bluuebunny/crossref_metadata_2025 huggingface dataset:
Research on the Characteristics of IGHV Gene In Chronic Lymphocytic Leukemia of China.
Authors: Qiao Chun , Miao Kourong , Li Jianyong
Published: October 2019

Abstract 4609

Objective

The usage, mutation status and prognostic impact of immunoglobulin heavy chain variable (IGHV) gene in Chinese patients with chronic lymphocytic leukemia (CLL) is unclear. We set out to define the characteristics of IGHV gene and its relevance to clinical and biological parameter in our patients.

Methods

IGHV gene mutations were detected by multiplex PCR in 202 Chinese CLL patients and the purified PCR amplification products were sequenced. IGHV somatic hypermutation status and gene usage were analyzed by IMGT/V-QUEST software. The association analysis between IGHV somatic mutation status and the clinical and biological features, including Binet staging, immunophenotype, cytogenetic aberrant, were also emphasized in this study.

Results

The results showed that 129 patients had mutated (M) IGHV, and the remaining 73 patients had unmutated (UM) IGHV according to the cutoff value of accordance rate 98%. The most frequent VH gene family was found to be VH3 (47.5%), followed by VH4 (34.7%), VH1 (11.4%), VH2 (2.5%), VH5(1.5%), VH7(1.5%) and VH6(0.9%) gene families, which was similar to other Asian populations. The overall survival (OS) time of UM IGHV group was significant shorter than M IGHV group (P=0.025). Significance was found in the expression of CD38 and ZAP-70 between patients with and without IGHV mutations (P<0.0001 and P=0.015, respectively). Binet staging was significantly different with IGHV mutation status (P<0.001). “Unmutated” sequences had significantly longer heavy chain complementarity-determining region 3 (HCDR3). Seven of these patients used VH1-69, which was similar to other Asia countries, but in striking contrast to those in Western countries, where VH1-69 was one of the most frequently used genes. FISH was performed in 117 cases, del(11q22) was considered as high risk factors, and 10(10/42, 23.8%) cases with UM IGHV gene. On the other hand, there were 7(7/75, 9.3%) cases with M IGHV gene (P=0.033). No significance was found in del(17p),del(13q),del(6q),add(12),IGH translocation between IGHV mutation and unmuatioan patients. A total of five stereotyped BCR were identified, IGHV3-21/IGHD3-9/IGHJ6, IGHV4-34/IGHD2-15/IGHJ6, IGHV1-3/IGHD6-19/IGHJ4, IGHV4-59/IGHD3-22/IGHJ6 and IGHV4-39/IGHD6-13/IGHJ5.

Conclusions

The usage of IGHV gene families indicates significant difference in Chinese CLL patients compared with Western patients, suggesting involvement of ethnic and/or environmental factors in CLL disease initiation. In the development course of CLL, BCR play an important role in the immunological recognition and selection. There are intimate relationships between mutation status of IGHV gene and prognosis. The usage of IGHV provides enlightment for the occurrence mechanism of CLL.

Disclosures:

No relevant conflicts of interest to declare.
Detection of Cytokine Protein Expression In Immune Thrombocytopenic Purpura Using Protein Arrays
Authors: Zou Shan Hua , Zhang Xuejiao , Wang Weiguang , Cheng Yunfeng
Published: October 2019

Abstract 4673

Introduction

Immune thrombocytopenic purpura (ITP) is an autoimmune disease characterized by low platelets and bleeding. The mechanism of ITP has historically been attributed to platelet autoantibody production and the resultant platelet destruction. More recent evidence suggests a multifactorial pathogenesis. Patients show impaired immune regulation manifested by increased proliferation of helper T lymphocytes and cytotoxic T lymphocytes from patients can lyse platelets in vitro. A complex picture of the immune processes involved in autoimmunity has emerged over the last decade with the identification and characterization of immunoregulatory elements (receptors, cytokines, and other signaling molecules) and cell trafficking patterns. Protein arrays are rapidly becoming established as a powerful means to detect proteins, monitor their expression levels, and investigate protein interactions and functions.

Method

During August 2009 to April 2010,17 newly diagnosed patients with ITP were enrolled. The mean platelet count at onset is 25*109/L. Seventeen health adults were as control group. We detected the changes of 507 kinds of cytokines, receptors and other signaling molecules in patients with ITP at onset, remission and relapse using the RayBio® Biotin Label-based Human Antibody Array I. The expression level differed by more than 2-fold from that observed in control group was as abnormal. We analyzed these proteins by cluster analysis.

Results

Compared with the healthy volunteers, the ITP patients at onset showed 35 elevated and 130 decreased proteins, the ITP patients at relapse showed 99 elevated and 189 decreased cytokines, the patients at remission showed 94 elevated and 144 decreased cytokines. This study shown that Th1 cytokines, JAK-STAT signaling pathway molecules, TGF-beta signaling pathway molecules, chemokine signaling pathway molecules, MAPK signaling pathway molecules and TLR signaling pathway molecules maybe closely related to the pathogenesis in ITP.

Conclusion

An understanding of the interplay of cytokines, receptors and signaling molecules in the breakdown of self-tolerance has brought to light unrecognized mechanisms of the autoimmune destruction of platelets in ITP and potential targets for future therapeutic advances. These identified proteins may constitute novel biomarkers of ITP and might hold great potential as surrogate markers for monitoring the patients with ITP.

Disclosures:

No relevant conflicts of interest to declare.
The Importance of a Bone Marrow Biopsy to Distinguish Primary Immune Thrombocytopenia From Indolent Haematological Malignancies
Authors: Gudbrandsdottir Sif , Hasselbalch Hans Carl , Frederiksen Henrik
Published: October 2019

Abstract 4676

Background

Primary immune thrombocytopenia (ITP) is characterised by an immune-mediated destruction of platelets and impaired platelet production [1, 2].

