bluuebunny/crossref_metadata_2025_split huggingface dataset:
LPL Is the Strongest Prognostic Factor in a Comparative Study of RNA-Based Markers in Chronic Lymphocytic Leukemia.
Authors: Kaderi Mohd Arifin , Kanduri Meena , Mansouri Mahmoud , Buhl Anne Mette , Sevov Marie , Cahill Nicola , Gunnarsson Rebeqa , Jansson Mattias , Smedby Karin Ekström , Hjalgrim Henrik , Juliusson Gunnar , Jurlander Jesper , Rosenquist Richard 
Published: October 2019

Abstract
Abstract 1254
Poster Board I-276

Introduction
Chronic lymphocytic leukemia (CLL) is a heterogeneous disease with varying clinical outcome, where many patients have an indolent course for many years, whereas others show a more aggressive disease despite treatment. This has prompted the search for biomarkers that can predict outcome in this disease. Recent studies have proposed the RNA expression levels of certain genes, i.e. LPL, CLLU1, TCL1, MCL1 and ZAP70 to be novel predictors of clinical outcome in CLL. However, a comprehensive assessment of these RNA-based markers is still lacking. The current study aimed to investigate the potential of these markers in CLL prognostication, either as single markers or in combination with established markers.

Patients and Methods
By applying real-time quantitative PCR, we measured the RNA expression levels of LPL, CLLU1, TCL1, MCL1 and ZAP70 in 256 newly diagnosed CLL samples from a Scandinavian population-based cohort collected from 1999 to 2002 (median follow-up, 89 months) and correlated with clinical outcome. The expression cut-offs for each RNA marker was determined by constructing ROC curves. Additionally, Binet stage, IGHV mutation status, CD38 expression (cut-off 7%) and the presence of recurrent genomic aberrations (i.e. 11q-, 17p-, 13q- and +12) were evaluated for all cases.

Results
High expression of all RNA-based markers except MCL1 predicted significantly shorter overall survival (OS) and time to treatment (TTT), with LPL being the most significant prognostic marker in both log-rank (Table 1) and Cox univariate regression analyses. In multivariate analysis including the RNA markers, LPL expression was the only independent prognostic factor for OS, whereas both LPL and CLLU1 could predict TTT. When including all established markers, LPL lost its significance in the model, due to its close association to the IGHV mutation status. Once the mutation status was excluded from the analysis, LPL regained its prognostic power in addition to genomic aberrations and CD38. Interestingly, all of the RNA-based markers added further prognostic information to established markers in subgroups of patients, with LPL expression status giving the most significant results. Notably, high LPL expression predicted a worse outcome in favorable prognostic subgroups such as patients with Binet stage A, CD38 negativity or favorable genomic aberrations (Table 2).

Conclusions
Altogether, we conclude that LPL expression appear to be the strongest among the RNA-based markers for prediction of clinical outcome in CLL and thus could potentially be applied in the clinical laboratory to predict outcome, particularly in combination with established markers.

Disclosures
No relevant conflicts of interest to declare.
Neutrophil CD64 Is a Good Indicator of Infection and Sepsis.
Authors: Jou Josep M , Navalón Fulgencio , García Rosario , Brugues Rosa , Nicolás José , Escolar Gines , Diaz-Ricart Maribel 
Published: October 2019

Abstract
Abstract 1347
Poster Board I-369

Introduction
Sepsis is among the top 10 causes of death, but improvements in the diagnostic tests for detecting and monitoring sepsis and infection have been limited in the last years. Neutrophil CD64 expression increases rapidly in the presence of inflammation mediators and in response to infection and tissue damage. We have evaluated changes in the expression of neutrophil CD64 in infected patients in comparison with other markers of infection and sepsis.

Methods
Prospective analysis of 56 blood samples from patients from the intensive care unit at our institution was performed for neutrophil CD64 expression, C-reactive protein (CRP), automated absolute neuthophil count (ANC), and complete manual leucocyte formula including % of bands (BANDS), and % of metamyelocytes and myelocytes (IG). Neutrophil CD64 expression was measured by flow cytometry using a quantitative method (Leuko64TM, Trillium Diagnostics, LLC). Patients were categorized into 5 groups (CLINIC) based on the clinical history and the degree of a systemic inflammatory response, from 1 (no inflammation) to 5 (septic shock). Statistics were performed using linear regression, correlation coefficient, and Passing-Bablock (P-B) regression. Sensitivity (S), specificity (SP), efficiency (E), and positive and negative predictive values (PPV and NPV respectively) were analyzed for all the parameters measured.

