bluuebunny/crossref_metadata_2025_split huggingface dataset:
Early Diagnosis and Treatment of the Patients with Acquired Hemophagocytic Lymphohistiocytosis
Authors: Wang Zhao , Wang Yini , Feng Cuicui , Tian Liping 
Published: October 2019

Abstract
Acquired hemophagocytic lymphohistiocytosis (HLH) is a life threatening condition characterized by uncontroling hyperinflammation on the basis of various infection, tumor and inherited immune deficiency. Awareness of the clinical symptoms and of the diagnostic criteria of HLH is crucial in order not to overlook HLH and to start life-saving therapy in time. In this study, we reviewed 57 suspected HLH patients from March 2006 to June 2008. 25 healthy subjects were enrolled in the study as control. NK cell activity in peripheral blood was tested by a released LDH assay. Meanwhile, solution interleukin-2 receptor (sCD25) was examined with ELISA double antibody sandwich assay. The level of glycosylated ferritin was also detected and the ratio of glycosylated ferritin to ferritin was determined. 41 out of 57 patients were definitely diagnosed according to HLH-2004 diagnostic criteria in this study and 16 patients were excluded. We found that the level of NK cell activity and the ratio of glycosylated ferritin in the all 41 final diagnozed HLH patients were significantly lower than those in the 16 excluded patients and 25 healthy control subjects (p<0.01). Meanwhile, the level of sCD25 in peripheral blood was much higher in all the 41 HLH patients than that in the excluded and healthy people (p<0.05). We compared the coincidence of each diagnostic index in the 41 HLH patients before and after final diagnosis. It was found that 100% patients had abnormal expression on NK cell activity, sCD25 and glycosylated ferritin in the early disease. The three diagnostic indexes were more sensitive and specific than other indexes, such as fever, hepatosplenomegaly, cytopenia, hyper-triglyceridemia, hypo-fibrinogenemia. 41 diagnosed patients received the regimen containing methylprednisolone and immunoglobulin, with or without fludarabine, 26 out of 41 were markedly improved after treatment, 10 out of 41 were exacerbated, and other 5 patients gave up treatment. It is concluded that detection of NK cell activity, sCD25 and glycosylated ferritin may play a very important role in the early diagnosis of HLH. Our data also suggest that fludarabine combined with methylprednisolone and immunoglobulin (FDIg) may provide a new viewpoint for HLH therapy.
PU.1 Plays a Critical Role in Maintenance of Mature Dendritic Cell Pool
Authors: Iino Tadafumi , Chong Yong , Mizuno Shin-ichi , Ito Kyoko , Tenen Daniel G. , Reizis Boris , Akashi Koichi 
Published: October 2019

Abstract
PU.1, a hematopoietic transcription factor, is absolutely required for development of myelo-lymphoid cells from hematopoietic stem cells (HSC). PU.1-deficient mice fail to develop common myeloid progenitors (CMPs) or common lymphoid progenitors (CLPs), resulting in complete loss of dendritic cells (DC) in addition to mature myeloid and lymphoid cells. In this study, by disrupting PU.1 specifically at the mature DC stage, we here show that PU.1 is necessary for maintenance of mature DC pool. By crossing PU.1 floxed/floxed mice with a mouse line harboring the Cre transgene driven by the CD11c-BAC, we disrupted PU.1 at the CD11c+ DC stage. In these mice, development of DC precursors such as Lin−c-KitloFLT3+MCSFR+ DC progenitors, FLT3+ CLP and FLT3+CMP were not affected. The number of CD11c+B220− DCs, however, significantly reduced in all lymphoid tissues including the thymus, the spleen, the lymph node and the skin, down to <30%, <10%, <10% and <5% of DC numbers in control mice, respectively. In contrast, mice possessed normal numbers of granulocytes/monocytes, B cells, and naïve, effector or regulatory T cells. These mice have not developed any significant hematological or immune disorders at least until 6 months after birth. These results clearly show that PU.1 is required not only for DC development but also for maintenance of the peripheral DC pool. We are currently trying to elucidate the underlying mechanism for PU.1 to maintain mature DC numbers in peripheral organs.
Spatial Analysis of Hematopoietic Stem and Progenitor Cells in the Bone Marrow
Authors: Nombela-Arrieta Cesar , Harley Brendan , Levantini Elena , Mahoney John E , Pivarnik Gregory , Protopopov Alexei , Tenen Daniel G. , Silberstein Leslie E. 
Published: October 2019

