bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immune Hemolysis and Aseptic Meningitis after Intravenous Immunoglobulin
Authors: Arumugam Raajasekar Arun Kumar , Nannapaneni Srikant , Solomon William B. 
Published: October 2019

Abstract
INTRODUCTION
Human intravenous immunoglobulin (IVIG) products are generally considered safe and are used for the treatment of a variety of autoimmune and pro-inflammatory states. However, controlled trials involving IVIG have been of small size with limited power and described only the most common adverse effects [1]. IVIG related hemolysis is a rare adverse effect and is usually self-limited, rarely requiring blood transfusions. A few case reports have also described aseptic meningitis after IVIG use. We describe a patient who developed severe hemolytic anemia and aseptic meningitis after IVIG use.
CASE PRESENTATION
A 54 year old woman was admitted with severe head ache, dark colored urine and shortness of breath twenty four hours after receiving IVIG. She was diagnosed with chronic inflammatory demyelinating polyneuropathy (CIDP) a month ago and was started on IVIG. Physical exam was unremarkable. Laboratory tests showed severe anemia with a hemoglobin of 6.1 g/dl (baseline 13 g/dl), indirect hyper-bilirubinemia, low haptoglobin, elevated LDH and hemoglobinuria confirming hemolysis. Hematological testing revealed a positive direct antiglobulin test (DAT), DAT IgG, DAT CD3 and eluate testing were also positive. A lumbar puncture showed neutrophil pleocytosis but no bacteria. A diagnosis of IVIG induced hemolysis and aseptic meningitis was made. O negative packed red blood cells were transfused for severe symptomatic anemia and non-steroidal anti-inflammatory drugs were given for headache. Steroids were deferred since the source of antibodies causing hemolysis was extrinsic. She improved clinically and blood counts returned to baseline in three weeks.
DISCUSSION
Pooled IVIG is extracted from at least 1000 individuals and contains highly purified polyvalent IgG [2]. Hemolysis after IVIG is thought to be secondary to donor antibodies against host red blood cell antigens and resolves once IVIG is stopped. Risk factors for hemolysis include non-O blood types and the use of high dose IVIG. The patient described was of AB Rh positive blood type but did not receive high dose IVIG. Aseptic meningitis is another rare adverse effect that has been postulated to be related to antibodies in the IVIG that mimic antineutophil cytoplasmic antibodies (ANCA) which activate neutrophils causing neutrophilic pleocytosis [3]. One must be aware of these two rare complications of a commonly used drug.
REFERENCES
[1] Risks associated with the use of intravenous immunoglobulin. Pierce LR et al. Transfus Med Rev. 2003 Oct;17(4):241-51.
[2] Appropriate uses of human immunoglobulin in clinical practice: memorandum from an IUIS/WHO meeting. Bull World Health Organ. 1982;60(1):43.
[3] Intravenous immunoglobulins contain naturally occurring antibodies that mimic antineutrophil cytoplasmic antibodies and activate neutrophils in a TNF alpha-dependent and Fc-receptor-independent way. Jarius et al. Blood. 2007;109(10):4376.

Disclosures
No relevant conflicts of interest to declare.
Presence of Primary Cilia in Human Hematopoietic System
Authors: Singh Mohan , Chaudhry Parvesh , Ramsingh Giridharan , Merchant Akil 
Published: October 2019

Abstract
Primary cilia are non-motile microtubule-based organelles that are present on the cellular membrane of all eukaryotic cells. The presence of cilia is required for the response to developmental signaling pathways such as Hedgehog (Hh) and Wnt/β-catenin. Loss of primary cilia or their function has been linked to a number of human genetic diseases, called ciliopathies (Dorn et al., Dev Cell, 2012). Recent studies suggest that abnormal hedgehog Hh pathway activity due to ciliary dysfunction could lead to various cancers such as glioblastoma, medulloblastoma, basal cell carcinoma, breast, prostate and ovarian cancer (Menzl et al., Cilia, 2014, Moser et al., BMC cancer 2009). Although the Hh pathway has emerged as an exciting new therapeutic target in leukemia and myeloproliferative diseases, to date, there have been no reports describing the presence of primary cilia in human blood or leukemia cells. In the present study, we have successfully identified primary cilia in hematopoietic system (Fig. 1). Primary cilia are well characterized in hedgehog responsive NIH3T3 cells; therefore, these cells were included in the study as a positive control for cilia staining. Localization of acetylated tubulin (which stains for the ciliary body) and gamma tubulin (which stains the basal body) was used to confirm the presence of primary cilia in normal human peripheral blood mononuclear cells (PBMCs) and normal bone marrow (BM) cells. Furthermore, we have also identified primary cilia in acute myeloid leukemia cell lines (KG1, KG1a, K562). Additionally, hedgehog pathway components such as, Smoothened and Gli3 were localized in primary cilia of acute myeloid leukemia (AML) cell lines. Interestingly, the frequency of primary cilia in AML cells (10-36%) was found to be lower than normal PBMCs and BM (97-99%) (Fig. 1). Moreover, cilia present on leukemic cells were often shorter and dysmorphic, with less well defined basal bodies. KG1 cells, which have a higher number of cilia, retain sensitivity to Hh pathway inhibitors, but K562 and KG1a, which have high levels of Hh activity but very low numbers of cilia, are resistant. Loss of cilia is, therefore, associated with activation of Hh signaling that is resistant to SMO antagonists, and could have important implications for the development of this class of drugs. In conclusion, we present the first evidence showing the presence of primary cilia in hematologic system and implicate the loss of primary cilia in development of hematological malignancies through deregulated Hh signaling.

