bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of GATA1 Mutation in the Blood of 3 Mothers Giving Birth to Children with Trisomie 21 and TMD.
Authors: Kabisch Hartmut , Obser Tobias , Bartl Sonja , Kordes Uwe , Blohm Martin , Schneppenheim Reinhard 
Published: October 2019

Abstract
Abstract 4680
One of the most common human chromosomal abnormalities, Down syndrome, is caused by trisomy 21. Transient myeloproloferative disorders (TMD) occur in 10-20 % of newborns with Down syndrome. About 30 % of those TMD patients develop Acute Megakaryoblastic Leukemia (AMKL) within 4 years. Therefore both disorders have their origin in the fetus. There is emerging evidence that the protein product of GATA1 mutations, GATA1s, directly contributes to leukemogenesis. Although an in utero origin of GATA1 mutations and consecutively TMD is established it was never analyzed if GATA1 mutations could be detected in maternal PBL collected before and after the delivery of a child with Down syndrome and TMD. We therefore initiated a study to detect mutant GATA1 in three pairs of mother and child (A, B, C).
After the identification of the specific GATA1 mutations of the newborn with TMD, PBL samples of the mothers drawn 4 days after delivery (mother A), 7 days after delivery (mother B) and 4 week before as well as just before sectio cesarea ( mother C) were analyzed.

Results
All 3 patients had duplications in exon 2. Patient A had a duplication of 191 bp including the start codon,, patient B showed a duplication of 2 bp, and patient C had a duplication of 13 bp, respectively.
The three mothers harbored the TMD specific GATA1 mutations in PBL analyzed around the birth of their children.
The GATA1s signal in mother A and B had disappeared 1 month after birth. No analysis at this time point was done in mother C,whereas she was positive for the specific GATA1 mutation one month before delivery. At that time TMD was suggested by ultrasound which revealed clinical signs of hydrops in the fetus.
In mother B and C the particular GATA1 mutations could also be detected in their plasma, in the plasma sample of mother C even 4 weeks before delivery (not done in mother A).

Conclusion
In our study we could show for the first time that GATA1 mutations of leukemic cells could be detected in PBL of the mothers around birth. Cell free GATA1 DNA was also detectable. The prenatal finding of TMD specific mutations in the mothers' blood suggests that feto-maternal transfer of leukemic cells and DNA may already occur up to 4 weeks before delivery.

Disclosures:
No relevant conflicts of interest to declare.
P15 and P16 Promoters Methylation Status in APL Patients Treated by Arsenic Trioxide.
Authors: Yeganeh Omid , Alimoghaddam Kamran , Ghaffari Seyed Hamidolah , Rostami Shahrbano , Jalili Mahdi , Shamshiri Ahmad Reza , Ghavamzadeh Ardeshir 
Published: October 2019

Abstract
Abstract 4682

Introduction
Inactivation of tumor suppressor genes P15 & P16 due to Hypermethylation has a critical role in progression of cancer. We analyzed the meltylation status of promoter region of these two genes and their correlation with patients survival in APL patients treated with ATO without chemotherapy and /or ATRA.

Methods
Patients treated by Arsenic Trioxide without ATRA or chemotherapy and 45 newly diagnosed and 5 relapsed were enrolled in our study. Whole blood DNA was extracted and modified by sodium bisulphite reaction. Then methylation status was studied by MS-PCR and gel electrophoresis. Also for newly diagnosed patients, early mortality, disease-free and overall survival compared between methylated or non-methylated groups.

Results
Of 45 new cases, 31patients(69%) had methylated P15 but no patient had methylated P16. Eighty percent of relapsed patients had methylated P15 too, and no relapsed patient had methylated P16. DFS and OS correlated significantly with methylation status of P15 ( P=0.05 and 0.04) for patients who achieve to CR after treatment but it has no correlation with early motility during treatment course. Actually methyaltion of P15 improve chance of DFS and OS.

