bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autologous Stem Cell Transplantation and Rituximab for Mantle Cell Lymphoma.
Authors: Capote Francisco Javier , González-Barca E. , Bergua J.M. , Pascual M.J. , García-Boyero R. , Ribera J.M. , Jiménez A. , Romero A. , Giraldo P. , Fernández-Calvo J. , Ramírez J. , Amigo M.L. , Muñoz M.I. , Caballero D. , Guinot M. , Oriol A. , Queizán J.A. , Cantalapiedra A. , Moraleda J.M. , Grande C. , Palomera L. , Almagro M. , Leon A. , Fernández-Sevilla A. , Gil J.L. 
Published: October 2019

Abstract
Introduction: Mantle cell lymphoma (MCL) is a mature B-cell lymphoma comprising up 5% of non-Hodgkins lymphomas. Although the prognosis for MCL patients has improved in recent years, the outlook for those with advanced or recurrent disease remains poor and the role of hematopoietic stem cell transplantation is unclear. The HyperCVAD-M/A regimen (fractionated high-dose cyclophosphamide, vincristine, doxorubicin and prednisolone alternated with methotraxate and cytarabine) has yielded encouraging results when combined with autologous stem cell transplantation (ASCT). In an effort to improve these results further, we have combined rituximab in vivo purging and post-transplant consolidation with HyperCVAD-M/A plus ASCT.
Methods: Patients aged <65 years with previously untreated or relapsed MCL were treated with four courses of HyperCVAD-M/A followed by four once-weekly doses of rituximab 375mg/m2 as purging prior to stem cell mobilization and harvesting, high-dose chemotherapy (ICT-CY or BEAM), stem cell reinfusion and four further doses of rituximab immunotherapy post-transplant.
Results: Of the 40 patients enrolled so far, 20 (15 male, 5 female; 18 previously untreated) have been transplanted. The median age was 50 years (range 38–63 years). After the final post-ASCT immunotherapy all 20 patients were in clinical complete remission. With a median follow-up of 36 months from diagnosis (range 7–64 months), 18 patients remain alive with 13 in complete remission. One patient died 15 months post-ASCT without evidence of disease recurrence. Kaplan-Meier estimates of 5-year overall and event-free survival are 90% and 65% respectively.
Conclusions: This approach seems safe and feasible and produces durable remissions; longer follow-up of a more patients will be required to assess the effect of the procedure on survival.
Nucleotide Insertions and Deletions in Chronic Lymphocytic Leukemia. A CLL Specific Deletion among IGHV3-21 Expressing Cases with Stereotyped Receptors.
Authors: Belessi Chrysoula , Davi Frédéric , Stamatopoulos Kostas , Degano Massimo , Andreou Thanassis , Moreno Carol , Merle-Beral Hélène , Crespo Marta , Laoutaris Nikolaos , Montserrat Emili , Caligaris-Cappio Federico , Anagnostopoulos Achilles , Ghia Paolo 
Published: October 2019

Abstract
Insertions/duplications and deletions (I/D/Ds) in IG variable region genes are infrequent sequelae of the somatic hypermutation process. In the present study, we document the occurrence of nucleotide I/D/Ds in rearranged IGHV genes in CLL patients. Among 809 IGHV-D-J sequences amplified in 760 CLL cases, 9 (1.11%) and 15 (1.85%) sequences exhibited IGHV sequence changes consistent with nucleotide insertions/duplications or deletions, respectively. I/D/Ds were found in genes of the IGHV1, IGHV3 and IGHV4 subgroups, always within mutated IGHV-D-J rearrangements. In 21/24 cases, the inserted/duplicated or lost nucleotides occurred in multiples of 3; therefore, the original reading frame was maintained and a potentially intact receptor was coded. I/D/Ds were located completely or in part within CDRs in 21/24 cases; sequence motifs (AGY, AGA, AAC trinucleotides) that resemble intrinsic hotspots for somatic hypermutation were identified in 21/24 sequences. Short stretches with high homology (misalignment feet) that would offer the DNA polymerase an alternative template for re-annealing in cases of replication slippage were identified in all CLL sequences carrying