Disclosures
No relevant conflicts of interest to declare.
Abnormal Levels of PKM2 in Mdcs from Patients with Severe Aplastic
Authors: Shao Zonghong , Zheng Mengying 
Published: October 2019

Abstract
Severe aplastic anemia (SAA) is a hematologic disease characterized by pancytopenia with severe bone marrow failure. Our previous studies have demonstrated that activated myeloid dendritic cells (mDCs) increased in the bone marrow of SAA patients, which might promote Th0 cells to polarize to Th1 cells and cause the subsequent over-function of T lymphocytes and hematopoiesis failure in SAA. The other notable finding in our study is that abnormal expression of pyruvate kinase M2 (PKM2) in mDCs from SAA patients may be one of the reason for mDC hyperfunction. Human pyruvate kinase M2, which catalyzes the last but rate-limiting step of glycolysis, is exclusively expressed in actively proliferated cells ,especially in embryonic and adult dividing/tumor cells. It remains unclear how PKM2 may change in patients with SAA. This study aims at exploring the role of PKM2 in SAA. In this study, the quantitative changes of mDCs in peripheral blood of 15 SAA patients before immunosuppressive therapy (IST) °¢ 13 SAA patients after IST and 26 normal controls were determined by flow cytometry. Results showed that the levels of PKM2 in mDCs were significantly increased in SAA patients(59.1±15.8)%. After IST, the levels of PKM2 in mDCs decreased(42.6±22.1)% (P<0.05).In normal controls group , the level of PKM2 in mDCs was (32.7±20.2)%. These changes reveal that dysregulation of PKM2 expression and activation in mDCs from SAA patients may contribute to hyperfunction of mDCs, thus leading to the imbalance of downstream Th1/Th2 subsets which followed by hematopoiesis failure in SAA.
Figure 1. Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Tumor Microenvironment Regulates Survival of Mantle Cell Lymphoma Cells through Various Signaling Pathways
Authors: Sehgal Lalit , Tamer Khashab , Wang Xin , Mathur Rohit , Berkova Zuzana , Romaguera Jorge E. , Rodriguez Alma , Neelapu Sattva S. , Samaniego Felipe 
Published: October 2019

Abstract
Introduction
An aggressive, incurable form of non-Hodgkin's lymphoma (NHL) is classifed as Mantle cell lymphoma (MCL). The health complications associated with advanced age of restrict treatment with intense chemotherapy. Overexpression of cyclin-D1 due to Translocation t(11;14) , is the hallmark of MCL. More detailed insight into MCL pathogenesis has been delayed until the recent development of a tissue culture system, using human mesenchymal stromal cells (hMSC), suitable for propagating primary MCL cells. We hypothesized that tumor-initiating cells are responsible for MCL relapse and chemoresistance and thus, identification of signals responsible for survival and maintenance of MCL-initiating cells (MCL-ICs) is essential for design of curative treatment strategies.
Methods
Isolates of primary MCL cells (n=24) were co-cultured with human mesenchymal stem cells (hMSCs) and the content of MCL-ICs was analyzed by flow-cytometry based on marker expression profile; CD34-CD3-CD45+CD19-. Cytokine array was used to identify the soluble factors enriched in the co-cultures and the expression of these factors was confirmed by RT-PCR analysis. The signaling pathways employed by the newly-identified factors were blocked in 3 MCL cell lines (JVM2, Mino, Z138) to confirm their essential role in survival of MCL cells and, more importantly, for MCL-ICs.
Results
Co-cultures of primary MCL isolates with hMSCs supported the growth of MCL cells for over 4 weeks with continued presence of MCL-ICs (CD34-CD3-CD45+CD19-) representing about 1% of MCL cells. We found that IL-6 produced by hMSCs triggered an FGF/FGFR autocrine loop in MCL-ICs. The extent of FGFR expression correlated tightly with expression of SOX11, a pathology related negative prognostic marker in MCL. MCL cell survival and growth was regulated via the FGFR/mir101/ EZH2/ NF-κB/XIAP axis. Blocking of this signaling pathway with FGFR1 inhibitors consistently induced early reduction in XIAP levels and subsequently MCL cell death.
Conclusion
We established that propagation of primary MCL in co-cultures with hMSCs depends on an FGF/FGFR autocrine loop that enhances XIAP protein expression and thus, supports survival of MCL cells. We identified the factors essential for survival of MCL and MCL-ICs that present new targets for improved MCL treatment strategies.