The diagnosis is one of exclusion, and any disorder presenting with isolated thrombocytopenia can mimic ITP [3].

Aims

To identify and describe patients initially diagnosed with ITP and included in an on-going clinical trial, when further diagnostics revealed other underlying disorders as the aetiology to the low platelet counts.

Methods

Patients were identified from the on-going clinical trial ‘A randomised fase III study of the efficacy of rituximab in combination with dexamethasone vs dexamethasone in newly diagnosed patients with ITP’. All patients had given their written informed consent. The diagnosis of ITP was defined as isolated thrombocytopenia, normal bone marrow biopsy, no splenomegaly assessed by ultrasound, normal TSH-levels and exclusion of secondary immune-, viral- or drug-induced thrombocytopenia.

Patients who were enrolled in the study but subsequently diagnosed with other underlying disorders and excluded from the protocol are described in the following.

Results

116 patients had been enrolled in the clinical study at the time of these analyses. 9 patients were subsequently diagnosed with other disorders and excluded from the study.

The median time from inclusion in the ITP-protocol to diagnosis of the underlying disorder was 11 days (range 3 – 486 days).

4 patients were diagnosed with other haematological disorders (debut symptoms, see figure 1).

4 patients with CMV presented median platelet count of 6 ×109/L (range 4–12 ×109/L), petechiae (4 patients) and mucosal bleeding (3 patients). 1 patient had flu-like symptoms and elevated leucocyte count. CMV was diagnosed by positive IgM titer after a median of 17,5 days (range 7 –29).

1 patient was diagnosed with antiphospholipid syndrome 17 days after inclusion. Conclusion

The diagnosis of ITP is one of exclusion, and it is essential that patients presenting with isolated thrombocytopenia be thoroughly tested for underlying disorders. The present study has shown that a bone marrow biopsy may be a prerequisite and the only diagnostic tool to distinguish ITP from MDS or other low-grade malignancies. A bone marrow biopsy is recommended in all elderly patients in whom indolent haematological malignancies initially may mimick ITP.

Disclosures:

Gudbrandsdottir: GlaxoSmithKline: Research Funding; Amgen: Research Funding. Frederiksen:GlaxoSmithKline: Membership on an entity's Board of Directors or advisory committees.
Successful Treatment of An Elderly Patient with Refractory Chronic Immune Thrombocytopenic Purpura with Combination Therapy of Rituximab and Corticosteroids: A Case Report
Authors: Ling Yun , Cao Xiangshan , Qian Xinyu
Published: October 2019

Abstract 4686

Immune thrombocytopenic purpura (ITP) is an autoimmune disorder. A corticosteroid, usually prednisone is often the first line treatment for ITP. However, the problem is that about 70 percent of adult patients experience a relapse after discontinuation of corticosteroids. And approximately 20–30% of patients with chronic ITP do not respond to corticosteroids therapy. Rituximab has been proven effective in refractory chronic ITP, but the timing of response is slower than expected, at least three months might be necessary to observe an effect. And the response duration to rituximab remains relatively short in some patients. So, sometimes it is necessary to use combination therapy including rituximab, according to different patient conditions. Here, we report an 82-year-old man with chronic ITP who had thrombocytopenia (platelet count <10 × 109/L) for more than 6 months and relapsed on a prednisone taper. He presented sustaining blood-tinged sputum, bleeding in skin and steroid-induced diabetes, the result of short-term prednisone. He didn't want splenectomy and other immune suppressive drugs. His blood glucose got control after insulin therapy. We gave the patient intravenous infusions of rituximab 375 mg/m2 weekly for 4 weeks combined with dexamethasone (10 mg intravenously weekly for 4 weeks). After first dose of dexamethasone followed by rituximab, within 24 hours his platelet count had increased to 65 × 109/L and bleeding symptoms were significantly improved. During the next 3-week period of treatment, his platelet counts fluctuated between 30 × 109/L and 60 × 109/L. And then the platelet count dropped back to a minimum of 19 × 109/L. Consider the slow responses to rituximab and prevention of bleeding, we still gave the patient maintenance therapy with 15mg prednisone daily and he had been no bleeding. Two months after starting rituximab and dexamethasone, his platelet counts began to gradually recover to normal. Although we need to further observe the patients's duration of response, this case suggests that combination therapy of rituximab and corticosteroids may be a promising treatment for refractory chronic ITP.

Disclosures:

No relevant conflicts of interest to declare.
Analysis of Spanish Experience During Imiglucerase Shortage
Authors: Giraldo Pilar , Irún Pilar , Alfonso Pilar , Dalmau Jaime , Fernández-Galán MAngeles , Figueredo Antonio , Ibañez Angela , Hernández-Rivas Jesús M , Luño Elisa , Marín-Jimenez Francisca , Martin-Nuñez Guillermo , Montserrat Jorge , De la Serna Javier , Villalon Lucia , Pocovi Miguel
Published: October 2019