Abstract
Sustained production of all mature blood cell types relies on the continuous proliferation and differentiation of a rare population of self-renewing, multipotent hematopoietic stem cells (HSCs). HSC maintenance and lineage differentiation are strictly regulated by distinct microenvironments, termed niches, defined by cellular components, soluble regulators, and by the extracellular matrix. Definitive identification of the location as well as cellular and extracellular characteristics of HSC niches in the bone marrow (BM) has not been completed due to limitations of conventional imaging techniques. We have employed a novel imaging technology, Laser Scanning Cytometry (LSC) to define the localization of hematopoietic stem and progenitor cells (HSPCs) within different regions of the BM. LSC allows imaging and objective quantitative analysis of the anatomical position(s), number, and frequency of specific cell populations within the native tissue microenvironment. Analysis of whole femoral longitudinal sections of Bmi-GFP mice, in which GFP is expressed at its highest levels in HSPCs, revealed that within the bone diaphysis, HSPCs (Bmi-GFPhi c-kit+) cells were highly enriched in endosteal regions (within 100nm away from inner bone surface) compared to the central medullary region. Importantly, our data show that HSPCs are found at highest frequencies in the metaphysis of long bones, suggesting that these areas, which display characteristic morphological features, are functionally distinct from the diaphyseal region and a preferential location for HSPC-specific niches. We are currently employing LSC to identify HSPC niche cellular constituents by quantifying the relative frequency at which these cells are found in association with previously proposed niche-components such as osteoblasts, BM endothelial sinusoidal cells and CXCL12-abundant reticular cells. A detailed understanding of niche-derived signals regulating unique properties of HSCs will certainly prove relevant in human HSPC transplantation and cell therapy.
High-Dose Chemotherapy and Autologous Stem-Cell Transplantation for Primary CNS Lymphoma: Updated Results from a Pilot and Phase II Study
Authors: Illerhaus Gerald , Müller Fabian , Feuerhake Friedrich , Finke J.ürgen 
Published: October 2019

Abstract
Introduction: High-dose chemotherapy (HDT) and autologous stem-cell transplantation (ASCT) demonstrated high efficacy in the treatment of newly-diagnosed primary CNS lymphoma (PCNSL) in younger patients (pts.). A 5-year overall survival probability (OS) of 69% could be demonstrated in 30 pts within a phase-II trial on HDT and ASCT with consolidating whole-brain-irradiation (WBRT) (Illerhaus et al. JCO 2006). A subsequent pilot trial on HDT and ASCT without WBRT showed a 5-year OS of 77% (Illerhaus et al. Haematologica 2008). Here we give an update of our two different treatment regimens and future perspectives.
Patients and Methods: Thirty pts. ≤65 years were treated within the phase II trial, chemotherapy (CHT) consisted of 3 cycles of high-dose methotrexate (MTX, 8 g/m2), 1 cycle of AraC (2× 3 g/m2) plus thiotepa (TT, 40 mg/m2) followed by rG-CSF and stem-cell-mobilization. Conditioning regimen included BCNU (400 mg/m2) and TT (2×5 mg/kgBW) followed by ASCT. Hyperfractionated WBRT (45 Gy, 2×1Gy/d) was administered as consolidation. In our subsequent pilot trial 13 pts. (age 38–67 years) were treated without consolidating WBRT; CHT was intensified with 4 cycles MTX 8g/m2, 2 cycles AraC (2× 3 g/m2) and TT (40 mg/m2). Dose escalated HDT included BCNU (400 mg/m2) and TT (4×5 mg/kgBW) followed by ASCT. WBRT was reserved for pts. not responding to CHT.
Results: Median follow-up of the 30 pts. treated within our phase II trial was extended to 95 months (mo), the updated 5-year OS of all pts. is 66.6% and 82,3% of the subgroup of pts. who underwent HDT and ASCT (n=23), respectively. Three additional deaths occurred due to relapse (n=2) after 45 and 71 mo and due to comorbidity (n=1) after 103 mo. Five of 30 pts. developed severe leukoencephalopathy during follow-up. With a median follow-up of 35 mo in the 13 pts. treated within the pilot-phase without consolidating WBRT 3-year OS of all pts. is 77%. No further relapse or non-relapse mortality occurred in this pilot-group during. Most recent follow-up data will be presented in detail.
Conclusion: Sequential systemic application of high-dose cytostatic agents followed by HDT+ASCT is highly effective as initial therapy for pts. with PCNSL. The restriction of WBRT to refractory disease shows similar OS rates and a decrease in neurotoxicity. In an ongoing multicenter phase-II trial immunotherapy with rituximab is combined with HDT and ASCT to further increase remission rates. A future randomized trial should be focused on the efficacy of consolidation with HDT supported by ASCT.
Gem-(R)CHOP Versus (R)CHOP: A Randomized Phase II Study of Gemcitabine Combined with (R)CHOP in Untreated Aggressive Non- Hodgkin’s Lymphoma - EORTC Lymphoma Group Protocol 20021
Authors: Aurer Igor , Eghbali Houchingue , Raemaekers John M. , Khaled Hussein M , Van Glabbeke Martine , Baila Liliana , van der Maazen Richard 
Published: October 2019

