bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relationship Between Intestinal Marginal Zone B-Cell Lymphoma and Enterobacterias Including Camphylobacter Jejuni
Authors: Oh Sung Yong , Kim Won Seog , Kim Seok Jin , Hong Junshik , Lee Soon Il , Lee Jae Hoon , Do Young Rok , Kim Min Kyung , Lee Ho Sup , Suh Cheolwon , Yi Seong Yoon , KIm Jong Hyun , Hong sook Hee 
Published: October 2019

Abstract
Abstract 5224

Background:
Even though one great study had been addressed for identifying relationship Camphylobacter jejuni (C.jejuni) and IPSID, association is not proof for causation; it is usually the first step in proving the microbial etiology of a disease. Intestinal-marginal zone B-cell lymphoma (I-MZL) in Korean result, many points of clinical presentations were quietly different from classical IPSID. Median age was about 60 not a young child, common involving site was ileo-cecal valve not a jejunum, limited segmental involvement was dominant not a diffuse involvement of whole intestine. Therefore, we propose this study for identifying the causative etiology of I-MZL. Thus firstly we planned to demonstrate the enterobacterias including C.jejuni and I-MZL with Korean specimen.

Patients and Methods:
Histologically confirmed 33 patients of intestinal MZL were evaluated for this study. DNA preparing from FFPE will be used Ambion RecoverAllTM Total Nucleic Acid Isolation Kit for FFPE tissue. Universal primers PB (5'TAACACATGCAAGTCGAACG3') and DG74 5'AGGAGGTGATCCAACCGCA3') has been used to amplify bacterial 16S ribosomal DNA (rDNA). Established primers directed at C. jejuni (CCCJ609F 5'AATCTAATGGCTTAACCATTA3' and CCJ1442R 5'GTAACTAGTTTAGTATTCCGG3') will be used to generate an 852-bp 16S rDNA amplification. Cycling conditions for all PCR will be as follows: 7 minutes at 95°C, 35 cycles of denaturation for 30 seconds at 95°C, annealing for 20 seconds at 55°C, and 2 minutes of extension at 72°C and then 10 minutes at 72°C.With the similar concept, Salmonella typi, Vibrio Cholerae, Escherichia coli, Shigella also were evaluated for evaluating the etiology of the I-MZL.

Result:
Between 1999 and October 2009, a total of 24 patients with histological diagnosis of I-MZL from 9 different institutions in Korea were analyzed. The male/female ratio was 14 to 10. The median age was 57 years (range, 15–80). The number of involved site was duodenum-1, jejunum- 2, ileum-7, ileo-cecal valve-3, ascending colon-2, sigmoid colon-1, rectum- 5, and multiple intestine involvement was 3. Four bands were appeared in I-MZL patient. All of two jejunum involved MZL tissue had positive result. But one patient was ileum and the other one was ileo-cecal valve involved MZL. Salmonella typi, Vibrio Cholerae, Escherichia coli, Shigella were all negative in I-MZL tissue

Conclusion:
It was a difficult to identify the relationship several enterobacterias and I-MZL. However C. jejuni could be a possibility of etiologic organism of jejunum involved MZL. Furthermore, extra-jejunum I-MZL has a need for more investigation to define the etiology of I-MZL

Disclosures:
No relevant conflicts of interest to declare.
Prox1 Suppressed Tumor Cell Growth by the Down Regulation of PKCβII Through the DNA Methylation
Authors: Hagiwara Kazumi , Murate Takashi , Miyata Yasuhiko , Hotta Tomomitsu , Nagai Hirokazu 
Published: October 2019

Abstract
Abstract 5227

BACKGROUND:
The homeobox gene Prox1 is a transcription factor related to the Drosophila prospero gene and known to be the master gene which controls the development of lymphatic vasculature. Recent studies have suggested that Prox1 is a strong candidate tumor suppressor gene in several kinds of malignancies. We previously demonstrated that the expression of Prox1 gene was frequently silenced by aberrant DNA methylation in about 60% of diffuse large B cell lymphoma (DLBCL). In this study, we showed that Prox1 regulated Protein Kinase C beta II (PKCβII) expression, which was overexpressed in poor risk DLBCL, and analyzed the inhibitory mechanism of tumor suppression by Prox1.