Abstract 4725

In the last twenty years enzyme replacement therapy (ERT) with imiglucerase has been a clinically effective for Gaucher disease (GD). The recombinant glucocerebrosidase administered intravenously - usually at biweekly intervals by lifelong has improved the quality of life of patients, avoided spleen removal and bone complications. In the last months an acute shortage of imiglucerase manufactured by the Genzyme Corporation (MA, USA) has occurred as a result of viral contamination firstly and other deficiencies in the production facility. In September 2009 a position statement based on the findings of the European Working Group for Gaucher Disease and European Gaucher Alliance, established a set of key recommendations about identification and monitoring of at-risk patients threatened. In Spain the follow-up of patients and the strict complementation of rules of therapy have permitted to obtain a profile of the situation in a group of patients with restricted ERT. Patients and Methods: A total of 50 GD1 patients have been analyzed before and after 6 and 12 months of imiglucerase shortage. Have been excluded for analysis children in order to dose reduction has been minimal as well as patients who have switched to another ERT or miglustat therapy. Results: Gender: 25 males/25 females. Mean age of group: 45.3±15.3 (range:18-84) SSI at diagnosis(Dx): 8.7±3.8 (range:3-19) Chitotriosidase (CT) activity at Dx:13,383±12,783 nM/mL.h; CCL18/PARC at Dx: 767±1,198 ng/mL. 20% of patients were splenectomized and 78% had bone disease at Dx. During shortage 23 patients (46%) discontinued therapy, in this period only one patient suffered a bone crisis and other anaemia as complications. Mean reduction of haemoglobin level: 2.7% (NS), platelet counts: 5.4% (NS). CT activity was increased 135% (p<0.03) and CCL18/PARC 8.2% (p<0.08). In 17 patients (34%) imiglucerase was reduced at 50%, in this period seven patients (41.0%) suffered a bone pain and four patients (23.5%) required support therapy. Mean reduction of haemoglobin level 2.9% (NS), no changes in platelet counts. CT activity increase 48.2% (p<0.03) and not changes in CCL18/PARC concentration was observed. In addition in 3 patients (6%) the reduction was 75% and 7 patients (14%) switched to another ERT (4 patients) or miglustat (3 patients). Conclusions: In our experience, the shortage of imiglucerase in the last months has produced a incidence in bone pain of 20% and one case with anaemia and significant increase of CT activity and no significant changes in blood counts and CCL concentration, 14 % patients has required switch another therapy.

Disclosures:

Off Label Use: Lenalidomide is not approved for the treatment of smoldering multiple myeloma.
The Impact of Distance to Treatment Center on the Outcome of AML
Authors: Medeiros Bruno C. , Dunn Tamara J. , Kohrt Holbrook E , Coutre Steven , Gotlib Jason , Liedtke Michaela , Berube Caroline , Alizadeh Ash A.
Published: October 2019

Abstract 4742

Introduction

While the distance patients travel to a treatment center (DTC) adversely impacts survival of patients with trauma, cardiac, or neurological disorders, as well as certain solid tumors, less is known of its influence in acute myeloid leukemia (AML). Care for patients with AML involves frequent emergent and urgent management, often complicating primary therapy provided in distant tertiary referral centers. We therefore hypothesized that increased DTC has a negative impact on outcome. We tested this hypothesis by assessing the effect of DTC on survival of patients with AML receiving care at a single institution.

Patients and Methods

Within the Stanford Leukemia Database, we identified 884 consecutive adult patients between 1993 and 2009 meeting the following criteria: age >=18, newly diagnosed AML (excluding APL), clinical management at Stanford University Medical Center (SUMC), and verified residence location available for DTC determination. Of these, 571 were deemed fit by the admitting physician to receive myelosuppressive induction chemotherapy. DTC was calculated by straight-line journey distance between home address at the time of diagnosis and treatment center.

Results

The median age for the entire cohort is 55 years and 322 patients (36%) are older than 60 years of age. Median survival for the entire cohort was 14.0 months. DTC was not univariately associated with outcome as a continuous variable. When testing for a critical DTC threshold impacting outcomes across the entire cohort, we found a significant correlation between longer DTC and adverse outcomes, shorter DTC was associated with lower OS. Patients living within 20 miles of SUMC had a worse median overall survival (10.4 months versus 15.0 months, HR 1.23, corrected p-value 0.02). However, when adjusted for administration of induction chemotherapy (p<0.0001), age at presentation (p<0.0001) and karyotype at diagnosis (CBF vs other; p-value- 0.92), the negative impact of DTC was lost (p=0.08).

Conclusion

After accounting for confounding factors, DTC has no significant impact on the outcome of newly diagnosed AML patients receiving care at our institution. Unlike non-hematologic malignancies, distance to treatment center likely does not adversely influence outcomes for patients with AML.

Disclosures:

No relevant conflicts of interest to declare.
The Utility of the New Instrument for Comprehensive Symptom Profile Assessment In Patients with Multiple Myeloma
Authors: Novik Andrei A , Ionova Tatyana I , Kalyadina Svetlana A , Fedorenko Denis A , Mochkin Nikita E , Kurbatova Kira A , Nikitina Tatyana P , Gorodokin Gary I
Published: October 2019

Abstract 4757

Increased importance should be placed on the comprehensive symptom assessment in patients with multiple myeloma to accurately document the broad range of physical and psychological disease manifestations given from the patient perspective. The new developed symptom assessment tool - Comprehensive Symptom Profile in Multiple Myeloma Patients (CSP-MM) aims to provide an in-depth view of patient's problems. It is currently undergoing content validation through a structured, iterative process that conforms to the FDA industry guidance on the use of patient-reported outcome measures. We aimed to evaluate the utility of the new instrument providing evidence of its content validity.

Pilot sample of twenty two patients with different stages of multiple myeloma were included in this qualitative research study. Patients underwent either conventional chemotherapy or autologous stem cell transplantation. Mean age was 58 years old; male/female distribution –11/11. The CSP-MM is a self-reported tool which consists of 51numeric rating scales (where “0” - no symptom, “10” - most expressed symptom). Evidence of adequate tool content was collected from multiple sources: literature review, pilot CSP-MM completion, cognitive interviews with patients and clinicians, expert evaluation, and data quality control. Patients filled out the CSP-MM before and at different time-points of treatment.