I/D/Ds. I/D/Ds were generated somatically since (i) they always occurred in cases with somatic mutations, (ii) they could not be found on sequencing analysis of the corresponding germline IGHV genes amplified on DNA isolated from selected patients’ neutrophils, thus excluding a possible genetic polymorphism. The incidence of I/D/Ds in CLL is consistent with previous reports in normal, autoreactive and neoplastic human B cells, thus seemingly indicating that these modifications generally arise without any particular disease-specific associations. A striking exception to this rule was identified in the case of CLL IGHV3-21 expressing cases: one aminoacid was deleted from the CDR2 region in 16/74 (21.6%) IGHV3-21 CLL sequences (database-derived IGHV3-21 CLL cases + present series) vs. only 2/340 (0.59%) non-CLL IGHV3-21 sequences; 15/16 CLL IGHV3-21 sequences carrying this deletion belonged to a subset with unique, shared HCDR3s and light chain CDR3 motifs. The effect of the CDR2 deletion on the structure of the IG molecules in this subset of CLL patients was studied with molecular modelling and dynamics simulations. The models suggested that the deletion could be accommodated without significantly affecting the local structure. The close association of deletions in CDR2 with the homologous subset of IGHV3-21 expressing CLL cases provides further evidence for the importance of an antigenic drive in malignant transformation and/or maintenance of the neoplastic clone in at least some CLL cases.
Interaction between B-CLL Cells and Lymphoid Microenvironment Leads to Activation of PI3-K/Akt Pathway and Inhibition of Apoptosis.
Authors: Shehata Medhat , Schnabl Susanne , Demirtas Dita , Schwarzmeier Josef D. , Hilgarth Martin , Duechler Markus , Hubmann Rainer , Jaeger Ulrich 
Published: October 2019

Abstract
Inhibition of apoptosis and long survival leads to accumulation of the leukemic cells in B cell chronic lymphocytic leukemia (B-CLL). This could be due to activation of anti-apoptotic cascades in CLL cells through interaction with their lymphoid microenvironment. Therefore, we investigated the role of tumor microenvironment in prolongation of survival of B-CLL cells and activation of the potent anti-apoptotic PI3-K/Akt pathway.
Stromal fibroblasts of bone marrow (BMFs), spleen (SF) and lymph gland (LGF) were used as an in vitro model for lymphoid microenvironment and we tested their ability to inhibit spontaneous apoptosis of B-CLL cells. Co-culture of B-CLL cells with human BMFs, LGF, and SF significantly inhibited apoptosis and prolonged survival of the leukemic cells in comparison to suspension cultures and to co-cultures with fibroblasts obtained from non-lymphoid organs. Trans-well culture experiments indicated that cell-cell interaction and soluble mediators are essential for this supportive effect. To explore the involvement of PI3-K/Akt pathway in the anti-apoptotic effect of stromal fibroblasts, co-cultures were performed in presence of PI3-K inhibitors (wortmannin or Ly294002) or siRNAs against PI3-K (p110ß subunit) and Akt1. These inhibitors significantly reduced the supportive effect of stromal fibroblasts and induced apoptosis in B-CLL cells. Interestingly, the leukemic cells were far more sensitive to PI3-K inhibition than T cells, monocytes and fibroblasts. Induction of apoptosis was associated with a significant decrease in the intracellular PIP3, PI3-K, PDK1 and Akt1, NF-kappa B, IKK and de-phosphorylation/activation of tumor suppressor protein PTEN. Studies using phosphospecific anti-PTEN antibody demonstrated that PBMC of CLL patients (n=40) highly express a phosphorylated form of PTEN. The results demonstrate that the PI3-K/Akt pathway is involved in inhibition of apoptosis of B-CLL cells and suggest that interaction of the leukemic cells with lymphoid microenvironment maintains the activation of this pathway. The data also suggest that targeting this pathway represents a new option for designing novel therapeutic strategies in B-CLL.