METHODS and RESULTS:
We established the Prox1 stable transformants of Hela cells which originally lacked Prox1 expression, and examined them in vitro cell proliferation and in vivo tumor formation using nude mice. Our experiments showed that Prox1 had inhibitory effect on both cell proliferation and tumor formation. cDNA microarray analysis indicated that the expression of PKCβII was highly suppressed by Prox1 overexpression. In Prox1-expressing Hela cells, mRNA expression of PKCβII was undetectable and protein was significantly down-regulated compared with Mock cells. The selective PKCβII inhibitor, enzastaurin was less effective on cell proliferation of Prox1-expressing cells compared with Mock cells. These results showed that PKCβII was required for tumor cell proliferation. We focused on the regulatory mechanism of PKCβII expression by Prox1. The 5'-promoter analysis of PKC-β gene revealed that the proximal region between 110bp of the promoter containing 2 Sp1 binding sites was responsible for the endogenous promoter activity, and Prox1 did not affect that. Mithramycin A, a relatively specific Sp1 inhibitor, and Sp family knockdown by siRNA on Hela cells decreased PKCβII expression, suggesting that Sp family was important for PKCβII transcription. Interestingly, 5-aza-deoxycytidine (5-Az) treatment restored PKCβII mRNA expression in Prox1-expressing Hela cells, suggesting that 5'-promoter region of PKCβII was methylated in these cells. Actually, bisulfite sequencing showed that the predicted CpG island of PKC-β gene, especially around these Sp1 binding sites, was highly methylated in Prox1-expressing Hela cells compared with Mock cells and 5-Az relieved DNA methylation.

CONCLUSION:
Our study suggested that the decreased PKCβII expression suppressed tumor cell proliferation and Prox1 down-regulated PKCβII expression by enhancing DNA methylation of PKC-β 5'-promoter around Sp1 binding sites. This is the first to report the inverse relationship between Prox1 and PKCβII expression in cancer cells and the novel regulatory mechanism of PKC-β gene expression by Prox1 transcription factor.

Disclosures:
No relevant conflicts of interest to declare.
The Comparison of Fibrin Monomer (FM) Performance to Other Activation Markers (TAT, PF1.2 and DD)
Authors: Patel Priti , Telesca Mayra , Ku De-Hui 
Published: October 2019

Abstract
Abstract 5259
In this study, we attempted to determine the performance of the soluble fibrin monomer complex (SFMC) test in comparison to known activation markers, such as prothrombin fragment 1.2 (PF1.2), thrombin antithrombin complex (TAT) and d-dimer (DD). We have used the STA(R)-Liatest FM and STA(R)-Liatest D-Di kits to measure the SFMC and d-dimer levels.
A total of 25 patients were included in this study. The data and demographic information are shown in the table below.
ID GENDER AGE FM TAT PF1.2 D-dimer 1 M 66 0.42 2.3 1988 0.05 2 F 31 27.93 >60.0 >12000 2.39 3 F 24 3.41 14.1 1367 0.48 4 F 61 2.22 7.7 335 0.63 5 F 18 4.61 6.9 940 0.22 6 F 38 156.52 43.5 >12000 10.37 7 F 20 0.42 2.2 149 0.66 8 F 21 0.42 2.4 143 0.24 9 M 44 >150 15.4 11043 5.5 10 F 42 3.4 <2.0 134 0.24 11 M 46 0 4.8 271 0.3 12 M 76 90.73 29.4 1712 >20.0 13 F 61 3.41 3.8 484 0.52 14 M 1 0.1 3.8 116 0.33 15 F 33 4.2 37 6346 2.03 16 F 35 3.2 2.6 325 0.09 17 F 23 1.1 4.7 232 0.2 18 M 64 2.3 2.6 189 0.15 19 M 1 1.7 2.9 253 0.49 20 M 64 2.2 33.2 451 2.43 21 M 60 1.6 2.9 126 0.42 22 F 55 2.2 2.6 189 0.97 23 F 65 1.3 2.3 79 0.25 24 F 39 3.6 8.1 222 0.18 25 M 2 4.8 <2.0 109 0.53 Reference range >6.0 ug/mL <4 mcg/L 41–372 pmol/L <0.45 ug/mL
The data indicate that in 16 out 25 patients, the SFMC levels are correlated with TAT and PF1.2. Only 3 out of 25 patients have elevated TAT and PF1.2 with normal SFMC levels. Therefore, our data would tend to indicate that SFMC level could be used as an activation marker to assess an ongoing prothrombotic process. In order to more fully determine the efficacy of the SFMC assay, we will obtain a detailed clinical history for the cases shown above.