The utility of the CSP-MM was demonstrated. After the completion of the pilot, patients were asked about their overall assessment of the questionnaire. Open-ended patient interviews provided a full understanding of the patient's perspective and showed that saturation of the items has been reached. The patients acknowledged the comprehensiveness of the tool. All of the items were easy for the patients to read and understand. Total number of completed surveys was 79 with only 1.6% missing items throughout all forms. Completion of the CSP-MM in paper and pencil format took 7–9 min. The data produced by the tool were clear for interpretation by physicians. According to the clinicians’ interviews, changes in symptom severity captured by the tool at different time-points of treatment gave the physicians information about patient experience and were used by them in day-to-day decision making.

Thus, the CSP-MM is an appropriate and practical tool to assess the symptom severity in myeloma patients. The utility of the questionnaire was shown. Pending further validation, the CSP-MM can be used to assess symptoms in MM patients and the patient benefit from the treatment.

Disclosures:

No relevant conflicts of interest to declare.
Methodological Issues In Modeling Hemophilia Treatment - What We Have Learnt In the Course of a Decade and What We Must Learn In the Decade to Come
Authors: Strandberg-Larsen Martin
Published: October 2019

Abstract 4762

Introduction:

The challenges faced by the hemophilia community because of economic constraints will increase in the future. Without disease specific pharmacoeconomic models to inform the decision process important decisions may be taken without taking hemophilia specific issues into account.

Objectives:

To investigate recent advances and current state-of-the-art knowledge in pharmacoeconomic modeling of hemophilia care we conducted a targeted literature review.

Methods:

Searches were conducted in MEDLINE®, and NHS Economic Evaluation Database using combinations of disease, and economic-related key words, limited to articles published in English between January 1970 and June 2010 (inclusive). A total of 127 articles (from 284 abstracts retrieved) were selected for detailed consideration.

Results:

Prior to the year 2000 the focus of the cost side of hemophilia treatment was not given much attention in the scientific literature. However in the last decade pharmacoeconomics have grown in importance and an increasing number of publications specific for this aspect of hemophilia reflect this trend. Still there is limited data on probabilities, costs and outcomes and remarkably few studies comparing QALY estimates of factor VIII prophylaxis in hemophilia. More studies are available specific for the limited inhibitor segment. Recommendations for reporting economic evaluations of hemophilia prophylaxis has been proposed and important ground clearing work has been published on theoretical aspects of hemophilia modeling.

Conclusion:

Economic analyses in hemophilia care are limited, but are increasingly in demand to examine the costs and outcomes associated with various treatment regimens. Unresolved questions critical to hemophilia remains such as the appropriate weighting of short term vs. future benefits and equity considerations associated with hemophilia being a rare genetic disease affecting children. Trials designed with pharmacoeconomic objectives in mind will be imperative. Data on generic and disease specific quality of life and resource use/cost data will provide whole new insights to how hemophilia impacts the lives of people living with this rare disease and will provide the data needed to assess the total value of different treatment regimens relevant for hemophilia.

Disclosures:

Strandberg-Larsen: Novo Nordisk A/S: Employment.
Urinary Tract Infection In Immunocompromised Children Under Chemotherapy
Authors: Shim Ye Jee , Lee Kun Soo , Ahn Hyo Seop , Hwang Tai Ju , Yoon Hwi-Joong
Published: October 2019

Abstract 4765

Purpose:

Infection is the most important cause of death in immunosuppressed cancer patients. And the urinary tract is a common source of infection in children. So we investigated the frequency of urinary tract infection (UTI) and the treatment outcome in children with cancer who were receiving antineoplastic therapy.

Methods:

We reviewed the medical records of children who were diagnosed as UTI during chemotherapy because of hematologic malignancies or solid tumors from January 2003 to July 2010 in Kyungpook National University Hospital. We defined as UTI when the patient showed high fever over 38.5°C and the single bacterial organism was cultured over 10,000/mL in urine sample using midstream urine collection technique. The bacterial strain, duration of fever, laboratory tests including urinalysis and gram stain, and imaging studies were demonstrated.

Results:

There were 63 cancer patients (male:female = 39:24) and 47 of them (74.6%, male:female = 30:17) experienced UTI during chemotherapy at least once. No one showed urinary symptom/sign like dysuria, frequency, urgency, flank pain or costo-vertebral angle tenderenss. The total episodes of UTI were 133 (male:female = 96:37) and the number of infection was mean 2.8 (1~10) per one patient. The common organism was Escherichia coli (25.6%), Enterococcus faecalis (15.0%), Klebsiella pneumoniae (10.5%), Enterococcus faecium (6.8%), Proteus mirabilis (6.8%) and Stenotrophomonas maltophilia (5.3%). Initial urinalysis was performed in 115 cases, but only 4 of them (3.5%) revealed pyuria. All gram stain results were negative. Duration of fever was mean 2.1 (1~6) days. The initial absolute neutrophil count (ANC) was average 1,930/μ L (0 ~ 12,610/μ L). The renal cortex scan using dimercaptosuccinic acid was performed for 43 cases to verify pyelonephritis. One showed decreased tracer uptake of upper pole of right kidney, and another 3 revealed diffuse decreased tracer uptake in both kidneys without specific photon defect. One episode of 3 was considered as urosepsis because the same organism (Klebsiella pneumoniae) was cultured from not only urine but also blood. The patient revealed hypotension and decreased renal function (glomerular filtration rate 41.4 mL/min). No mortality was observed.