Conclusion
P15 but not P16 is frequently methylated in APL. Although in some studies, methyaltion of P15 negatively affected prognosis of APL patients treated by ATRA but it has no adverse effect when Arsenic Trioxide used in induction and cosolidation.

Disclosures:
No relevant conflicts of interest to declare.
Familial Platelet Disorder and Predisposition to Myeloid Leukemia (FPD/AML) in the Absence of RUNX1 Mutations- A Report of Three Families.
Authors: Berman Jason N. , Couban Stephen , Ebbin Allan , Frangoul Haydar , Quisumbing Maria B. , Aikie Thomas E , Riddell Christie , Ferguson Meghan , Ludman Mark D. , Fernandez Conrad V. , Cantor Alan B. 
Published: October 2019

Abstract
Abstract 4689
Familial Platelet Disorder with Predisposition to Acute Myeloid Leukemia (FPD/AML) has been attributed to abnormalities of the RUNX1 gene, located at human chromosome 21q22. The protein product of this gene forms a critical component of the core binding factor (CBF) complex, which is required for normal hematopoiesis. Truncating mutations and deletions of RUNX1 (particularly in exons 3, 4, and 5, encoding the DNA binding and CBF-beta interaction domains) that result in haploinsufficiency or dominant-negative activity have been identified as the genetic mechanisms underlying FPD/AML. Clinically, these patients present with mild to moderate thrombocytopenia, platelet dysfunction, bleeding, and about a 35% risk of developing AML. We report on three unrelated families who share histories of autosomal dominant thrombocytopenia over two to five generations, bleeding, platelet aggregation defects, and development of myelodysplastic syndrome, AML or intriguingly, chronic myeloid leukemia (CML). Detailed genetic interrogation of the RUNX1 locus in the proband from each family, including complete sequencing of all 8 exons, flanking 50 base pair regions, and P1 and P2 promoters, as well as gene dosage studies, failed to demonstrate a causative lesion in the RUNX1 gene. Our findings strongly suggest that genetic loci other than RUNX1 are involved in some cases of autosomal dominant thrombocytopenia with a predisposition to both AML and CML. Identification of the disease-causing genes in these families will allow for prospective testing of family members, appropriate surveillance and early intervention in affected individuals, and potentially new molecular insights into leukemogenesis. We have broadened the genotype-phenotype correlation in FPD/AML beyond the RUNX1 gene and suggest that this syndrome may be more genetically heterogeneous than initially described.

Disclosures:
No relevant conflicts of interest to declare.
Loss of Structural Maintenance of Chromosomes 1A Protein Expression Is Associated with a Poor Prognosis in Acute Myelogenous Leukemia.
Authors: Krug Utz O. , Hömme Claudia , Tidow Nicola , Buerger Horst , Koehler Gabriele , Heinecke Achim , Buchner Thomas , Berdel Wolfgang E , Koschmieder Steffen , Mueller-Tidow Carsten 
Published: October 2019

Abstract
Abstract 4693
Loss of Structural Maintenance of Chromosomes 1A Protein Expression is Associated with a Poor Prognosis in Acute Myeloid Leukemia
Utz Krug, Claudia Hömme, Nicola Tidow, Horst Bürger, Gabriele Köhler, Achim Heinecke, Thomas Büchner, Wolfgang E. Berdel, Steffen Koschmieder, Carsten Müller-Tidow

Introduction
Acute myelogenous leukemia is a genetically heterogenous disease with many risk factors for a poor prognosis. One of the most important independent risk factor in AML is the age at diagnosis. Older patients with AML generally have a poor prognosis which suggests that the biology of AML in elderly patients differs from AML in younger patients.

Methods
Gene expression profiling was carried out to identify age related changes in AML blasts of 67 AML patients of different age (range: 17 to 80 years). Among the genes that correlated with age, SMC1A was selected for protein expression studies. A tissue array was created containing bone marrow histology samples of 135 patients with newly diagnosed AML of different ages and probed with an antibody against SMC1A and protein expression was quantified by the DAKO score.