Results
Our results showed a correlation with CLINIC of 0.417, 0.552, 0.268, and 0.136 for CD64, CRP, BANDS, and ANC, respectively. P-B regression was only good for CD64, with a slope of 1.03 (0.6-1.4). Percentages (%) of S, SP, E, PPV, and NPV for CD64 were of 81%, 72%, 71%, 46% and 92%, respectively for groups 4 and 5. For CRP, S was of 93% with SP of 20%, E of 38%, PPV of 27%, and NPV of 91%. The remaining parameters showed deficient correlation with CLINIC. Correlations between CD64 and CRP, BANDS, and ANC were of 0.435, 0.342, and 0.01, respectively.

Conclusions
Neutrophil CD64 expression quantitation provides improved diagnostic detection of infection/sepsis compared with the standard diagnostic tests used in current medical practice. CD64 expression showed a better PPV than CRP, and an acceptable NPV. CRP showed deficient SP and E. BANDS, GI, and ANC showed no correlation with CLINIC. CD64 is a new indicator of infection that deserves consideration to be introduced in the daily hematology laboratory analysis.

Disclosures
No relevant conflicts of interest to declare.
Gadd45 Null Mice and Neutrophils/Macrophages Derived From the Bone Marrow of These Mice Exhibit Impaired Innate-Immune/Inflammatory Reponses.
Authors: Salerno Dominic M , Hoffman Barbara , Liebermann Dan A. 
Published: October 2019

Abstract
Abstract 1350
Poster Board I-372
Stress sensor Gadd45 proteins modulate p38-NF-Kb and JNK signaling, which play major roles in leukocyte activation and innate immunity. We have previously documentedthat under conditions of hematological stress, notably acute stimulation with cytokines or inflammation, both gadd45a-deficient and gadd45b-deficient mice exhibited impaired inflammatory responses as indicated by lower percentages of Gr-1-positive cells in the BM and lower numbers of myeloid cells in peritoneal exudates (Gupta et. al Oncogene 25:5539-46, 2006). Recent evidence has implicated Gadd45 proteins in dendritic cell functions that influence effector Th1 responses to inflammation. However, whether gadd45 genes play a role in modulating the myeloid compartment, notably macrophage & granulocyte functions in response to inflammatory stress, remains largely unexplored. To this end, we have employed in vitro & in vivo models of inflammation using BM derived neutrophils and macrophages from WT, gadd45a and gadd45b null mice. The data obtained indicate that chemotaxis and transmigration to various chemo-attractants, including LPS and fMLP, as well as oxidative burst and phagocytic functions were impaired for both neutrophils and macrophages from mice lacking either gadd45a or gadd45b. Furthermore, upon stimulation with LPS, cytokine secretion, notably, but not exclusively IL-12 and TNFa, was significantly reduced in neutrophils and macrophages of gadd45a-/- and gadd455b-/- mice. Western Blot analysis of BM derived neutrophils lacking gadd45a and stimulated with LPS (500ng/mL) exhibited defects in p38 phosphorylation as compared to controls, suggesting a possible mechanism by which the innate response is impaired. P38 phosphorylation in gadd45b null granulocytes stimulated with LPS appeared comparable to what was observed in wt controls. This suggests that gadd45a and gadd45b utilize different signaling pathways to regulate innate-mmune/inflammtory responses. Interestingly, gadd45a, gadd45b & gadd45g null mice injected intraperitoneally with sublethal (25mg/kg body weight) doses of LPS were significantly more susceptible to septic shock compared to wt mice, as indicated by significantly increased morbidity through 5 days post LPS administration. Moreover, 18 hrs. post-injection, the spleens of KO mice were shown to have numerous apoptotic foci in the white pulp, confirmed to be tingible body macrophages ingesting dying cells by IH and IF for macrophage markers. These in vitro and in vivo data suggest a novel role for gadd45 family members in myeloid innate immune responses. Further elucidation of the signaling pathways involved is in progress and is expected to elucidate the molecular basis for the role Gadd45 proteins play in macrophage and granulocyte innate immune functions.