Hypoxia Inducible Factor-1α Is over Expressed in CLL B Cells Because of an Impaired Proteasome Pathway Associated with Defective Interaction with von Hippel-Landau Protein.
Authors: Lee Yean K. , Strege Ann K. , Bone Nancy D. , Wellik Linda E. , Chan D. A. , Giaccia A. J. , Mukhopadhyay Debabrata , Kay Neil E. 
Published: October 2019

Abstract
We have found that CLL B cells spontaneously secrete vascular endothelial growth factor (VEGF) and that a VEGF autocrine pathway can induce apoptosis resistance in these cells. Recently, we also found that hypoxia-inducible factor-1 alpha (HIF-1α) is highly expressed in CLL B cells. Since this protein is a potent transcription factor for the induction of VEGF, we were interested in further definition of HIF-1α regulation and its function in CLL B cells. CLL blood B cells overexpress HIF-1α protein but not mRNA for HIF-1α compared to normal blood and splenic B cells. Immunohistochemistry (IHC) showed that circulating blood CLL B cells and a subset of CLL marrow cells uniformly express HIF. Hypoxic conditions (i.e., 1% O2) did not increase the protein levels of HIF-1α nor mRNA for HIF-1α in CLL B cells, indicating that the high HIF-1α protein level is due to post-translation modification. Blockade of signaling pathways known to increase HIF-1α levels also did not alter the high levels of HIF-1α in CLL B cells. IHC and nuclear extraction assay demonstrated that HIF-1α was predominantly located in the CLL B cell nucleus. In addition, the nuclear extract when immunoprecipitated for HIF-1α was shown to be complexed with the co-activator p300, indicating that HIF-1α is transcriptionally active. Co-immunoprecipitation assay showed that HIF-1α from CLL B cells does not associate and form a complex with von Hippel-Landau protein tumor suppressor (pVHL), indicating that the proteasome dependent degradation pathway for HIF-1α protein in CLL B cells is dysfunctional. Using immunoblot or IHC methods, we were unable to detect pVHL protein in CLL B cells; however, we were able to use immunoprecipitation of CLL B cell lysates to demonstrate there is pVHL in CLL B cells. Prolyl hydroxylases (PHD 1, 2, and 3) are negative regulators for HIF-1α via hydroxylation of amino acid prolines in the oxygen degradation domain (ODD) which permits interaction with pVHL. RT-PCR results revealed that there is a subset of CLL patients who had ≥ 50% reduction of PHD 1 and 3 mRNA levels. However using a hydroxylation specific polyclonal antibody we found that HIF-1α from CLL B cells is indeed hydroxylated. Finally, silencing of HIF-1α by RNA interference in CLL B cells was associated with a selective decrease in VEGF mRNA levels but not VEGF-R1, Mcl-1 and prolyl hydroxylases (PHD 1–3) other downstream target genes of HIF-1α. These data show that the high endogenous HIF-1α levels in CLL B cells are due to a defect in HIF-1α degradation via the proteosomal pathway. We believe that this abnormality is linked to the autocrine VEGF pathway in CLL B cells and ultimately results in increases in their apoptotic resistance. Inhibition of HIF-1α levels may be of therapeutic benefit to CLL patients.
FCR-3 as Frontline Therapy for Patients with Chronic Lymphocytic Leukemia (CLL).
Authors: O’Brien Susan , Wierda William G. , Faderl Stefan , Ferrajoli Alessandra , Bueso-Ramos Carlos E. , Browning Mary , Kantarjian Hagop M. , Keating Michael J. 