Conclusion:

UTI is a very common infection in immunocompromised cancer children regardless of ANC which showed excellent prognosis with broad spectrum antibiotics. The bacterial strain was not different from that of immunocompetent children. But their symptom/sign was silent and the initial urinalysis and gram stain were not much help. So it is important to check urine culture for febrile cancer children under chemotherapy although the complication is very rare after UTI.

Disclosures:

No relevant conflicts of interest to declare.
Carbamazepine Is a Radioprotector and Radiation Damage Mitigator for Murine Hematopoietic Cell Line 32D Cl 3
Authors: Flickinger John , Shields Donna , Epperly Michael W , Zhang Xichen , Greenberger Joel S.
Published: October 2019

Abstract 4772

The search for new radiation mitigators includes targeted new- indication testing of known drugs. Carbamazepine (cbz) has been used for decades in the treatment of trigeminal neuralgia, epilepsy, mania and bipolar disorders, and has recently been demonstrated to reduce liver damage in a mouse model of alpha-1-antitrypsin deficiency by a mechanism that stimulates the proteasomal and autophagic pathway allowing degradation and clearing of abnormally folded antitrypsin. Because of its potential mitochondrial membrane ion channel effects, we tested cbz action on the irradiation survival of murine hematopoietic progenitor 32D cl 3 cells that were incubated for 1 hour in 1, 10 or 100 μ M cbz either before or after irradiation. Cells were irradiated to doses ranging from 0 to 8 Gy, plated in 4% methylcellulose, incubated for seven days at 37°C, and colonies of greater than 50 cells counted. Data was analyzed using linear quadratic or single-hit, multi-target models. Cbz added before irradiation resulted in radioprotection by an increased Do of 2.1 + 0.1 Gy (p = 0.0117), 2.3 + 0.1 Gy (p = 0.010), or 3.6 + 0.7 Gy (p = 0.0026) for 1, 10 or 100 μ M, respectively, compared to 1.5 + 0.1 Gy for control irradiated 32D cl 3 cells, with no significant change in the ñ (shoulder on the survival curve). Cbz added after irradiation mitigated against damage by an increase in the shoulder (ñ) on the survival curve with no significant change in the Do. Control irradiated 32D cl 3 cells had a ñ of 1.4 + 0.3 compared to 10.1 + 4.2 (p = 0.0113), 5.5 + 1.7 (p = 0.0185) or 3.6 + 0.8 (p = 0.0144) for cells treated after irradiation with 1, 10 or 100 μ M cbz, respectively. Cbz is known to stabilize ion channels in both cellular and mitochondrial membranes. Since irradiation induced apoptosis involves influx of calcium across the mitochondrial membrane followed by induction of nitric oxide synthase, nitric oxide and peroxynitrite formation, destabilization of the mitochondrial membrane and cytoplasmic release of cytochrome C; cbz-stabilization of mitochondrial ion channels may be the mechanism of this novel radiation mitigative effect of a commonly used drug.

Supported by NIAID grant U19AI068021.

Disclosures:

No relevant conflicts of interest to declare.
Characterization of Histone Modifications on Lineage-Affiliated Genes During Hematopoietic Stem Cell Differentiation
Authors: Fangping Chen , Tang Huarong
Published: October 2019

Abstract 4784

Hematopoietic stem cells (HSCs) are multipotent stem cells capable of self-renewal and multi-lineage differentiation. Though it has been shown that multiple factors take part in the maintenance of HSCs’ multipotency and differentiation potential, the mechanisms are unclear. Recent studies showed that histone modifications play an important role in maintenance of embryonic stem cells pluripotency and differentiation. To characterize the histone modification patterns of different lineages, HSCs were collected from umbilical cord blood and induced to differentiate to granulocytic, erythroid, and megakarytic in vitro. genes during HSC differentiation. Chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) technology was adopted to investigate the dynamic changes of histone modifications on lineage specific transcription factors and lineage–affiliated genes. Our results showed a certain level of H4 acetylation and H3 acetylation together with high level of H3K4me2 and low level of H3K4me3, H3K9me3 and H3K27me3 were present in lineage specific genes in CD34+CD38- HSCs. As CD34+CD38- cells differentiated, the modification level of acH3, acH4, H3K4me2, H3K9me3 and H3K27me3 on lineage specific genes remained the same, while H3K4me3 level increased greatly. In non-lineage specific genes, the acH3 and acH4 levels decreased, and H3K4me3 level remain at low level, while H3K9me3 and H3K27me3 levels increased. Thus, our data suggested that histone modifications played an important role in maintenaning the multipotency and differentiation capability of hematopoietic stem cells.

Disclosures:

No relevant conflicts of interest to declare.
Iinsulin Levels and Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) May Explain a Possible Important Role of the Pancreas In Sickle Cell Disease Patients Outcome
Authors: Seixas Magda , Lopes Valma , Cezar Rodrigo , Carvalho Gildasio , Rocha Larissa , Lyra Isa , Reis Mitermayer , Goncalves Marilda
Published: October 2019