Abstract
Despite recent advances many patients with aggressive NHL succumb to their disease and improvements in front-line chemotherapy are still needed. Gemcitabine is active in lymphoma. We embarked on a randomized phase II trial to determine the efficacy, feasibility and toxicity of the addition of gemcitabine to standard front-line chemotherapy. The trial consisted of two parts. The first part was designed to determine the maximal tolerated dose of gemcitabine and the second to determine the response rate, feasibility and toxicity of the treatment. Patients, 18–70 years old, with stage II–IV previously untreated B-large cell, follicular grade 3, anaplastic large cell or peripheral T cell lymphoma were eligible. They were randomized to receive either 8 cycles of standard CHOP given every 3 weeks or the same treatment combined with gemcitabine (Gem-CHOP). In the second part of the trial patients with B-NHL also received 8 cycles of rituximab (R). The starting dose of gemcitabine was 800 mg/m2 on days 1 and 8. Dose escalation to 1000 and 1250 mg/m2 and reduction to 800 mg/m2 on day 1 only were foreseen. Twenty-five patients, 7 with T and 18 with B-NHL, were enrolled in the trial; 15 in the dose-finding part and 10 in the second part before early closure due to low accrual. Twelve received Gem-(R) CHOP and 13 (R)CHOP. Maximal tolerated dose of gemcitabine was 800 mg/m2 given on days 1 and 8; dose limiting toxicity was hematologic. Five patients (42%) treated with Gem-(R)CHOP achieved CR in comparison to 10 (77%) treated with (R)CHOP. Three out of 4 patients with T-NHL responded to Gem-CHOP and 2 out of 3 to CHOP. Median time to treatment failure was 1.49 years for the Gem-(R)CHOP arm and 3.13 years for the (R)CHOP arm. Four patients receiving Gem-(R)CHOP had serious pulmonary toxicity, as compared to none receiving (R)CHOP. One patient died of severe pneumonitis. The apparently immunologically mediated pulmonary toxicity prompted us to add in the second part of the trial a three-day course of prednisone starting on day 8, together with the second gemcitabine dose. After this change, no additional cases of pulmonary toxicity were seen. The combination of gemcitabine with standard (R)CHOP results in significant but acceptable hematologic toxicity. Maximal tolerated dose of gemcitabine is 800 mg/m2 given on days 1 and 8 every 3 weeks. In this small group of patients, addition of gemcitabine did not seem to improve outcomes. The use of gemcitabine in previously untreated patients with aggressive NHL results occasionally in severe, potentially fatal, pulmonary toxicity. Therefore, further exploration of this combination does not seem to be warranted.
Combination Therapy with Histone Deacetylase Inhibitor Vorinostat Plus Aurora Kinase Inhibitor MK-0457 Leads to Enhanced Lymphoma Cell Killing with Stabilization of p53 and Repression of C-Myc, hTERT, and Myc-Responsive miRNAs
Authors: Kretzner Leo , Scuto Anna , Jove Richard , Forman Stephen J. , Yen Yun , Kirschbaum Mark 
Published: October 2019