Disclosures
No relevant conflicts of interest to declare.
Nicotinamide Phosphoribosyltransferase (Nampt) Induces NAD+ / SIRT1 Mediated Deacetylation of FOXO3a in Myeloid Cells.
Authors: Lippka Yannick , Thakur Basant Kumar , Skokowa Julia , Welte Karl 
Published: October 2019

Abstract
Abstract 1352
Poster Board I-374
Severe congenital neutropenia (CN) is a heterogeneous disorder of hematopoiesis characterized by a maturation arrest of granulopoiesis at the level of promyelocytes with peripheral blood absolute neutrophil counts below 0.5 × 10/l. G-CSF treatment increases blood neutrophil numbers in more than 90% of individuals with CN. CN is also considered as a pre-leukemic syndrome, since ca. 20% of CN patients develop AML/MDS. Recently we found that NAMPT, a protein involved in biosynthesis of NAD+ was significantly increased in CN patients treated with G-CSF as compared to healthy individuals (Skokowa et al, Nature Medicine, 2009). Increased NAD+ levels correlated with the elevated levels of SIRT1, an enzyme involved in deacetylation of several histone and non -histone proteins (e.g. FOXO3a) by utilising NAD+. In search of the downstream factors regulated by G-CSF/ NAMPT /SIRT1 pathway, we found elevated levels of FOXO3a protein in CN patients treated with G-CSF compared to healthy controls or patients with other types of neutropenia. Therefore, we were interested whether NAMPT/ NAD+ -dependent activated SIRT1 affects FOXO3a levels. Indeed we observed that overexpression of NAMPT leads to the upregulation of FOXO3a protein in 293T cells and endogenous FOXO3a interacts with endogenous SIRT1 in both 293T cells and promyelocytic cell line NB4. The compound FK866 which specifically inhibits NAMPT has recently entered clinical trials as a potential chemotherapeutic agent. As acetylation of FOXO3a is considered to be important for its tumor suppressor function we asked if inhibition of NAMPT using FK866 increases the acetylation of FOXO3a. Indeed we show that acetylation level of FOXO3a is enhanced when 293T and NB4 cells were treated with FK866. Furthermore, increased acetylation of FOXO3a correlates with the decreased interaction between FOXO3a and SIRT1 in 293T cells after treatment with FK866. GADD45, a protein involved in DNA damage repair, is a well known target of FOXO3a. We found that in CD33+ myeloid progenitor cells from G-CSF treated CN patients with high NAMPT / SIRT1 / FOXO3a levels also had low GADD45 mRNA levels. We performed reporter gene assay using luciferase construct containing wild type GADD45 promoter and observed that SIRT1 enhances the FOXO3a mediated downregulation of GADD45 promoter and interestingly NAD+ further augments this effect. Taken together we hypothesized that NAMPT/NAD+ activated SIRT1 mediates deacetylation and activation of FOXO3a protein, leading to downregulation of target genes (e.g. GADD45) with tumor suppression functions which might possibly be involved in the leukemic transformation in CN patients.

Disclosures
No relevant conflicts of interest to declare.
PET Scan Results of NCCTG N0489: Epratuzumab and Rituximab in Combination with Cyclophosphamide, Doxorubicin, Vincristine and Prednisone Chemotherapy (ER-CHOP) in Patients with Previously Untreated Diffuse Large B-Cell Lymphoma.
Authors: Micallef Ivana N. , Maurer Matthew J. , Witzig Thomas E. , Nikcevich Daniel , Kurtin Paul , Cannon Michael W. , Flynn Patrick J. , Soori Gamini S. , Lowe Val , Wiseman Gregory 
Published: October 2019

Abstract
Abstract 137

Background:
FDG-PET/CT after 2–4 cycles of chemotherapy has been shown to be predictive of outcome in patients with aggressive lymphoma. This multicenter NCCTG phase II study was carried out to assess efficacy of ER-CHOP in newly diagnosed DLBCL; functional response as measured by FDG-PET/CT was done after 2 cycles and after 6 cycles of therapy.

Methods:
Patients received immunochemotherapy on the following schedule: epratuzumab 360 mg/m2, rituximab 375 mg/m2, and standard dose CHOP every 3 weeks for 6 cycles. FDG-PET/CT was done after 2 cycles and again after 6 cycles of therapy. All PET scans were centrally reviewed by an expert in nuclear medicine who was blinded to the outcome of the patients.

