bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of Homeostatic Reticulocyte Aging between Normal and Belgrade Rats Using Flow Cytometry.
Authors: Wiczling Pawel , Krzyzanski Wojciech 
Published: April 2020

Abstract
The purpose of this work was to obtain and compare reticulocyte age distribution of normal and Belgrade (b) rats. The Belgrade rat has a microcytic, hypochromic anemia associated with abnormal reticulocyte iron uptake. Samples of blood were drawn from two homozygous rats (b/b) and three heterozygous rats (t/b). Cells were cultured over 12 hours and sampled at 3 hour intervals. The reticulocyte count was performed using a staining solution composed of thiazole orange and LDS 751, a DNA binding agent. LDS 751 was used to separate RBC form nucleated cells. The differentiation between mature RBC and reticulocytes was done by means of presence of residual RNA, which also served as a marker of reticulocyte age. The in vitro changes of the fluorescence signal of the reticulocyte population allowed us to obtain the unique relationship between the signal and the age for an individual cell based on the previously developed structured population model of cultured reticulocytes. This relation was further used to transform the in vivo signal distribution into the age distribution. The obtained reticulocyte age density was used to characterize age dependence of reticulocyte appearance in blood and death. The Belgrade rats have reticulocytosis of 13.6 ± 4% whereas control rats have reticulocytes in normal range of 2.15 ± 0.4%. The aging of reticulocyte was characterized by a relative time, RT, that a reticulocyte needs to become a mature RBC. For the normal reticulocytes the RT that an average reticulocyte spent in blood is 0.84 ± 0.02 days and ranges from 0 to 2.0 days. For Belgrade rats the time an average reticulocytes in blood needs to become a mature RBC is 1.18 ± 0.08 days and ranges from zero 0 to 2.2 days. In conclusion, the Belgrade reticulocytes are younger by about 0.4 days. The RT distributions for Belgrade rats are wider than for normal rats. Additionally, the increased death of Belgrade reticulocytes in circulation is observed. In this work the new method of studying aging of cells was successfully applied for anemic rats. It allowed us to characterize in vivo aging of normal and Belgrade reticulocytes.
Role of Gamma-Carboxyglutamic Acid (GLA) Domain in Phosphatidylserine (PS)-Regulated Activity of Factor IXa.
Authors: Majumder Rinku , Craft Judson , Rezaie Alireza , Barry R. , Monroe Dougald 
Published: April 2020

Abstract
Both factors Xa and IXa are vitamin-K-dependent serine proteases that consist of disulfide-linked heavy and light chains. The heavy chain contains the serine protease active site, while the light chain contains the GLA (gamma-carboxyglutamic acid) domain, EGF-I (epidermal growth factor-like region) and EGF-II domains. Effect of PS on FXa: We have extensively studied the interaction of factor Xa with 1, 2-dicaproyl-sn-glycero-3-phospho-L-serine (C6PS). C6PS is a soluble analogue of phosphatidylserine, which is present in platelet membranes; it serves as a model for the effect of membrane-bound PS on factor Xa activity and structure. We located three C6PS binding sites to different domains of factor Xa using a combination of activity, circular dichroism, fluorescence, and equilibrium dialysis measurements, showing that :
the Gla domain binds C6PS only in the absence of calcium (kd ∼ 1 mM); a calcium-dependent, regulatory, PS-binding site exists in the EGFN domain when linked by calcium to the Gla domain; and a second calcium-requiring site exists in the EGFC-catalytic domains and shares residues with the substrate recognition site.
We have shown that full functional response to C6PS requires linkage of the Gla, EGFNC, and catalytic domains in the presence of calcium. Recently we have observed that the proteolytic activity of des EGFN factor Xa is not affected in the presence of C6PS, locating more precisely a significant component of the PS-triggered regulatory site to the EGFN domain. Efect of PS on FIXa: We have previously reported that C6PS induces a calcium-dependent conformational change in factor IXa that regulates the amidolytic and proteolytic activities. We have also shown that factor IXa binds 2 molecules of C6PS. Here we have examined the role of the GLA domain in this C6PS regulation of factor IXa using a GLA-domainless variant of factor IXa (GD-IXa). In the absence of the GLA domain, binding to C6PS has no effect on the rate of proteolytic or amidolytic activity of GD-IXa toward both factor X and synthetic substrates. The binding of C6PS to GD-IXa studied using intrinsic tryptophan fluorescence (Kd = ∼50μM) was different from that for native factor IXa (Kd = ∼2μM). The critical micelle concentration (CMC) of C6PS under the conditions of these experiments (∼300μM) was much greater than the C6PS concentrations used in our experiments. We conclude that:
C6PS does bind to GD-IXa, but at a reduced affinity compared to factor IXa; C6PS does not regulate the amidolytic or proteolytic activities of factor IXa in the absence of GLA domain.
This could be due to two C6PS sites existing in FIXa, with the regulatory site requiring the Gla domain (as observed for FXa), or to a requirement for the Gla domain for the regulatory activity of a single site. Future studies looking at the stoichiometry of binding of C6PS with GD-IXa will help distinguish between these possibilities.
Supported by Supported by NHLBI (HL 072827).
Overall Survival in Erythrodermic Cutaneous T-Cell Lymphoma: An Analysis of Prognostic Factors in a Cohort of Erythrodermic Cutaneous T-Cell Lymphoma Patients.
Authors: Vidulich Kelley A. , Talpur Rakhshandra , Bassett Roland L. , Duvic Madeleine 
Published: April 2020