Results
131 genes showed a significant correlation between mRNA expression levels and patient age. Increasing age was associated with significantly decreased mRNA levels of SMC1A. 116 patient samples were evaluable for SMC1A protein expression and expression of SMC1A protein was low or absent in 74 out of 116 AML specimens. SMC1A protein expression did not show a correlation with patients' age at diagnosis. Both event free survival (2.6 months vs. 10.3 months, p=0.003, see figure) and overall survival (10.4 months vs. 22.6 months, p=0.015, see figure) were significantly worse in patients with low or absent SMC1A protein expression. In a multivariate analysis, SMC1A protein expression level remained a significant prognostic factor for event free survival (p=0.014) with a borderline significance for overall survival (p=0.066).

Conclusions
We identified 131 genes with putative age-dependent microarray mRNA expression and identified low levels of SMC1A protein expression as a marker for poor prognosis in patients with newly diagnosed acute myeloid leukemia.

Disclosures:
No relevant conflicts of interest to declare.
Detection of NFκB Activity in Primary CLL Cells Using a Sensitive Oligo-Based Chemiluminescent ELISA.
Authors: Mertens Daniel , Bhattacharya Nupur , Sarno Antonio , Idler Irina , Zenz Thorsten , Döhner Hartmut , Stilgenbauer Stephan 
Published: October 2019

Abstract
Abstract 4709
Contemporary research on cellular signaling in oncogenesishas undergone a shift of focus from qualitative measurements of single signaling pathways to high-throughput quantitation of comprehensive signaling networks. Notably, NFkB is a family of transcription factors involved in immune and inflammatory responses, developmental processes and especially in cellular growth and apoptosis, which are central in oncogenesis: NFkB signaling is deregulated in a number of hematologic malignancies like Hodgkin's lymphoma or chronic lymphocytic leukemia. We have established a chemiluminescent oligonucleotide-based ELISA (co-ELISA) that is simple and quantitative. In contrast to currently used assays, it allows quantitation of all NFkB components (i.e. RelA, p50, p52, RelB, and c-Rel). In addition, it can make use of whole extract and does not require cumbersome nuclear/cytosolic fractionation, saving time and resources. Co-ELISA has a 3.5- to 43-fold higher signal-over-noise ratio than currently available assays, while the standard error (SEM) is 3-to 6-fold lower. Co-ELISA is suited for the assessment of NFkB activity in primary tumor cells or in high-throughput experimentation i.e. functional evaluation of novel compounds. The method is faster than EMSA, not restricted to transfectable cells as is the case for luciferase reporter assays, making it applicable to primary tumor cells, and ten times more cost-efficient than commercially available ELISA assays. Co-ELISA is a quantitative, sensitive, fast, and cost-efficient measurement of the activity of all DNA-binding NFkB proteins, also in primary tumor cells.

Disclosures:
No relevant conflicts of interest to declare.
Expression of KDR Plus Detection of Flt3-ITD and NPM1 Mutation as a Prognostic Marker in AML Patients with Normal Karyotype.
Authors: Liang Aibin , Xiu Bing , Huang Binbin , Han Ying , Bo Lanjun 
Published: October 2019