Disclosures
Rodriguez: Orthobiotech: Research Funding.
Improved Molecular Diagnosis of Aplastic Anemia/Bone Marrow Failure Syndromes Using a Novel Comprehensive Next Generation Sequencing Gene Panel
Authors: Dasouki Majed , Ahmed Syed Osman , Alahmari Ali , Jabr Amal , Alawwami Moheeb Ali , Chaudhri Naeem A. , Fahad AlMohareb , Mohamed Said Y , Rasheed Walid , Alzahrani Hazza A , Aljurf Mahmoud 
Published: October 2019

Abstract
Introduction: Various forms of aplastic anemia (AA)/bone marrow failure syndromes (IBMFs) show significant clinical and molecular heterogeneity with significant clinical overlap and are often diagnosed based on established clinical and pathological criteria. While >70 genes have been identified in patients with AA/IBMFs, most cases have are labelled as idiopathic with no identifiable genetic abnormlaity found. Precise detection of genetic abnormalities in these patients may assist in more accurate molecular diagnosis in these patients, proper counseling, cancer surveillance and personalized clinical intervention.
Method: As part of the Saudi Human Genome Project, we developed a comprehensive 405 gene panel encompassing all known Mendelian hematological disorders (hemolytic anemias, aplastic anemias/bone marrow failure syndromes, coagulation disorders) using the Ion Torrent AmpliSeq technology. Patients who met the clinical diagnosis of aplastic anemia/bone marrow failure syndrome were enrolled into this study. Peripheral blood samples were subjected to this next-generation sequencing analysis.
Results: We validated the Saudi Mendeliome assay using 642 samples with known mutations across various medical specialties. We then tested 37 patients with AA/IBMFS using this Proton-Ion sequencing platform. Mutations were identified in 7/37 (19%) of patients, followed by whole exome sequencing (WES) in those patients without identifiable mutations.
Conclusion: Compared with clinical WES and/or whole genome sequencing (WGS), which are still expensive, time consuming and difficult to interpret, this novel and comprehensive targeted gene panel is more economical (< $150), faster (3-4 weeks), upgradable (by spiking in newly identified AA/IBMFs genes) and can be used to genotype patients with acquired aplastic anemia/bone marrow failure syndromes and guide their management. Second tier testing using WES/WGS is recommended for cases without identifiable mutations.

Disclosures
No relevant conflicts of interest to declare.
A Small-Molecule Suppressant of Survivin YM155 Induces Cell Death Via Proteasomal Degradation of c-Myc in Multiple Myeloma Cells
Authors: Ito Shigeki , Asahi Maki , Ishida Yoji 
Published: October 2019