Results:
107 patients were accrued from Feb 2006 to Aug 2007. 27 patients were ineligible resulting in 80 eligible patients. Baseline patient characteristics for the eligible patients included median age 60 (range 21–82); 56% were male. 80% had advanced stage; 73% had an elevated LDH. Performance score was 0–1 in 71 pts and 2–3 in 9 pts. IPI was 0–1 in 18 pts (23%), 2 in 22 pts (28%), 3 in 29 pts (36%) and 4–5 in 11 pts (14%). PET scans were available on 76 patients (95%); 3 patients completed treatment but scans were unavailable and 1 died prior to cycle 2 PET. The PET CR rate after 2 cycles was 79%. The PET CR rate after 6 cycles was 90%. Using an intent to treat (ITT) analysis, 67 patients (87%) achieved PET CR of which 61 (79%) also completed all 6 cycles of treatment. Based on interim PET after 2 cycles, the EFS at 24 months (EFS24) was 73% for the PET negative patients vs 60% for the PET positive patients (p=0.25); OS at 24 months (OS24) was 83% vs 73% for the PET negative and positive pts, respectively (p=0.17). In comparison, EFS24 for patients with a negative PET after 6 cycles was 80% vs 57% for those with a positive PET (p=0.11); OS24 was 92% vs 57%, respectively (p=0.01). In intent to treat (ITT) analysis, patients that achieved a functional CR had improved overall survival (OS24 = 84%) and event free survival (EFS24 = 74%) compared to patients that failed to achieve a functional CR (OS24 = 50%, EFS24 = 40%, p=0.01 and p=0.006 respectively).

Conclusions:
ER-CHOP every 21 days × 6 cycles results in a functional CR rate of 87%. Early PET scan during therapy does not significantly predict outcome. Achievement of PET negativity by completion of therapy is associated with a good outcome

Disclosures:
No relevant conflicts of interest to declare.
In-Hospital Mortality and Trends Associated with Splenectomy in Patients with Immune-Mediated Thrombocytopenia (ITP).
Authors: Jaglowski Samantha M. , Byrd John C. , Jones Jeffrey A. 
Published: October 2019

Abstract
Abstract 1398
Poster Board I-420

Background:
Splenectomy remains a standard treatment for ITP patients not responding to medical management, but anecdotal reports suggest that use of the procedure is in decline. We studied patterns of use and outcome of splenectomy performed for ITP at the population level.

Methods:
Using data from the Nationwide Inpatient Sample and ICD-9 diagnosis and procedure codes, we identified 39,543 splenectomies among hospital admissions including a diagnosis of ITP (ICD-9 287.3) from 1993-2005. Admissions were characterized by patient and hospital facility characteristics. Laparascopic procedures were identified by published procedure coding algorithms. Factors influencing in-hospital mortality for 2005 were further evaluated using multivariate logistic regression models.

Results:
Annual estimates for incidence of splenectomy are displayed in Figure 1. Between 1993 and 2005, there was a decrease in the total number of splenectomies performed for ITP, with the most significant drop occurring from 1997 to 2000, concurrent with the FDA approval of rituximab. Over the same period, there has been an increase in the proportion of splenectomies performed laparoscopically from 3.4% to 18.6%. Patient gender, age, presence of comorbid malignancy, and Charlson score were not significantly associated with type of splenectomy procedure. Among facility factors, only hospital teaching status was a statistically significant predictor of laparoscopic splenectomy use, early but not later in the observation period. On an annual basis, in-hospital mortality did not vary significantly over the observation period, with risks ranging from 1.5% (95% CI 0.83-2.86%) in 1993 to 4% (95% CI 2.8%-5.7%) in 1997. Annual mortality risk between open and laparoscopic procedures likewise did not significantly differ. However, over the total 13-year observation period there was a >60% increased risk of death with an open versus laparoscopic procedure (OR 1.669, p<0.0001). In 2005, 2869 splenectomy procedures were performed. Multivariate logistic regression models for in-hospital mortality that year found that presence of a malignancy (OR 9.65, p=0.003) significantly increased mortality risk. Charlson comorbidity approached statistical significance (0 v. ≥1, OR 6.83, p=0.087). Hospital bed-size (OR 0.87, p=0.73), location (rural v. urban, OR 3.80, p=0.127), and teaching status (OR 0.39, p=0.203) were not significantly associated with outcome.

Conclusions:
While the overall mortality risk from splenectomy in ITP is low, it is influenced by the presence of malignancy and other comorbid conditions. Further studies designed to evaluate newer medical management strategies (e.g. rituximab, thrombopoeitin mimetics, etc.) versus surgical intervention in these higher-risk populations are warranted.

Disclosures:
No relevant conflicts of interest to declare.
A Multifaceted Strategy to Reduce Inappropriate Use of Fresh Frozen Plasma Transfusions in the Intensive Care Unit.
Authors: Arnold Donald M. , Whittingham Heather , Lauzier Francois , Clarke France , McDonald Ellen , Tkaczyk Andrea , Greiter Angela , Waugh Lily , Crowther Mark A. , Cook Deborah 
Published: October 2019

Abstract
Abstract 1412
Poster Board I-435

Introduction:
The overuse of fresh frozen plasma (FFP) transfusions has been well documented, especially among critically ill patients. In a mixed medical surgical intensive care unit (ICU), we documented that 43% of FFP transfusions were given for indications other than those proposed in published guidelines (Lauzier 2007).