Disclosures:
No relevant conflicts of interest to declare.
Src Family Kinases Contribute to GI and Gq-Mediated Platelet Activation
Authors: Xiang Binggang , Zhang Guoying , Li Zhenyu 
Published: October 2019

Abstract
Abstract 5263
The Src family kinases (SFKs) play an essential role in collagen- and von Willebrand factor (vWF)-mediated platelet activation. However, the role of SFKs in G protein-coupled receptor (GPCR)-mediated platelet activation is not fully understood, and little is known about the molecular mechanisms by which SFKs are activated by GPCR stimulation. Here we demonstrate that SFKs are activated by the Gq and Gi pathways, respectively and SFKs play important roles in Gq- and Gi-dependent secretion and activation. ADP induced SFK phosphorylation in wild type and Gq−/− platelets, and ADP-induced SFK phosphorylation was inhibited by the P2Y12 antagonist AR-C69931MX or P2Y12 knockout but was not affected by the P2Y1 antagonist MRS-2179. Lyn and Fyn were co-immunoprecipitated with Gi2 in platelets, and ADP-induced SFK phosphorylation was diminished in Lyn−/− platelets. These results demonstrate that ADP induces SFK activation mainly depending on the Gi pathway activated via its receptor P2Y12. Furthermore, epinephrine also dose-dependently induced SFK phosphorylation in mouse platelets. A selective inhibitor of Src family kinase PP2 inhibited ADP-induced Akt phosphorylation, fibrinogen binding, and platelet aggregation. Thus, activation of Gi is sufficient to induce SFK activation, which plays an important role in Gi-dependent platelet activation. We further show that the thrombin receptor PAR4 peptide AYPGKF elicited SFK phosphorylation in P2Y12−/−, but not in Gq−/− platelets, and AYPGKF-induced SFK phosphorylation was inhibited by the calcium chelator dimethyl-BAPTA, suggesting that Gq-dependent SFK phosphorylation is downstream from the Ca2+ signaling. The calcium ionophore, A23187-induced TXA2 synthesis, platelet aggregation and secretion were inhibited by pre-incubation of platelets with PP2. PAR4-induced TXA2 synthesis was also abolished by PP2. Moreover, PAR4-mediated granule secretion, integrin aIIbb3 activation, and aggregation of P2Y12 deficient platelets were partially inhibited by PP2 or a PKC inhibitor Ro-31-8220, but were completely abolished by Ro-31-8220 plus PP2 or dimethyl-BAPTA, suggesting that Ca2+/SFKs and PKC represent two parallel signaling pathways mediating Gq-dependent platelet activation. In summary, SFKs can be activated by Gq/Ca2+-dependent mechanisms and by Gi-dependent mechanisms, and SFKS play important roles in Gq- and Gi-dependent platelet activation.

Disclosures:
No relevant conflicts of interest to declare.
Serum Prohepcidin and Iron Homeostasis in Patients with Breast Cancer
Authors: Grudeva Janet G. , Nenova Ivanka Slavejnova , Terzieva Dora Dimitrova , Spasova Maria Ivanova , Boyadjiev Nikolay Petrov 
Published: October 2019

Abstract
Abstract 5291

Introduction:
Cancer-related anemia (CRA) has multifactorial etiology and complex pathogenesis. It is defined as normochromic, normocytic anemia with reticulocyto-penia and hypoferremia. Hepcidin is recognized as the central factor in causing CRA. Objective: To investigate the changes in the serum levels of prohepcidin (pHp) and markers of iron homeostasis for gathering more data on the pathogenesis of CRA.

Patients and Methods:
The authors analyzed prospectively 46 newly diagnosed women with breast cancer, aged 29–72 years (average 47.5±9.0 SD), who had the same clinical stage, histology and hormonal status. They were diagnosed and treated in the Clinic of Oncology and Hematology - University Hospital for a 2-year period (2009–2010). Serum pHp levels and common markers of iron status including serum iron (sFe), transferrin saturation, soluble tranferrin receptor (sTfR), Zn-protoporphyrin (ZPP), ferritin as well as vitamin B12 and folate were measured before treatment initiation and two months afterwards. Serum pHp was determined by a competitive immunotest. All statistical data was computed by the methods of variational and correlation analyses.

Results and Discussion:
For the two-month follow up interval parameters of full blood count remained without statistically significant deviation. Although the analyzed parameters in the first and second blood samples remained within the reference intervals, the decrease of pHp and the changes of iron-containing substances and ZPP in red blood cells cannot be neglected. However the correlation coefficient (R) between decreased pHp, increased sFe (R=0.314), sTfR (R=0.258), ZPP (R=0.118) and decreased ferritin (R=0.099), were low instead of the expected higher relationship. The obtained results do not support the idea that iron increase and other changes of iron homeostasis in these patients is a direct result of regulatory decrease of pHp. The two-month interval is likely short enough to rule out a potential direct suppressive effect of the specific cytotoxic treatment, and therefore we do not discuss such potential influence on iron homeostasis.