Abstract
Most common cutaneous T-cell lymphomas (CTCL) are Mycosis fungoides (MF) and a leukemic, erythrodermic (E) variant known as Sézary Syndrome (SS). Blood (B) criteria were not previously considered for staging purposes and are evolving. E-CTCL can be divided into E-MF versus SS based on new criteria, B0-B2, published by the ISCL guidelines. Historically, overall survival (OS) of “SS” patients is 2–4 years. To determine if hematologic staging and prognostic variables affects overall survival, we retrospectively studied 1197 MF/CTCL patients seen at MDACC since 1987 from which were identified 124 (10%) patients with E-CTCL. Median age at diagnosis was 63 (range 0–89), 71 males and 53 females. Hematologic (H) stage was based on quantitative Sézary cell counts (manual or by flow as absolute CD4+26−). Overall survival curves were estimated by Kaplan and Meier and compared using log-rank tests. Median OS all 124 E-CTCL patients was 5.1 years [range=0.4–18.6]. We also considered patients divided by the H0–H4 staging system (Russell-Jones 2000). For H0–H2, OS was 7.6 years, H3 was 5.4 years, and H4 was 2.4 years (p=0.011). Treatment with systemic steroids, advanced age, increased serum LDH at presentation, WBC &gt; 20,000 were significant prognostic factors. Large cell transformation (p=0.758), positive T-cell receptor gene rearrangement in the skin (p=0.54), stage of disease (p=0.955), prior exposure to multiple treatments (p=0.953), and CD4:CD8 ratio (p=0.068) were not significant. In conclusion, we provide evidence that absolute Sézary cell counts can be used to define three groups of patients with E-CTCL with different overall survival patterns and propose a modification to the ISCL B ratings as follows: B0, absolute Sézary cell count &lt; 1.0 KuL−1; B1: absolute Sézary cell count ≥ 1.0–10.0 KuL−1; and B2: absolute Sézary cell count &gt;10.0 KuL−1. Serum LDH and CD4/8 ratio may be used to estimate tumor burden, as they correlate with Sézary counts.
Race and Non-Hodgkin’s Lymphoma: Adverse Impact of Race and Treatment Delays on Survival. A SEER-Medicare Population Study (1995–2003).
Authors: Vance Katisha T. , Kilgore Meredith L. , Yun Huifeng , Gary Lisa C. , Foran James M. 
Published: April 2020