Abstract
Background: Patients with relapsed or refractory Hodgkin (HL) and Non Hodgkin Lymphoma (NHL) have few therapeutic options after salvage therapy and transplant, and new agents are thus needed. MK-0457 is a novel aurora kinase inhibitor (AKI) which leads to polyploidy and G2 arrest by disregulating mitosis. We demonstrate that the addition of vorinostat, a broad spectrum HDACi, leads to the reactivation of proapoptotic and DNA damage response elements leading to enhanced lymphoma cell death. A panel of HL and NHL cell lines was studied with either drug or the combination using MTS, Annexin V, and flow cytometry studies.
Results: Vorinostat alone at a dose range of 0.5 – 3 mM reduces cell growth by at least 50% in all lines tested. For example, 1.5 mM vorinostat results in averages of 50% growth reduction and 30% apoptotic cells at 48 hours in the L540 HL cell line. MK-0457 alone at 100 nM – 500 nM results in up to a 50% reduction in cell growth and 20% apoptosis in the tested cell lines. However, the combination of 1.5 mM vorinostat and 100 nM MK-0457 results in 50 – 70% apoptosis of the L540 and KM-H2 Hodgkin cell lines at 48 hours. Cell cycle analyses by FACS show an increased percentage of cells in G2/M with few cells in sub-G1, whereas in combination with vorinostat there is a significant increase in the sub G1 population. Real-time PCR analysis and immunoblotting of L540 cells treated with either single agent or in combination suggest an epigenetic as well as a protein acetylation mechanism for this activity. Prosurvival genes such as bcl-XL and hTERT are downregulated five-fold by combination drug treatment, while the proapoptotic BAK gene is upregulated 1.5 – 2-fold. The p53 tumor suppressor is stabilized both by increased acetylation in response to vorinostat as well as by reduced inhibitory phosphorylation at Ser315 by MK-0457-inhibited aurora kinase, leading to a corresponding increase in the p53 target genes p21 and Noxa. Protein and mRNA levels of c-myc are reduced. Repression of microRNAs (miRNAs) in the myc-regulated polycistronic cluster of miRNAs of chromosome 13, such as miR-17.5p, -17.3p, and 18, is demonstrated with vorinostat and TSA.
Conclusions: The HDACi vorinostat leads to both transcriptional and post-transcriptional changes which create a pro-apoptotic milieu, sensitizing the cell to centrosome-acting agents such as the AKI, MK-0457.
[We acknowledge Merck Inc for providing Vorinostat, MK-0457, and research support.]<
Cytogenetic Analysis and FISH in the Evaluation of Patients with MDS: A Retrospective Analysis of Concordance Rate, Utility and Benefit of Performing Both Studies Simultaneously
Authors: le Shammo Jami , Gimelfarb Alla , Hsu Wei-Tong , Paz Irene Dehghan- , Venugopal Parameswaran , Larson Melissa L. , Fung Henry C. , Farhat Mohamed , Rizman Amy , Loew Jerome , Gregory Stephanie A. 
Published: October 2019

Abstract
Background: The Myelodysplastic syndromes (MDS) are hematopoietic stem cell disorders, characterized by cytopenia, and risk of progression to acute leukemia. Chromosomal abnormalities are detected in 40–60%of patients with MDS. Cytogenetic findings are critical for diagnosis, prognosis and monitoring therapy. FISH has been reported to detect occult clonal abnormalities in 15–7.8% of karotypically normal patients in some studies, but found to be of limited value in others. We sought to evaluate whether FISH can be a valuable diagnostic/prognostic adjunct to conventional cytogenetic (CC) analysis.
Methods: Since the implementation of FISH panel at our institution, we have identified 62 MDS patients who had both CC and FISH panel tests performed simultaneously. Our FISH panel was designed specifically to detect abnormalities in chromosomes 5, 7, 8, 11 and 20.
Results: In these patients, concordance between the 2 methods was noted in 43 patients (69.3%). Clonal chromosomal abnormalities were detected by CC in 36 patients (58 %). FISH defined additional clones in 7/26 patients with normal and 3/36 patients with an abnormal karyotype increasing the detection rate by 16%. Importantly, 9/36 patients with abnormal CC had clonal abnormalities in chromosome regions not analyzed, thus not detected, by our FISH panel.
To further evaluate the prognostic significance of the additional clones detected by FISH, we calculated and compared the IPSS risk category for the 10 patients identified with discordant results utilizing data derived from CC and FISH. 3 patients had no change in their IPSS scores, 7 patients had an upgrade in their IPSS scores as follows: from low to Int-1: 4, Int-1 to Int-2: 2, and Int-2 to High: 1. If one were to apply NCCN guidelines with regard to treatment options according to IPSS risk categories, FISH upgraded scores could potentially result in change of therapy in 2/10 patients with discrepancy, and in 2/62 (3%) patients altogether.
Conclusion: Our results suggest that FISH has the potential of increasing the rate of detecting chromosomal abnormalities in complete CC studies that have failed to reveal clonal abnormalities. However, the practical impact of finding such clones seems to be very low resulting in potential change of clinical management in only 3% of patients. Furthermore, in cases with identified cytogenetic abnormalities, FISH was of limited benefit. Our series is small and retrospective in nature; prospective studies addressing the utility, benefit, and cost justification of this method are needed.
The Immunotoxin HM1.24-ETA′ Is a Potent Inducer of Apoptosis in Malignant Plasma Cells
Authors: Staudinger Matthias , Muskulus Anja , Burger Renate , Guenther Andreas , Repp Roland , Gramatzki Martin , Peipp Matthias 
Published: October 2019