Published: October 2019

Abstract
The combination regimen of Fludarabine, Cyclophosphamide, and Rituximab (FCR) has produced high overall response (OR) rates and complete remission (CR) rates in patients receiving this as initial treatment for CLL. Earlier studies have shown that dose-dense or dose-intensified regimens of single-agent Rituximab produced higher response rates in patients with CLL. The current regimen, FCR-3, was based on the hypothesis that increasing the amount of Rituximab in the combination might improve response rates and remission duration. Doses of chemotherapy were Fludarabine 25 mg/m2/Dx3, and Cyclophosphamide 250 mg/m2/Dx3 given monthly. Rituximab was given at 375 mg/m2 as the first dose and 500 mg/m2 for all subsequent doses. On each day of the chemotherapy a dose of Rituximab was given so 3 doses were given monthly. With the previous FCR regimen 45% of patients achieved a CR or nPR and had <5% CD19+ 5 positive cells in the marrow at the completion of 3 cycles of therapy (rapid response, RR). The median time to progression (TTP) in that group has not been reached; it was 3 years in patients who did not achieve that endpoint (p<.001). The current protocol aimed to increase that response rate from 45% to 60%. Sixty-five patients were treated. Eighty percent were men. The median age was 59 years (27–82). Rai Stage 3–4 disease was present in 25% of patients. Median WBC count was 92.4 x 103/ul (7.9–363). Median B2-microglobulin was 3.8 (1.6–10.1). Unfavorable FISH abnormalities were present in 35% of 52 evaluable patients. Somatic hypermutation status was available for 44 patients; 61% were unmutated. ZAP-70 expression analysis performed by immunostaining or flow cytometry was positive in 62% of 47 evaluable patients. Results of FCR-3 in comparison to FCR are shown in the table.
3 Cycles 6 Cycles No. RR(%) OR CR Flow<5% FCR 300 45 95 72 82 FCR-3 65 45 94 65 74
PCR negativity using consensus primer PCR was achieved in 49% of patients at the end of therapy. Median number of days between courses ranges from 29–35 per course (overall range 27–95). Eighty-five percent of patients completed 6 cycles. No patient has progressed with a median follow-up of 10 months. With limited follow-up the addition of 3 doses of Rituximab to FC chemotherapy does not appear to provide greater benefit than one dose.
The Effect of Melatonin on Thrombopoiesis in Irradiated Mice.
Authors: Yang Mo , Li Karen Kwai Har , Huang Wei Zhe , Li Xiao Jing , Pong Nga Hin , Liu Ai Guo , Li Chi Kong , Fok Tai Fai , Ng Pak Cheung 
Published: October 2019

Abstract
We have reported that serotonin (5-hydroxytryptamine, 5-HT) significantly stimulates megakaryocytopoiesis. It has been known that platelet count can be increased in stress conditions and serotonin has an effect on actin re-organization and promotes nitric oxide (NO) synthesis via 5-HT2A receptors. NO may induce apoptosis of mature megakaryocytes to produce platelets. Taken together, we proposed a possible mechanism of serotonin on megakaryocyte differentiation and platelet formation: serotonin enhances the differentiation and maturation of megakaryocytes via 5-HT2BR and actin re-organization; and serotonin is involved in the process of platelet formation by promoting the synthesis of NO to induce cell apoptosis via 5-HT2AR. Serotonin is the precursor of melatonin. The effect of melatonin on thrombopoiesis in irradiated mice (4 Gy) was investigated in this study. Eighteen mice were randomly divided into three groups (6 in each). Group 1 (normal control, N) received no irradiation or melatonin; Group 2 (model control, C) and Group 3 (melatonin, M) received 4 Gy total body irradiation for 3 minutes. After irradiation, melatonin (10mg/kg.d) was injected intraperitoneally into Group M for consecutively 21 days. And Group C was administered normal saline instead of melatonin. Peripheral blood platelets, white blood cells (WBC), and red blood cells (RBC) were analyzed from the three groups on day 0, 7, 14, and 21. All the mice were executed to collect bone marrow cells for the assay of CFU-MK and CFU-F (fibroblastoid). Our results showed that melatonin enhanced the recovery of platelets and WBC counts. Moreover, melatonin also promoted CFU-MK (20 ± 3 vs 11 ± 2 colonies/2 x105 cells) and CFU-F formation (28 ± 10 vs 15 ± 3 colonies/2 x106 cells). We also investigated the in-vitro effect of melatonin on CFU-MK formation at different doses (0–500 nM). The results showed that melatonin significantly promoted CFU-MK formation at a dose dependant manner. The maximal concentration was at 200 nM (P<0.01, N=6) compared with the control. The size of CFU-MK with melatonin treatment was much larger and each MK cell was more mature. Our studies showed that melatonin had a promoting effect on thrombopoiesis in this model and also stimulated in vitro CFU-MK formation. Thus, melatonin could be developed as an alternative drug for the treatment of thrombocytopenia.