Abstract 4811

Sickle cell disease (SCD) is a hematological disorder characterized by the S hemoglobin homozygosis or the association with others variants of hemoglobin. The SCD is characterized by heterogeneous clinical outcome compound by hemolysis, chronic inflammation and vaso-oclusive and pain episodes. Red Blood cell (RBC) count, hemoglobin concentration (HB), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), total bilirrubin (BT) and indirect bilirrubin (BI) are important biomarkers in SCD patients follow-up. The insulin are essential for regulation and maintenance of glucose homeostasis and the insulin resistance are associated with chronic sub-clinical inflammation and inflammatory markers such as C - reactive protein (CRP) and white blood cells (WBC) count. These markers have been correlated with the severity of SCD. The aim of this study was investigate insulin levels and HOMA-IR in SCD and it correlation with important biomarkers. Here we developed a cross sectional study involving 40 SCD patients in the steady-state of disease aging 13.9+ 8.23 years old. The RBC and WBC count were carried out by automated method and the levels of BT, BI, AST, LDH and insulin levels were investigated by immunochemistry assay method. The mean of insulin levels was 3.2 ± 2.2 and HOMA-IR was 0.61 ± 0.46. SCD patients with insulin levels lowest than the mean had an increase levels of AST (p<0.001), BT (p=0.004), BI (p=0.008), LDH (p=0.005) and WBC count (p=0.005) and decrease of HB concentration (p=0.026), HOMA-RI (p=0.001) and RBC count (p=0.006). When we studied the 25th percentile a low insulin level was correlated with high ferritin concentration (p=0.019) and in 75th percentile high insulin levels were associated with high haptoglobin concentration (p=0.024), outside difference above described. Insulin levels were negatively associated with reticulocyte (p=0.0001, r= -0.5762; BT (p=0.0003, r= -0.5444), BI (p= 0.0017, r= -0.4856) and LDH (p=0.0018 r= -0.4833). In this study the HOMA-IR was used for evaluation of insulin resistance. HOMA-IR was negatively associated with WBC count (p=0.0039, r= -0.4465); reticulocyte count (p=0.0002, r= -0.5582); BT (p= 0.0012, r= -0.4885); BI (p=0.0101, r= -0.4074) e LDH (p=0.0094 r= -0.4108. Ours results demonstrate that levels of insulin and of HOMA-IR can be influenced by the oxidative stress and inflammation secondarily of the intravascular and extravascular hemolysis in sickle cell disease, suggesting an important role of these biomarkers in the disease pathogenesis. Additional studies are warranted to explain the contribution of the pancreas in the complex net of biomarkers alteration in the disease.

Disclosures:

No relevant conflicts of interest to declare.
The Value of Beta2 Microglobulin, Retinol Binding Protein, Endothelin-1, N Amino ß-d-Glucoaminodase and Cystatin C for the Evaluation Sickle Cell Nephropathy
Authors: Unal Selma , Kotan Caglar , Delibas Ali , Arslankoylu Ali Ertug
Published: October 2019

Abstract 4817

Introduction:

Sickle Cell Anemia (SCA) is a chronic disease characterized by broad clinical manifestations and multi-organ disease. Sickle cell nephropathy is one of the major side effects in the patients with SCA. This study was planned to determine the incidence of renal complications of the patients with SCA and to evaluate the renal complications by using new markers such as ß2 microglobulin, retinol binding protein (RBP), endothelin-1, N amino ß-d-glucoaminodase (NAG) and cystatin C.

Methods:

45 children (mean age 8.4 ± 1.21, range: 1–15 years), 10 adult patients with SCA (mean age:31.4 ± 10.11, range: 16–45 years) and 20 healthy children (mean age:8.45 ± 4.05, range: 1–15 years), 10 healthy adults (mean age: 26.6 ± 8.62, range: 16–45 years) as control groups were included in the study. A detailed history was taken from each child, and a full physical examination was performed. Serum and urine electrolytes, urine analysis, urine culture and renal ultrasonography were performed for all patients. 24 hour urine was collected both in patient and control groups, and GFR was measured. Tubular phosphate reabsorbtion, 24 hour urine protein and calcium excretion, fractional excretion of Na, K, Urea and Cl were calculated from 24 hour urine samples. ß2 microglobulin, endothelin-1 RBP and NAG levels were measured in spot urine samples of patients and controls. Serum cystatin C level was measured in both patient and control groups. ß2 microglobulin, endothelin-1, RBP and cystatin C were measured with micro-elisa method. On the other hand colorimetric assay was used to measure NAG.

Results:

35 patients (63%) had enuresis nocturna, 34 patients (61%) had moderate proteinuria and 9 patient (16%) had poliuria. 26 patients (47%) had past history of urinary tract infection and 3 patients had past history of hematuria. Although all the markers to evaluate tubular functions were in normal ranges, ß2 microglobulin/creatinine ratio was found to be high in 10 patients (18%). GFR measured with creatinine was in normal ranges in all of the patients. Hovewer GFR measured with cystatin-C was low in 3 patients (0.5%). Urine ß2 microglobulin, endothelin-1, RBP, NAG, and serum cystatin-C levels were all in normal ranges both in patient and control groups.

Conclusions:

The commonly observed renal side effects in the patients with SCA are enuresis nocturna, proteinuria and increased tendency to urinary tract infections. It could be suspected that ß2 microglobulin/creatinin ratio and GFR that is calculated by the use of cystatin C could be use in order to evaluate the tubular and glomerular functions in those patients with SCA during the early stages of sickle cell nephropathy. Besides, urine endothelin-1, RBP and NAG levels could not be considered as good markers for determining the sickle cell nephropathy.

Disclosures:

No relevant conflicts of interest to declare.
Distinctive Hematological Abnormalities and GATA1 Mutation Spectra In East Asian Neonates and Children with Down Syndrome
Authors: Kim Hye-Ran , Kim Da-Woon , Baek Hee-Jo , Kook Hoon , Hwang Tai- Ju , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook , Shin Myung-Geun
Published: October 2019

Abstract 4837

Background:

Many hematological abnormalities have been reported during the neonatal period in infants with Down syndrome (DS). Neonates with DS are predisposed to developing transient abnormal myelopoiesis (TAM) and acute megakaryblastic leukemia (AMKL). However, there is a paucity of data on hematological aberrations and GATA1 gene mutations in neonates with DS in East Asian populations. We therefore investigated the spectrum of hematological abnormalities in infants and children with DS in one such population, and analyzed the frequency and pathophysiological role of GATA1 mutations in DS-TAM and DS-AMKL in this group.