Abstract
[Background] Survivin is a member of the inhibitor of apoptosis protein (IAP) family with its dual roles in mitosis and apoptosis, and emerges as an attractive target for cancer therapy. YM155, a novel molecular targeted agent, suppresses survivin, which is overexpressed in many tumor types. However, the effect of this agent on multiple myeloma (MM) cells remains unclear. [Materials & Methods] Five human MM cell lines, RPMI8226, U266, KMS20, KMS28PE, and KMS34 were used in this study. Cell proliferation and cell death were evaluated by MTT assay and by flow cytometric analysis with annexin V/PI staining. Gene and protein expressions were analyzed with quantitative PCR (qPCR) and immunoblot, respectively. For proteasome inhibitory assay, cells were treated with YM155 and/or proteasome inhibitor MG132 for 6 hours. [Results] YM155 inhibited cell proliferation of these cells in a dose- and time-dependent manner. Annexin V assay showed that YM155 induced apoptosis in these cells. To better understand these effects of YM155 on MM cells, we evaluated the intracellular signaling and apoptosis-associated protein status. Immunoblot analyses showed that YM155 reduced not only survivin but also Mcl-1 and XIAP expressions. We also observed the activation of caspase-3 and PARP in YM155-treated cells, indicating that YM155 induces caspase-dependent apoptosis. In contrast, YM155 did not affect phosphorylation status of Erk1/2 and STAT3. Interestingly, YM155 suppressed c-Myc and IRF4 protein expressions, both of which are recognized as an important transcription factor in the pathogenesis of MM. In addition, qPCR assay showed that YM155 treatment did not reduce c-Myc mRNA level. On the other hand, proteasome inhibitor prevented the suppression of c-Myc expression by YM155 treatment, indicating a proteasomal degradation of c-Myc by YM155. [Conclusion] YM155 suppresses cell proliferation and survival in MM cells in part via not only inhibiting anti-apoptotic proteins such as survivin, Mcl-1 and XIAP but also suppressing c-Myc oncoprotein expression. Further study is needed to clarify the molecular mechanism of c-Myc degradation induced by YM155. Our results may provide a new concept in c-Myc-targeting therapy for MM because c-Myc oncoprotein have been considered undruggable.

Disclosures
Ishida: Bristol-Myers Squibb: Honoraria.
Cross Talk Between Haemostasis and Oxidative Stress in Acute Leukemia
Authors: Naz Arshi , Sattar Abdul , Farzana Tasneem , Taj Mehwish , Ansari Saqib Hussain , Nadeem Mohammad , Shamsi Tahir 
Published: October 2019

Abstract
Objective: To investigate the haemostatic/oxidative changes and its association in patients of acute leukemia at the time of diagnosis.
Study Design: Cross sectional and descriptive.
Setting: Study was carried out in National Institute of Blood Disease and Bone Marrow Transplantation during 2011 to 2013.
Material and Methods: Activity of Superoxide dismutase(SOD), Glutathione Reductase(GR), Total antioxidant status(TAS), Prothrombin Time, Activated Partial Thromboplastine Time, Fibrinogen, Factor VIII and Tissue Factor were evaluated in the hemolysate, sera, citrated plasma and leukemic cells of 150 patients with acute leukemia at the time of diagnosis in 40 healthy individuals.
Results: The mean age of patients was 29.8 years.Activity of SOD, GR and TAS were significantly higher in patients of acute leukemia as compared to control (<0.01). Significant haemostatic changes were identified in patients of acute leukemia. Strong association was found between oxidative stress/total antioxidant status markers and haemostatic parameters which are in favor of cross talk between haemostasis and oxidative stress in acute leukemia.

Disclosures
No relevant conflicts of interest to declare.
A New Strategy to Target Acute Myeloid Leukemia Stem and Progenitor Cells Using Chidamide, a Histone Deacetylase Inhibitor
Authors: Xu Bing , Li Yin , Chen Kai , Zhou Yong , Xiao Yiren , Deng Manman , Jiang Zhiwu , Ye Wei , Wei Xinru , Wang Xiangmeng , Li Jie , Liang Jiabao , Zheng Zhongxin , Yao Yao , Wang Weiguang , Li Peng 
Published: October 2019

Abstract
Leukemia stem cells (LSCs) are responsible for treatment failure and relapse in acute myeloid leukemia (AML). Therefore, development of novel LSCs-targeting therapeutic strategies is of crucial clinical importance to improve the treatment outcomes of AML. Histone deacetylase (HDAC) inhibitors have emerged as a potential strategy to reverse aberrant epigenetic changes associated with cancer. HDAC inhibitors have shown potent and specific anticancer stem cell activities in preclinical studies. Chidamide, a novel benzamide-type selectively HDAC inhibitor, has been reported to induce G1 arrest and apoptosis in the relatively mature progenitor population, whereas its effect on primitive LSCs has not been clarified. In this study, we demonstrated that chidamide specifically induces apoptosis in LSC-like cells and primary AML CD34+ cells in a concentration- and time-dependent manner. Our further molecular mechanistic study uncovered that chidamide induces LSCs death by activation of reactive oxygen species (ROS). It compromises the mitochondria membrane potential, modulates anti-apoptotic and pro-apoptotic proteins in BCL2 family and activates caspase-3 leading to PARP degradation. Meanwhile, chidamide activates CD40 and modulates its downstream signaling pathways, JNK and NFκB. The results of this study suggest that chidamide may be a novel LSC-targeting agent for AML therapeutics.