Conclusion:
Iron homeostasis dysregulation is one of CRA components. In its complex pathogenesis several other factors interfere e.g. such related to the disease; therapy related factors; altered erythropoietin production; activation of cytokines (IL-6 is the major promoter of hepcidin production).

Disclosures:
No relevant conflicts of interest to declare.
Impact of Different Immunosuppresive Drugs on the Endothelium. Protective Effect of Defibrotide
Authors: Diaz-Ricart Maribel , Carmona Alba , Palomo Marta , Rovira Montserrat , Escolar Gines , Carreras Enric 
Published: October 2019

Abstract
Abstract 5319
Endothelial dysfunction is a key factor in the development of hematopoietic stem cell transplantation (HSCT) complications due largely to the conditioning treatments. The effect of immunosuppressive therapy on endothelial function has not been explored in detail in this setting. We evaluated the effect of three immunosuppressive drugs: two calcineurin inhibitors (cyclosporine and tacrolimus) and an inhibitor of mTOR (rapamycin). Microvascular endothelial cells (HMEC) were exposed to the three drugs to assess changes in ICAM-1 expression on the cell surface, the reactivity of the extracellular matrix (ECM) generated, and in signaling proteins. Studies were conducted in the absence and presence of defibrotide (DF) to assess its protective effect. Only the exposure of HMEC to cyclosporine (CyA) increased significantly the expression of ICAM-1 (4-fold, p<0.01) and the reactivity of the ECM (surface covered by platelets of 32.1±2.6% vs. 26.5±2.5%, p<0.01). While tacrolimus itself only increased slightly ICAM-1 expression, rapamycin moderately decreased the ECM reactivity. Those changes due to CyA were prevented by DF. CyA induced the activation of AKT and Raf, among others, whose expression was inversely related (4- and 0.5-fold, respectively, with respect to control cells). DF reversed this trend (0.4- and 2.7-fold, respectively, with respect to CyA treated cells). In conclusion, both calcineurin inhibitors showed a proinflammatory effect, though only CyA exhibited significant proinflammatory and prothrombotic effects on the endothelium that correlated with AKT/Raf pathway activation. Interestingly, the protective anti-inflammatory and anti-thrombotic effect of defibrotide seems to occur through the same signaling pathway. In contrast, rapamycin did not exert any significant action on endothelium.

Disclosures:
No relevant conflicts of interest to declare.
Perioperative Heparin Bridging in Patients Receiving Oral Anticoagulation: Meta-Analysis of Bleeding and Thromboembolic Rates
Authors: Yudin Jovana , Siegal Deborah , Lim Wendy , Kaatz Scott , Douketis James , Spyropoulos Alex C 
Published: October 2019

Abstract
Abstract 545

Background:
Periprocedural bridging using unfractionated heparin (UFH) or low molecular weight heparin (LMWH) in patients receiving chronic oral anticoagulation (OAC) is often utilized with the view to reduce the risk of thromboembolic (TE) events. Optimal perioperative anticoagulant methods have not been established. Methods: Systematic review and meta-analysis of published English-language studies from 2001 to 2010 examining bleeding and TE events in patients receiving bridging therapy during temporary OAC interruption for elective procedures. Results: A search of MEDLINE, EMBASE and Cochrane Collaboration databases yielded 32 studies on 6760 bridged patients. Studies were reviewed by 2 independent data collectors (k=0.869). Study quality was generally poor with risk of bias. Thirty-one studies were observational with 1 randomized controlled trial. Low TE risk and/or non-OAC patient groups were used for comparison in 12 observational studies. Major (22/32, 68.8%) and non-major (27/32, 84.4%) procedures were represented. TE events occurred in 67 of 6760 bridged (0.87%; 95% CI 0.40%–1.35%) and 29 of 4897 non-bridged (0.77%; 95% CI 0.24%–1.30%) patients. Using a random effects model, there was no difference in the risk of TE events in bridged versus non-bridged patients (OR 1.02, 95% CI 0.53–1.95). Bridged patients had a significantly increased risk of overall bleeding (OR 5.47, 95% CI 3.89–7.70) and major bleeding (OR 3.43, 95% CI 1.13–10.4) compared to non-bridged patients. There was no difference in TE events (OR 2.44, 95% CI 0.34–17.4) or overall bleeding (OR 2.40 95% CI 0.72–8.05) in patients receiving full versus intermediate/low dose LMWH. Summary: Patients receiving heparin bridging during OAC interruption appear to be at increased risk of bleeding and similar risk of TE events compared to non-bridged patients. Studies of high methodologic quality are needed to develop an optimal anticoagulation strategy and inform clinical decision-making.