Patients and Methods:

A total 148 patients with DS were identified from January 2000 to December 2009. Of these, 109 had had blood drawn for a complete blood count (CBC) with differential. There were 37 neonates (°Â4 weeks of age), 55 children (<18 years of age), and 17 adults °Ã 18 years of age) in the study. A comprehensive molecular study of the GATA1 gene was performed using PCR, RT-PCR and direct sequencing for all six exons (7,757 bp) and cDNA sequence of GATA1 gene.

Results:

Thrombocytopenia was most common hematological abnormality (35%) in the neonates. Leukocytosis and polycythemia were each detected in 4 cases (11%). Blasts were identified in the differential count of 6 (16%) neonates. Of the 55 children with DS, 11 (20%) had thrombocytopenia, 10 (18%) had anemia, and 7 (13%) had neutropenia (< 1,500/uL). There were also 4 (7%) cases of AMKL and 2 (4%) cases of acute lymphoblastic leukemia in the children with DS. GATA1 mutations were identified in all of the TAM samples and in 2 of the 3 AMKL samples, but were not detected in the 1 remission sample or the 2 samples from normal DS children.

Conclusions:

East Asian DS neonates showed distinctive hematological abnormalities such as uncommon neutrophilia, relatively common neutropenia, and increased prevalence of TAM compared with reports from Western countries. The GATA1 mutation analysis may offer the potential to accelerate and refine the diagnosis of TAM and AMKL, and to accurately measure the kinetics of the mutant GATA1 clone as a tool for minimal residual disease measurement because virtually all cases of TAM and AMKL harbored this mutation.

Disclosures:

No relevant conflicts of interest to declare.
P53 Abnormality and Prognostic Impact In Leukemia Patients with Complex Chromosomal Abnormalities
Authors: Cao Xiangshan , Zhou Zhaoling
Published: October 2019

Abstract 4847

Objective:

To investigate the prevalence of p53 gene deletion in leukemia patients with complex chromosome abnormalities (CCA) and their relative prognostic impact.

Methods:

The chromosome abnormalities in acute leukemia cases with R banding method, karyotype is named according to international nomenclature ISCN 2005. Interphase fluorescence in situ hybridization (I-FISH) determinations were performed to detect the percentage and frequency of p53 gene deletion in 38 leukemia patients with CCA and 24 patients without CCA. The rate of complete remission (CR) and median survival time (MST) were observed between the two groups in patients with CCA or without CCA.

Results:

(1) Complex karyotype abnormality in the leukemia patients involved in almost all of chromosomes, especially chromosome 17, chromosome 5, and chromosome 7. And the reduction of chromosome number was more common than the increase. (2) The threshold of two green and one red (2G1R) was 7%, the threshold of one green and one red (1G1R) was 4.9%. (3) The percentage of p53 gene deletion was 44.74% in CCA group and 4.16% in non-CCA group (p<0.0001). (4) p53 gene deletions were found in 9 of 10 cases with chromosome 17 abnormalities. The percentage of p53 gene deletion was achieved in 90%. (5) The CR rate was 57.14% and the MST was 31 months in patients with non-CCA compared to 11.76% CR rate and 2 months MST in patients with CCA (p<0.0001).

Conclusion:

The deletion of p53 gene appears to be associated with complex chromosome abnormalities and may play an important role in the prognosis of patients with leukemia

Disclosures:

No relevant conflicts of interest to declare.
Frequency of Citogenetic Abnormalities and the BCR/ABL Fusion Gene In Adult Patients with Acute Lymphoblastic Leukemia (ALL). Multicentric Study From Mexico
Authors: Ignacio Gregorio , Arana Rosa_Maria , Solis Juan_Carlos , Ramos Christian , Gonzalez Mauricio , Perez Oscar , Jimenez Rosa_Maria , Lugo Yolanda , Aguilar Lilia , DeDiego Jope , Guzman Laura , Rozen Etha , Limon Alejandro , Lopez Manuel , Nacho Kevin
Published: October 2019

Abstract 4864

The t(9;22) or BCR/ABL fusion genes occurs in 20–30% of adult patients with ALL overall and its presence leads to worse prognosis for ALL. But there are a few large reports focused on this subtype of adult ALL in México, including its clinical characteristics.

Objective.

(1) To know the frequency of chromosome Ph-positive or BCR/ABL by cytogenetic and RT-PCR studies; (2) to evaluate the clinical prognostic factors and the correlation with cytogenetic abnormalities.

Methods.

In the present report, we study of 104 adults with ALL de novo from seven different hospitals from Mexico, between November 2008 – June 2010. All patients were de novo and were subjected to karyotyping in bone marrow and RT-PCR from BCR/ABL at diagnosis.

Results.