Tissue Factor-Driven Enhanced Platelet Aggregation in Flowing Blood in a Rabbit Model of Childhood Hemolytic Uremic Syndrome.
Authors: Grabowski Eric F. , Nestoridi Eirini , Garcia Alexander , Fox James G. , Ingelfinger Julie R. 
Published: October 2019

Abstract
The Dutch belted rabbit constitutes a promising animal model of the childhood hemolytic uremic syndrome (HUS; A. Garcia, J. Infect. Dis. 2002). When Shiga toxin (Stx)-producing E. coli are administered orally, these rabbits develop HUS. Platelet-fibrin thrombus formation in this model was examined on unfixed renal tissue (5 μm sections) after mounting on 150-μm glass slides in test animal-control animal pairs. Normal donor blood, collected into 4U/ml low MW heparin and 10 μM quinacrine dihydrochoride, was drawn at shear rates of 270–650 sec-1 through a parallel-plate flow chamber for which one surface was one of the above slides. Such shear rates give rise to shear stresses which approach the 20–25 dynes/cm2 estimated to exist in glomerular arterioles. Platelets depositing on the sections were imaged in real time using epifluorescence digital videomicroscopy, with quantitation of adherent platelets via image processing. For 16 pairs of test and control sections, the percent of image pixels occupied by adherent platelets after 6–9 min of blood flow was 17.7 ± 3.06 (mean ± SD) for test sections vs 5.71 ± 5.18 for controls (P<0.001). This 3.1-fold increase for sections from Stx-treated rabbits is all the more striking considering that individual platelet aggregates on these sections were also larger and thicker, as judged from greater local pixel intensities. In 4 additional experiments, preincubation of sections with 300 nM of a monoclonal antibody (courtesy of Y. Nemerson, Mt. Sinai Medical Center, NY, NY) directed against human tissue factor (TF) with known cross-reactivity to rabbit TF markedly reduced platelet adhesion/aggregation on both test and control sections. Residual platelet deposition, which only slightly favored test sections, may be due to either a non-TF-dependent mechanism or incomplete neutralization of TF activity by the amount of antibody used. Studies of parallel fibrin deposition are in progress. Thus, renal tissue from the Dutch belted HUS model is reactive to platelets in large part via expression of TF, an expression augmented on test sections. These observations support the hypothesis that the TF pathway of coagulation plays a major role in childhood HUS.
Does the Platelet Factor 4 (PF4) ELISA Correlate with the Clinical Diagnosis of Type II Heparin-Induced Thrombocytopenia?.
Authors: Streiff Michael B. , Kickler Thomas S. , Weir Edward G. 