Abstract
Despite new treatment modalities, the clinical outcome of at least a subgroup of patients with multiple myeloma (MM) still needs improvement. Antibody-based targeted therapies are increasingly used for tumor therapy, and may represent interesting options for MM patients. HM1.24 is a surface molecule that is over expressed on malignant plasma cells and efficiently internalized from the cell surface. It may represent a promising target for the development of myeloma-directed immunoconstructs. Here, the development and characterization of a novel single-chain immunotoxin, HM1.24-ETA′, is described. HM1.24-ETA′ was generated by genetic fusion of an HM1.24-specific single-chain Fv (scFv) antibody and a truncated variant of Pseudomonas aeruginosa exotoxin A (ETA′). The immunotoxin was expressed in E. coli and purified to homogeneity by affinity chromatography. Specific binding to HM1.24 was demonstrated by immunofluorescence staining and flow cytometry using antigen positive and negative cells. HM1.24-ETA′ efficiently inhibited growth of IL-6 dependent and IL-6 independent myeloma cell lines (INA-6, RPMI8226, U266). Half maximal growth inhibition was observed at low nanomolar concentrations. Further analyses demonstrated that target cell killing occurred via induction of apoptosis, as evidenced by Annexin V/propidium iodide staining and detection of PARP cleavage. The cytotoxic effect was completely blocked by adding excess of unconjugated parental antibody, demonstrating that the effect was antigen-specific and not mediated by non-specific uptake of the immunotoxin. Importantly, HM1.24-ETA′ efficiently triggered apoptosis (>80% Annexin V positive cells) of freshly isolated plasma cell leukemia cells within 48h. In conclusion, HM1.24-ETA′ efficiently triggered apoptosis of multiple myeloma cell lines as well as freshly isolated tumor cells. These results indicate that HM1.24 may represent a promising target structure for efficient antigen-specific delivery of cytotoxic compounds to plasma cell tumors.
An Animal Model to Study Bisphosphonates Associated Osteonecrosis of the Jaw
Authors: Pozzato Gabriele , Chiandussi Silvia , Zacchigna Serena , Dore Franca , Moimas Silvia , Zanconati Fabrizio , Cavalli Fabio , Mecchia Arturo , Faraon Michela , Giacca Mauro , Di Lenarda Roberto , Giacca Mauro , Biasotto Matteo 
Published: October 2019

Abstract
The pathogenesis of osteonecrosis of the jaws (ONJ) in patients treated with intravenous bisphosphonates is still not completely understood, and likely involves both reduction in blood supply and the activity of oral bacteria. We present an animal model of ONJ, which will significant contribute to the understanding of the aetiopathogenesis of this condition. Five skeletally-mature female Wistar rats (weight 450 g) were given intravenous zoledronic acid 0.04 mg in saline solution (0.2 mg/ml) once a week for 5 weeks. After 2 weeks, the animals underwent the extraction of the upper right first molar, followed by the creation of a 4 mm-diameter bone defect on the same site under general anaesthesia. After 7 weeks from the extraction, the animals were clinically examined and a static bone-scintigraphy using 99mTc-MDP methylene diphosphonate was performed with a gamma camera equipped with pinhole collimator (Siemens-Ecam). After an additional week, the rats were sacrificed and a Computed Tomography (CT) was carried out. Samples obtained from the bone defect were decalcified and prepared for histological assessment. Five rats, not treated with zoledronic acid and exposed to the same surgical treatment, were used as controls. Histological and imaging features were assessed blindly. At 7 weeks after the extraction, all the rats treated with zoledronic acid showed impair healing, expansion of the defect and bone exposure. These features were confirmed by the scintigraphy, which showed abnormal localized activity in comparison with the surrounding tissues. On clinical examination, the rats of the control group demonstrated epithelialization of the bone defect and a normal uptake of the contrast medium during the scan. The CT scan disclosed irregularity of the cortical margin and destruction of the cortical bone, which were not evident in the control group. The histology showed sheets of necrotic bone, with loss of osteocytes from their lacunae and peripheral resorption. No inflammatory infiltrate was observed. The control group instead demonstrated normal bone healing. All animals were treated according with the guiding principles for experimental procedures found in the Declaration of Helsinki of the World Medical Association. Based on this study, the rat treated with zoledronic acid can be considered a novel, reliable and reproducible model to better understand the pathophysiology of ONJ and to develop a therapeutic approach.