: Blood was collected from new recruits to an elite infantry unit before training. After 6 months, 153 paired samples were collected from the initial group. Total blood count and serum iron, transferrin and ferritin were measured at both time points. Soluble transferrin receptor (sTfR) was measured in 119 of the paired samples, and sTfR/log ferritin ratio was calculated.
Results: At recruitment, mean hemoglobin concentration was 14.7±0.9 g/dL (range 11.5–16.8). Iron-transferrin saturation was 34.1±13.6%, and mean ferritin concentration was 53.6±33.2 ng/mL. Twenty-seven participants (17.6%) were anemic (Hb<14g/dL), and 14.9% were iron-deficient (ferritin level <22 mg/dL). At the end of the follow-up period, 50.3% of the soldiers examined were anemic, and 27.3% had signs of iron-store depletion. Analysis of the paired samples showed an average reduction of 0.83 g/dL in hemoglobin level, and of 9.8mg/dL in ferritin levels (p<0.001 for both). sTfR increased slightly from 1.9 to 2.1mg/dL (p<0.003) among the recruits who became anemic during the follow-up period.
Conclusion: Nearly half the new recruits studied endure mild anemia after the first 6 months of training. Iron store depletion was observed among 24.5% of the cohort after 6 months, as opposed to 15% at recruitment. Overall, these changes were not accompanied by a significant increase in sTfR, but among the subset of anemic soldiers, there was a slight increase in this index. From the iron status analyses it can be concluded that in half the cases, the observed new-onset anemia was attributable to iron deficiency, and in the remainder, to hemodilution. The high incidence of iron deficiency in young healthy recruits is an important issue. The therapeutic implications of these findings require further evaluation.