Abstract
Abstract 4715
From July 2006 to February 2009, total of 146 patients with non-APL AML were admitted in our department. Cytogenetic analyses, detection of Flt3-ITD, NPM1 gene mutation and VEGF and its receptors (Flt-1, KDR) were performed on all patients. Forty-nine patients less than 60 years of old and with normal karyotype were selected for prognostic analyses. Flt3-ITD was positive detected in 15 cases (30.61%), NPM1 mutation in 18 cases (36.73%), VEGF in 46 cases (93.88%), Flt-1 in 41 patients (84.04%) and KDR in 38 cases (77.55%). After one or two courses of induction therapy with IDA regimen (Idarubicin + cytarabine: 3+7) in all 49 patients, total CR rate was 67.43% (33/49). One patient died because of severe invasive fungal infection. Among the remaining 15 non-CR patients, 10 were Flt3-ITD positive and NPM1 negative, and all with higher expression of VEGF and KDR. All the CR patients were treated with consolidation regimen with high dose cytarabine (3g/m2, q 12h iv for 3 consecutive days) for 6 courses. After follow-up at least for 6 months, only 12 patients are alive up to now. In these 12 patients, Flt3-ITD were all negative expressed, 6 patients were NPM1 positive, 2 patients VEGF negative, 3 patients both KDR and Flt-1 negative. There are no patients alive with positive expressed of Flt3-ITD, KDR and negative expressed of NPM1. Therefore, we supposed that negative expression of Flt3-ITD and KDR plus positive expression of NPM1 could be a favorable parameter for outcome prediction in AML patients with normal karyotype.

Disclosures:
No relevant conflicts of interest to declare.
Aven and Survivin Expression in Egyptian Acute Leukemia and Their Relation to Apoptosis.
Authors: Assem Magda , Hamid Thoraya Mohamed Abdel , Basset Gihan Abedel , Hilal Amany , Kamel Mahmoud , Khalil Iman , Alsharkaway Nahla , Metwally Aymen 
Published: October 2019

Abstract
Abstract 4717

Background
Several anti apoptotic signals have been recently identified. Aven and Survivin are broadly expressed and are conserved in mammalian species.

Patients and Methods
Sixty-four patients with acute leukemia [39 with acute myeloblastic leukemia (AML) and 25 with acute lymphoblastic leukemia (ALL)] were used as the study group. Reverse transcriptase (RT –PCR) was used, Aven and Survivin expression were detected at the messenger (mRNA) level .DNA fragmentation was carried out on daily basis before &24 hours after the therapeutic dose every day for 4 times during the initial induction of chemotherapy (everyday for ALL& every otherday for AML).

Results
Survivin expression was found in (66%) of acute leukemia, near significantly (p=0.06) more in AML (74%) than in ALL (52%). While, Aven expression was (40.6%) in acute leukemia equally expressed in AML (41%) and ALL (40%).Patients were categorized into 3 groups based on DNA fragmentation, whereby group I: DNA fragmentation found on day 1 or day 1+day 2, group II: DNA fragmentation found on days 1, 2 and 3, group III where fragmentation was found on days 1+2+3+4 or/and day5. Absence of Aven expression significantly (p<0.001) contributed to DNA fragmentation as 24 /35 (68.6%) of acute leukemia with Aven negative were in group III (the best group). Absence of Survivin did not contribute as much, Where 18 out of 31 (58%) of group III were Survivin positive (P= 0.1).None of the concordant both positive Survivin and Aven were in group III (the good 5 day fragmentation), this was statistically highly significant (P< 0.001) relation. Survivin was statistically related to CD7 expression (P<0.001) in AML only, while Aven was statistically related to CD34 expression (P<0.014) in the whole acute leukemia cases. Aven was statistically correlated to alkaline phosphatase (P< 0.036).There was a significant dissociation between Aven and Survivin in AML (p=0.03) and near significant dissociation in ALL (p=0.07).

Conclusion
Aven seems to be more important as an inhibitor of apoptosis than survivin in acute leukemia. The presence of both Aven & survivin confers a survival disadvantage & a significantly worse DNA fragmentation pattern, thus suggesting a synergistic inhibition of apoptosis when present together, although, they tend not to be expressed together. The highly significant relation between CD7 & surviving expression, might suggest their involvement in a common signal transduction pathway.
Key words: Survivin, Aven, AML, ALL and DNA fragmentation.