From 104 ALL adults included 47(45%) males and 57(54.8%) females, with age range 17–73 years (mean= 33.81 years). The most frequent subtype FAB were L2 (87%) an L1 (13%) with immunophenotype by LAL-B 95%, and 4.8% bilinear. Blood counts were very low, mean hemoglobin 7.8 g/dL (SD+ 2.61), platelet 67,266 × mm3 (SD 79,627), and leukocytic 37,815 × mm3 (SD 60,905). The grown of liver, spleen and adenomegaly was only 40.4%. 19 (18.26%) karyotypes was not useful. Eigthfive informative karyotypes were subdivided into 6(7%) t(9;22); 49(57.6%) normal; 14(16.5%) hyperdiploid; 2(1.9%) hypodiploid; 2(1.9%) poliploid; and 5(5.8%) rearr(12). Chromosomes 9,11,12 and 3 were the most frequently envolved by structural rearrangements. The frequencies of the BCR/ABL studied with RT-PCR (minor and major break point region); only 4 cases was not useful, then 20/100 was positive; only 2/20 (10%) with the b2a2/b3a2 transcript of the P210BCR/ABL; the rest 90% with e1a2+ of the P190BCR/ABL.

Conclusion:

The frequency reported for BCR/ABL positive (20%) in Mexican adults with ALL is in concordant with worldwide; but only by RT-PCR. The karyotype has low sensitivity by detection of the t(9;2) but is useful by identify others anormalities in karyotype. We don't have correlation between clinical and cytogenetic characteristics.

Disclosures:

No relevant conflicts of interest to declare.
Invasive Mould Infections (IMI) In Acute Leukemia Patients on Chemotherapy - Can We Win the Battle?
Authors: Wong Gee Chuan , Tan Ban Hock
Published: October 2019

Abstract 4871

Aim: To determine risk factors for, and clinical features of, invasive mould infections in patients with acute leukemia on chemotherapy.

Methods: A matched case control study where acute leukemia patients on chemotherapy were identified and clinical data pertaining to their disease, clinical course, IMI characteristics and treatment outcomes were evaluated. This was conducted in a tertiary hospital from 1 January 2004 to 31st March 2007.

Results: All acute leukemia patients on chemotherapy receive itraconazole prophylaxis. Nineteen cases of invasive mould infections (IMI) were found in the period, giving an incidence of 11.2%, using only patients undergoing curative chemotherapy as denominator. The incidence in patients receiving the first cycle of chemotherapy was 8.9%; in patients receiving subsequent cycles of chemotherapy was 4%. Thirteen cases were proven by EORTC/MSG criteria, 8 of them by histology without positive cultures. There were 3 proven cases of fusariosis, and 1 each of aspergillosis and nodulisporiosis. Cases were more likely than controls to have had an absolute neutropenia for at least 14 days in the chemotherapy episode in which the IMI was diagnosed (p=0.045), to have been bacteremic prior to the diagnosis of IMI (p=0.045), and to have had fever not responding to a carbapenem (p=0.016). They also had a longer length of stay (51.8 vs 27.2 days, p=0.01). The use of itraconazole as anti-fungal prophylaxis was not less common in cases than controls. Common clinical features noted in cases included fever not responding to a carbapenem (84.7%), cough (42.1%), a rise in alkaline phosphatase (36.9%), breathlessness (26.3%), and hemoptysis (21.1%). A pleural effusion was noted on CXR or CT chest in 52.8% of cases, and a characteristic CT thorax in 47.4% of cases. Sixty-eight percent of cases received amphotericin (conventional or lipid preparation) as part of a febrile neutropenia protocol, though only 15.8% received this agent when an IMI was considered. Clinical improvement was attributed to an increase in ANC in 26.3% of cases, and to the introduction of voriconazole in 10.5%. The next cycle of chemotherapy was delayed for 2 – 4 weeks in 15.8% of cases, and for >4 weeks in 31/6% of cases. Three patients (15.8%) died within three months of IMI diagnosis, 47.4% survived >1 year.

Conclusions: Anti-fungal prophylaxis with itraconazole did not reduce the likelihood of IMI and this should be changed. Absolute neutropenia is a risk factor for IMI in patients with acute leukemia. Fever not responding to carbapenems and a rise in ALP may be considered as red flags for clinicians to consider the possibility of an IMI and institute prompt treatment.

Disclosures:

No relevant conflicts of interest to declare.
Essential Role of γδT Cells In Naturally Acquired Immunity to Falciparum Malaria
Authors: Kanda-Taniguchi Tomoyo , Mannoor Kaiissar Md. , Li Changchun , Watanabe Norihiro , Yamahira Akie , Phompida Samlane , Narita Miwako , Watanabe Hisami , Takahashi Masuhiro
Published: October 2019

Abstract 4877

It is important to understand the mechanisms of naturally acquired immunity to malaria for the development of effective malaria vaccines. We have demonstrated that γδT cells expanded in the peripheral blood of the falciparum malaria patients in Thailand but did not expand in patients living in malaria endemic areas of Laos. However, the percentage of Vγ9-T cells, a γδT cell subset, increased in the Laos patients. The levels of naturally acquired antibodies to crude P. falciparum (Pf) antigens also increased with an age dependent manner in individuals living in endemic areas of Laos. In this study, we further investigated the role of Vγ9-T cells in naturally acquired immunity to the falciparum malaria. The peripheral blood lymphocytes (PBLs) and plasma obtained from hospitalized uncomplicated falciparum malaria patients (UMPs) and severe falciparum malaria patients (SMPs) in Thailand and from non-hospitalized UMPs living in endemic areas of Laos were analyzed. The plasma levels of IL-10, which is anti-inflammatory cytokine and associated with antibody production from B cells, were elevated in both hospitalized and non-hospitalized falciparum malaria patients. There was a correlation between the levels of IL-10 and the percentage of Vγ9-T cells in γδT cells. IL-10 and Pf specific antibodies were detected only in culture supernatant of PBLs from non-hospitalized UMPs in the presence of IL-2 for 2 wks. These results indicate that Vγ9-T cells may contribute to acquiring natural immunity to malaria.

Disclosures:

No relevant conflicts of interest to declare.