Published: October 2019

Type II heparin-induced thrombocytopenia (HIT) is an antibody-mediated complication of heparin therapy that is associated with a consumptive thrombocytopenia and a high risk of thromboembolism. It is now known that antibodies associated with type II HIT recognize sites on PF4 when complexed with heparin. Due to technical challenges associated with the serotonin release assay, most clinicians rely on the PF4 ELISA to establish a laboratory diagnosis of HIT. However, it is unclear whether the ELISA has predictive value in identifying those patients who have clinically apparent disease and whether ELISA optical density (OD) measurements correlate with the development of thrombosis. We retrospectively reviewed the medical records of 103 sequential patients who tested positive for HIT by a commercial PF4 ELISA from 2000–2002. While blinded to ELISA OD values, we recorded patient age and gender, admission diagnosis, hospital course, type of heparin administered, route of administration, baseline and nadir platelet counts, timing of thrombocytopenia, HIT therapy, clinical thrombotic events within 30 days, and other potential causes of thrombocytopenia to determine the clinical likelihood of HIT. The association between OD values and both the clinical likelihood of HIT and objectively confirmed thrombotic events was analyzed using the Pearson Chi-square test. Median patient age was 62 yrs and 52% of patients were female. Those with cardiac disease, cancer, and venous thromboembolism constituted 44%, 19% and 9% of the study population, respectively. Median platelet count nadir was 40K/mL (range, 2K–90K), and average onset of thrombocytopenia was 6.5 days. Median ELISA OD value was 0.77 (0.40–3.81). The clinical likelihood of HIT was noted in 24/103(23%) patients, and 22(21%) suffered a thrombotic event. An ELISA OD >1.0 was significantly associated with the clinical likelihood of HIT (p=0.03), though OD values ranged from 0.41 to 3.81 among these patients. Neither an OD threshold of 1.0 or 1.5 was associated with thromboembolism (p=0.2 and p=0.1, respectively). In contrast, the clinical likelihood of HIT was significantly associated with subsequent thrombosis (p=0.002). ELISA results were not significantly associated with the extent or onset of thrombocytopenia, type of heparin or route administered, patient age or gender, admission diagnosis, or hospital course. In conclusion, PF4 ELISA OD values are significantly associated with the clinical likelihood of HIT but are not predictive of HIT on an individual patient basis. Clinical judgment but not ELISA OD values is associated with subsequent thromboembolism. In summary, the clinical likelihood of HIT, and not ELISA results, remains the best parameter by which to modify anticoagulation therapy in patients being assessed for HIT.
Figure Figure
Restoration of the CD4+CD25bright Regulatory Cell Subset in Patients with Crohn’s Disease Following Autologous Non-Myeloablative Stem Cell Transplantation (NST).
Authors: Verda Larissa , Oyama Yu , Craig Robert , Statkute Laisvyde , Krosnjar Nela , Quigley Kathleen , Burt Richard K. 
Published: October 2019

Abstract
Regulatory CD4+CD25 bright cells are a distinct population of T cells which modulate Th1 and Th2 mediated immune responses and have a preventive role in the development of autoimmune diseases including Crohn’s disease. Using flow cytometry we analyzed the percentage and absolute number of CD4+CD25bright cells in the peripheral blood of Crohn’s disease (CD) patients before and following non-myeloablative stem cell transplantation. All patients (N=14) survived the NST procedure and showed marked clinical and histological improvements in disease severity. Prior to NST, both the frequency and absolute number of CD4+CD25 bright cells in CD patients were reduced compared to healthy volunteers (p<0.01). Following NST, a robust increase in the absolute number of CD4+CD25+ cells was observed: pre-NST 10.3 ± 3.5 (N=14) vs. 6 mo post-NST 28.1 ± 13.4 (N=13) vs. 1 yr post-NST 24.7 ± 12.3 (N=12) versus 2 yr post-NST 20.2 ± 10.2 (N=9); p < 0.01. Two patients who relapsed after NST had less prominent increases in CD4+CD25 bright cell subset. When these patients were excluded from analysis, the difference between pre and post NST measurements increased (p < 0.0001). We conclude that autologous non-myeloablative stem cell transplantation induces the recovery of CD4+CD25 bright regulatory T cells which may contribute to the restoration of immunological homeostasis and remission of disease.
Absolute number of CD4+CD25+ cells in PB of CD patients pre and post NST Absolute number of CD4+CD25+ cells in PB of CD patients pre and post NST