Disclosures:
No relevant conflicts of interest to declare.
Cytogenetic and Molecular Features of 1331 Chinese Patients with Primary Acute Lymphoblastic Leukemia (ALL).
Authors: Chen Bing , Wang Xiang , Yao Ying , Zhu Yong-Mei , He Hai-Yan , Jiang Xiao-Xing , Zhou Ji-Fang , Chen Hai-Min , Cao Qi , Yang Guang , Jiang Chun-Lei , Weng Xiang-Qin , Zhang Xu-Xi , Jiao Bo , Zhao Wei Li , Li Jun-Min , Chen Yu , Xiong Shu-Min , Shen Zhi-Xiang , Gu Long-Jun , Jiang Hui , Mi Jian-Qing , Chen Zhu , Chen Sai-Juan 
Published: October 2019

Abstract
Abstract 4718
Cytogenetic-molecular signature plays an important role in diagnosis, treatment and prognosis evaluation in acute lymphoblastic leukemia (ALL). With examination of karyotype and gene rearrangement, we investigated retrospectively the cytogenetic characteristics in 1331 adults and children with primary acute lymphoblastic leukemia (ALL) diagnosed from November 1989 to July 2007.
Cytogenetic and molecular genetic analysis was carried out successfully in 1030 (77.39%) of 1331 cases, abnormal clone was detected in 629 (61.07%) cases. The abnormalities of t(9;22)/BCR-ABL and/or t(4;11)/MLL-AF4 were presented more commonly in adults than in children (25.39% Vs 10.34%,P<0.001; 4.69% Vs 1.68%,P=0.0067). On the contrary, hyperdiploid(>50) was more frequently seen in children than in adults (13.18% Vs 4.30%,P<0.001). In addition, t(8;14)/IGH-MYC,t(12;21)/TEL-AML1 and t(11;19)/MLL-ENL were only observed in pediatric ALL. Compared with data of western countries, t(9;22)/BCR-ABL involvement in Chinese ALL patients, either in adults or in children, was more common. Simultaneously, a lower frequency of t(12;21)/TEL-AML1 was found in Chinese pediatric ALL patients.
Ik6, one of IKZF1 deletion isoforms, was detected in 72.29% of ALL patients with Ph chromosome using reverse transcription-PCR amplification.
In T-ALL, the frequency of the expression of SIL-TAL,CALM-AF10,HOX11,HOX11L2 gene and the mutation of NOTCH1 gene was 12.38%,4.69%,25.49%,24.51% and 32.99%, respectively.
The overall survival of the children was superior to that of the adults, which was the same as previous reports by many other groups. According to the prognosis of patients, these cytogenetic-molecular signatures could be further classified into three subgroups in adult and four subgroups in children. Patients with Ik6 deletion isoform of IKZF1 gene had a significantly worse prognosis than those with wild type isoform (P=0.009). In T-ALL, patients with NOTCH1 mutation were related with poor outcome (P=0.007).

Disclosures:
No relevant conflicts of interest to declare.
Investigation On Exosomes Carrying TSA Derived From Healthy Human White Buffy Coat with Positive HLA-A2 and PolyI:C On Subcellular Antitumor Vaccination.
Authors: Ren Weina , Chang Chunkang , Li Xiao 
Published: October 2019

Abstract
Abstract 4747

Aim
Use human leukocyte antigen –A2 (HLA-A2) positive human dendritic cell (DC) derived DEX to support NY-ESO-1 antigen and polyI:C to increase the proliferation of specific cytotoxic T lymphocyte (CTL) in transgenic mice, and increase its anti-tumor effect.

Methods
Mature dendritic cells derived from peripheral blood mononuclear cells (PBMC) are isolated from the blood from healthy adults with positive HLA-2A. By centrifugation and membrane ultrafiltration, EXO is extracted from the supernatant of DC secretions. Transgenic C57 mice were immunized by human derived tumor testis antigen NY-ESO-1/EXO with or without polyI:C. Mice were sacrificed four weeks after immunization, and spleen cells were isolated and underwent function test. The experiments include: antigen specific CTL proliferation efficiency was tested by dimerization experiment; the antitumor effect for K562 cells and melanoma were tested under different ratio between effecter cells and target cells (0:1, 10:1, 50:1 and 100:1).