Disclosures:
Lim: Leo Pharma: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bayer: Consultancy; Pfizer: Honoraria. Kaatz:Boehringer-Ingelheim: Consultancy, Research Funding, Speakers Bureau; Bristol Myer Squibb: Consultancy, Research Funding; Bayer: Research Funding; National Institute of Health: Research Funding; Canadian Institute of Health Research: Research Funding; Pfizer: Consultancy; Johnson and Johnson: Consultancy; Ortho-McNeil: Consultancy; GlaxoSmithKline: Speakers Bureau; AC Forum: Membership on an entity's Board of Directors or advisory committees; National Certification Board of Anticoagulation Providers: Membership on an entity's Board of Directors or advisory committees; National Blood Clot Alliance: Membership on an entity's Board of Directors or advisory committees.
MARCKS Confers Resistance to Velcade Through Skp2/p27-Mediated Pathway in Multiple Myeloma
Authors: Yang Yijun , Micallef Johann , Chang Hong 
Published: October 2019

Abstract
Abstract 985
Despite advances in clinical care, multiple myeloma (MM) remains an almost universally fatal disease. Velcade, a novel proteasome inhibitor represents a promising new clinical strategy for relapsed and refractory MM. However, only 30–40% of MM patients respond to this treatment. In order to gain insight into the mechanism of velcade resistance, we examined the differentially expressed proteins between 8226S (velcade sensitive) and 8226R5 (velcade resistant) myeloma cell lines using iTRAQ mass spectrometry (MS). The MS based multiple-reaction-monitoring technique (MRM) was then used to independently verify the quantitative differences of the protein expression levels between the two cell lines. We identified several cancer specific proteins to be highly differentially expressed between 8226S and 8226R5 by iTRAQ mass spectrometry. Of particular interest is Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS), a PKC substrate that has been implicated in conferring drug resistance to ovarian carcinoma cells. Differential expression of MARCKS between sensitive and resistant cells was verified by MRM and then confirmed by RT-PCR and Western blot analysis. MARCKS over-expression was also found in additional MM cell lines as well as patient samples that were resistant to velcade. Knockdown of MARCKS through siRNA strategies enhanced velcade sensitivity by increasing apoptosis in the velcade treated resistant cell line 8226R5. Moreover, inhibition of PKC, the upstream activator of MARCKS by enzastaurin significantly blocked phosphorylation of MARCKS leading to a caspase dependent apoptosis in the 8226R5 cells. In addition, enzastaurin enhanced velcade proteosome inhibitory effect and rendered the 8226R5 cells more sensitive to velcade treatment. Combination of velcade with enzastaurin showed a synergistic cytotoxic response in MM cells compared to either drug used alone. Furthermore, knockdown of MARCKS or inhibition of MARCKS phosphorylation by enzastaurin led to a decrease in Skp2 expression and an increase in p27 and p21, as well as a reduction of cyclin E and CDK2, resulting in G1/S cell cycle arrest and an increase in apoptosis in MM cells. Taken together our results suggest that phosphorylated MARCKS plays an important role in velcade resistance in MM and as such may be used as an important therapeutic target. In addition, MARCKS may be a biomarker to select patients that would most benefit from patient specific treatment.

Disclosures:
No relevant conflicts of interest to declare.
Iron Regulatory Proteins in Systemic Iron Metabolism and Erythropoiesis
Authors: Hentze Matthias W. 
Published: October 2019

Abstract
Abstract SCI-22
Imbalances of iron homeostasis account for some of the most common human diseases. Pathologies can result from both iron deficiency or overload. The hepcidin/ferroportin and the IRE/IRP regulatory systems balance systemic and cellular iron metabolism, respectively, and understanding their points of intersection and crosstalk represents a major challenge in iron biology. I will discuss an emerging picture from studies with different mutant mouse lines according to which the “cellular” IRE/IRP system determines “set points” via its targets (including ferroportin and HIF2α). These are then subject to modulation via hepcidin in response to systemic cues.

Disclosures:
No relevant conflicts of interest to declare.