Disclosures
No relevant conflicts of interest to declare.
Morphologic Evolution in Post-Transplant Lymphoproliferative Disorders (PTLD): A Clinicopathologic Case Series
Authors: Stubbins Ryan J. , Peters Anthea C , Curtis Mabilang , Owen Carolyn , Mansoor Adnan , Lai Raymond , Preiksaitis Jutta 
Published: October 2019

Abstract
Post-transplant lymphoproliferative disorders (PTLDs) are classified as early PTLDs (E-PTLD), polymorphic PTLDs (P-PTLD), monomorphic PTLDs (M-PTLD) and classical Hodgkin's Lymphoma PTLD (HL). These entities are felt to represent a disease continuum, with a precursor-product relationship, though they are morphologically and clinically distinct. However, this process remains poorly understood, and limited evidence exists to support this hypothesis. We report a series of nine cases extracted from a PTLD database, including both pediatric and adult solid organ transplant recipients, with recurrent disease episodes that evince an apparent evolution in their morphologic categorization between episodes. All patients identified were high risk for PTLD, with multiple identifiable predisposing factors. Presentations varied from isolated lymphadenopathy, to gastrointestinal involvement to pulmonary involvement. Four of the patients were deceased at the time of acquisition, though only one directly from PTLD. Eight of the nine identified patients developed E or P-PTLD lesions that preceded a subsequent M-PTLD or HL lesion at a similar tissue site. Two of the cases had metachronous P and M-PTLD lesions. The six M-PTLD subtypes were variable, including DLBCL, Burkitt, T-cell, and extramedullary plasmacytoma, in addition to the three cases of classical Hodgkin's lymphoma. These cases suggest that E and P-PTLD may act as common precursor lesions to the development of both the M-PTLD variants and HL type lesions, and that the PTLD classification schema represents different stages of a common underlying pathology.

Disclosures
Owen: Lundbeck: Honoraria; Gilead: Honoraria; Janssen: Honoraria; Roche: Honoraria.
Multiple Clonal TCR Gene Rearrangements Are Typical in Peripheral T-Cell Lymphoma Not Otherwise Specified
Authors: Sidorova Yulia V , Chernova Natalia G. , Ryzhikova Natalia V. , Smirnova Svetlana Yu. , Vinogradova Yulia E , Kovrigina Alla M. , Zvonkov Eugene E. , Sudarikov Andrey B. , Parovichnikova Elena N. , Savchenko Valeri G. 
Published: October 2019

Abstract
Introduction: Peripheral T-cell lymphoma not otherwise specified (PTCL-NOS) is a heterogeneous subgroup of PTCL with no consistent immunophenotypic, genetic and clinical features. PTCL-NOS represents less than 29% of all T-cell lymphomas in adults and often has an aggressive course with extranodal and systemic involvement. This study is focused on the application of clonality testing methods for PTCL-NOS diagnosis and monitoring.
Patients and Methods: Biopsies, blood and bone marrow samples were tested for 30 patients with PTCL-NOS: the ratio of male/female was 15/15, median age was 56 years (32-75), and 22 of 30 patients had IV stage of the disease. TCRG and TCRB gene rearrangements were evaluated for T cell clonality analysis. We used PCR-based methods (BIOMED-2 standardized clonality protocol) followed by GeneScan analysis on the ABI PRISM 3130 Genetic Analyzer (Applied Biosystems).
Results: Clonal TCR gene rearrangements were found in 29 of 30 patients (97%): TCRG in 27 of 30 (90%) and TCRB in 29 of 30 (97%). Primary biopsies or bone marrow samples showed oligoclonal pattern of rearrangements (more than 3 clonal peaks in TCRG or TCRB GeneScan profiles) in 13 of 30 patients (43%). New clonal rearrangements in affected tissues (skin, tonsil, CSF, lymph nodes and etc.) during disease progression were found in 15 of 24 (63%) patients. Various representations of clonal rearrangements in different tissues indicate the presence of several tumor clones in the majority of cases (19 of 30 patients; 63%). The presence of several clones correlated with the number of tissues examined (rs = 0.6, p <0.0005). The detection rate of additional clonal rearrangements was 90% (9 of 10 patients) when 3 and more tissues were tested. The number of clonal rearrangements was not correlated with age (p = 0.43) or the stage of disease (p = 0.29). One can speculate that the presence of multiple clones in the PTCL-NOS could reflect genetic instability of this tumor.
Conclusion: Oligoclonality, clonal evolution and the appearance of new clones in the progression of the disease are typical features of PTCL-NOS. Multiple clonal rearrangements complicate diagnostics and assessment of minimal residual disease in this disease.

Disclosures
No relevant conflicts of interest to declare.