Results
Dimerization experiment indicated that the effect of DEX/TSA+PolyI:C was (1.98±0.79)%. The effect of DEX/TSA was (1.61±0.58)%. The antitumor effect for the ratio of 0:1, 10:1, 50:1 and 100:1 by DEX/TSA:PolyI:C were (11.14±1.36%) A (14.17±0.62%) A (15.71±2.48%) A (24.31±2.91%) for K562 cells; The antitumor effect for DEX/TSA group (12.23±2.25%)A(13.10±1.57%)A(15.27±2.93%)A(19.87±2.72%)for K562 cells; With ratio 10:1 and 100:1, the antitumor effect of DEX/TSA +PolyI:C is better than DEX/TSA group (P<0.05). Whereas, with increased ratio between effecter cells and target cells, there is no significant improvement on antitumor effect for control cells.

Conclusion
It is promising to combine DEX/TSA derived from healthy human blood with positive HLA-A2 and PolyI:C as a new subcellular antitumor vaccination.

Disclosures:
No relevant conflicts of interest to declare.
Whole Brain Radiation Alone Brings the Favorable Outcomes for AIDS Related Primary Central Nervous System Lymphoma (PCNSL) Especially with Good Performance Status in the HAART Era.
Authors: Nagai Hirokazu , Odawara Takashi , Ajisawa Atsushi , Hagiwara Shotaro , Uehira Tomoko , Uchiumi Hideki , Yotsumoto Mihoko , Miyakawa Toshikazu , Watanabe Akira , Konishi Mitsuru , Saito Seiji , Takahama Souichirou , Tateyama Masao , Okada Seiji 
Published: October 2019

Abstract
Abstract 4751

Background
Primary central nervous system lymphoma (PCNSL) is one of an acquired immunodeficiency syndrome (AIDS)-defining illness and is the second common cerebral mass lesion after toxoplasmosis in HIV infected population. The introduction of highly active antiretroviral therapy (HAART) significantly improved the survival of AIDS related PCNSL. But, the standard procedure to manage these patients still remains to be established. We performed the nationwide retrospective survey to elucidate the actual status and the significant prognostic variables of AIDS-related PCNSL in the HAART era.

Methods
All AIDS related PCNSL newly diagnosed at 12 regional center hospitals for HIV/AIDS in Japan during the period 2002-2008 were registered. The data about diagnostic modalities, CD4 counts, performance status (PS), HAART regimens, treatment modalities, and clinical outcomes were collected. A multivariate and univariate Cox regression analysis was performed to assess the effects of treatment and the clinical variables on OS.

Results
Twenty four AIDS related PCNSLs were registered. Median age was 41 (21-60) year old, and male gender was 96%. Ten patients were diagnosed as PCNSL based on imaging features and positive EBV DNA PCR of cerebrospinal fluid without brain biopsy. HAART was administrated in 88%. Twenty two patients were treated by radiotherapy alone, and only one cases were received the combined modality treatment. Multivariate analysis revealed that the significant predictor of improved survival was completion of whole brain radiotherapy (WBRT) with a total dosage more than 30 Gy (P=0.017). Furthermore, 9 of 10 patients with good PS (0-2) group were alive with CR (all were received WBRT), nevertheless 64% of poor PS (3 or 4) group (9/14) died sometimes with short clinical course. The estimated 3yrs overall survival rate of each group was 100% and 38% respectively (p=0.0198). The incidence and grade of radiation related leukoencephalopathy was limited in good PS group.

Conclusion
Radiotherapy alone with HAART is shown to be the most common treatment for the AIDS-related PCNSLs, but the role of systemic chemotherapy should be elucidated. Some AIDS-related PCNSLs, especially with good PS, achieved the durable remission by WBRT with the curative intent dose, in the HAART era.

Disclosures:
No relevant conflicts of interest to declare.