Methods:
We developed a 3-Phase multifaceted behavior-change strategy to curtail inappropriate FFP transfusions, documenting all patients who had FFP, excluding plasmapheresis. Phase I was a 3-month baseline assessment period with no intervention, in which the FFP transfusion orders prescribed at the discretion of the ICU team were recorded. Phase II was a 3-month intervention targeted to all ICU clinicians, comprised of education on the appropriate use of FFP transfusions, audit and feedback of performance indicators, and a pre-order FFP Request Form to specify the indication and the pre-transfusion INR. Phase III was a 9-month assessment period incorporating only the FFP Request Form. At the end of the study, the indications for all transfusions were adjudicated independently in triplicate by 2 ICU clinicians and 1 hematologist, to determine whether each FFP transfusion was a) consistent with published guidelines, b) inconsistent with guidelines but appropriate for the ICU context, or c) inconsistent and inappropriate. Discrepancies were resolved in all cases. FFP orders were not withheld if FFP Request Forms were not completed.

Results:
Chance-corrected agreement (which considers clustered transfusions within patients) between ICU reviewers on whether transfusions were consistent or appropriate versus inconsistent and inappropriate was high (phi = 0.80). During Phase I (3 months), 66 FFP transfusions were administered (n= 26 patients), of which 30 were for bleeding. During Phase II (3 months), 24 transfusions were administered (n = 11 patients), of which 11 were for bleeding. During Phase III (7 months of data), 96 transfusions were given (n= 41 patients), of which 57 were for bleeding. Rates of FFP transfusions per month for all indications and for bleeding indications were 22 and 10, respectfully in Phase I; 8 and 4 in Phase II; and 14 and 8 in Phase III. A FFP Request form accompanied 39 (40.6%) of 96 FFP transfusions in Phase III.

Conclusions:
A multifaceted behavior-change strategy appears to be an effective method of changing inappropriate FFP transfusion practices; however satisfactory uptake of a pre-transfusion FFP Request Form requires consistent reminders. We recommend that transfusion guidelines are improved to explicitly incorporate FFP transfusion criteria appropriate for the ICU setting.

Disclosures:
No relevant conflicts of interest to declare.
Prognostic Significance of Circulating Epstein-Barr Virus (EBV) DNA in Non-Hodgkin's Lymphoma.
Authors: Kang Myoung Joo , Cho Yoo Jin , Ahn Heui June , Kim Shin , Lee Dae Ho , Kim Sang We , Chi Hyun-Sook , Suh Cheolwon 
Published: October 2019

Abstract
Abstract 1936
Poster Board I-959

Introduction:
Epstein-Barr virus (EBV) is well-known as an etiologic agent of lymphoproliferative disorders. Almost all types of sporadic non-Hodgkin's lymphoma (NHL) of both B-cell and T-cell origins have been demonstrated to have EBV-infected tumor cells. However, the prognostic significance of EBV viral load has not been established yet. Methods: To monitor quantitative EBV DNA load, real time quantitative PCR of EBV BZLF1 DNA was done using RealArt® EBV LC PCR reagents (Artus) from whole blood samples drawn from newly diagnosed NHL patients at the time of diagnosis and during the follow-up. Results: Between July 2006 and March 2009, a total of 233 patients (including 120 [51.5%] diffuse large B-cell lymphoma [DLBCL]; 18 [7.7%] natural killer (NK)/T-cell lymphoma; and 11 [4.7%] peripheral T-cell lymphoma, unspecified) were reviewed and analyzed. Among them, 33 (14%) patients, had a detectable EBV DNA (positive group) while 200 (85%) patients did not (negative group). Those in positive group had a more advanced disease and a higher LDH level than those in negative group (P=0.003 and 0.007, respectively). With a median follow up of 11.3 months (range 1.2-34.9), 14 (42.4%) of the 33 patients in positive group had a progressive or relapsed disease while 30 (15%) of the 200 patients in negative group did (P<0.001). One-year overall survival (OS) rate was 72.5% in positive group while that was 82.9% in negative group (log-rank P=0.001). Interestingly, conversion to negative EBV DNA during the follow-up was associated with a lower probability of progressive or relapsed disease (P=0.004). NK/T-cell lymphoma patients having an EBV DNA survived shorter than those having not. However, in DLBCL patients, there was no difference in survival between those having an EBV DNA and those not. Conclusions: The presence of EBV DNA at diagnosis and the negative conversion of EBV DNA during the follow-up might have prognostic significance in patients with NHL.

Disclosures:
No relevant conflicts of interest to declare.