Abstract
Background: Despite a lower incidence of non-Hodgkin’s lymphoma (NHL) and specifically diffuse large B-cell lymphoma (DLBCL), the most common histologic subtype, previous studies suggest that blacks have a higher disease-specific mortality from NHL than whites. To study this disparity, we looked at the impact of race, treatment delays and the use of rituximab on survival in NHL and DLBCL. To control for access to care, we focused on a population of patients with Medicare benefits.
Methods: After establishing a SEER-Medicare Data-Use Agreement and obtaining IRB approval, we used the linked SEER-Medicare database for NHL patients diagnosed from 1995 through 2002, with claims data through 2003. Multivariable logistic regressions were used to assess the difference between race and rates of survival, NHL histology, and the likelihood of receiving treatment within 90 days and of receiving rituximab at any time during the course of NHL treatment.
Results: Among 41,704 NHL cases, 35,014 (85%) cases were in whites and 2,287 (5.5%) were in blacks. Of the 14,831 total cases of DLBCL, 12,449 (84%) were in whites and 715 (4.8%) were in blacks. Despite a lower incidence of NHL, blacks had a lower 1-year survival for NHL compared to whites (OR 0.68 [95% CI 0.61–0.76]). Blacks also had a lower 1-year survival for DLBCL compared to whites (OR 0.63[95% CI 0.53–0.75]). Patients treated within 90 days of diagnosis had improved 1-year survival for NHL (OR 1.10 [95% CI 1.03–1.18]) and for DLBCL (OR 1.60 [95% CI 1.47–1.75]). When controlling for age, gender and income, blacks were less likely than whites to be treated within 90 days of a NHL diagnosis (OR 0.599 [95% CI 0.526–0.681]). The use of rituximab was associated with improved 1-year survival for NHL (OR 3.67 [95% CI 3.37–4.00]) and specifically for DLBCL (OR 3.01 [95% CI 2.66–3.40]). However, blacks were significantly less likely to receive rituximab for NHL (OR 0.55 [95% CI [0.47–0.63]) and specifically for DLBCL (OR 0.71[95% CI 0.57–0.90]). An analysis of two and three year survival for NHL and DLBCL yielded similar results for the impact of race, treatment delays and use of rituximab.
Conclusions: Blacks are diagnosed with NHL and specifically DLBCL less frequently than whites but have significantly lower 1-year survival rates. Blacks were significantly less likely to begin treatment within 90 days of a NHL diagnosis. In addition, blacks were significantly less likely to receive rituximab despite presumed equal access to healthcare with Medicare benefits. The reason for this racial disparity is unclear. Studies to further characterize the disparity are needed.
Genomic Profiling of Different Subtypes of B-Cell Non-Hodgkin’s Lymphoma Using High-Density Single Nucleotide Polymorphism (SNP) Microarrays.
Authors: Kato Motohiro , Nakazaki Kumi , Takeuchi Kengo , Yamamoto Go , Nannya Yasuhito , Sanada Masashi , Muto Satsuki , Chiba Shigeru , Mori Shigeo , Kobayashi Yukio , Kurokawa Mineo , Ogawa Seishi 
Published: April 2020

Abstract
B-cell non-Hodgkin’s lymphoma (B-NHL) is a constellation of mature B-cell neoplasms characterized by distinct pathological and molecular genetic features, including diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), MATL-type lymphoma (MALT-L). These B-NHLs undergo a wide variety of genetic alterations, including gains and losses of genetic materials, as well as abnormalities in allelic composition, and these alterations may collectively comprise unique genomic profiles specific to different subtypes of B-NHL, which are tightly linked to the pathogenesis of each B-NHL subtype. Recently, we have developed a robust method (CNAG/AsCNAR) to detect allelic imbalances and other copy number changes in cancer genomes using SNP-genotyping microarrays (Nannya et al. Cancer Res, 2005 and Yamamoto et al. Am J Hum Genet, 2007). Without depending on the availability of constitutive genomic DNA, it enables sensitive detection of allelic imbalances including loss of heterozygosity (LOH) as well as copy number (CN) alterations in the face of 70–80% of normal cell components. In the current study, we performed SNP-chip/CNAG/AsCNAR analysis of 171 B-NHL specimens, including 65 cases of DLBCL, 61 cases of FL, and 45 cases of MALT-L in order to comparatively investigate the unique genomic profiles of different B-NHL subtypes. A large number of gains and losses of chromosomal/genomic segments as well as allelic imbalances were identified. While individual genomic profiles are substantially variable among different cases, they collectively showed a characteristic genomic profile in each B-NHL subtypes; +1q, +2p and +18 are common to DLBCL and FL, while +3 and +11q were more frequently found in DLBCL and +11p was more characteristic to FL. MALT-L also showed +3 and +18, but rarely had +1q and +2p. CN neutral LOH due to mitotic recombination (uniparental disomy; UPD) events are frequently found in DLBCL (70%) and FL (54%), but less common in MALT-L. UPD most commonly involves 1p, 1q, 6p, 9p and 12q. In total, 29 loci of high-grade amplification were identified. Among these recurring amplifications were observed at 1q and 2p, which involves FCGR2B, and cRel genes, respectively. These amplifications were detected in both DLBCL and FL, but rare in MALT-L. Total 14 loci of homozygous deletion were also detected and differentially distributed among different B-NHL subtypes. In conclusion, SNP-chip with CNAG/AsCNAR analysis revealed characteristic genomic signatures of distinctive B-NHL subtypes, implicating their unique pathogenesis.
Red Blood Cell Derived Microparticles Are Thrombogenic in Mouse Models of Atherosclerosis.
Authors: Isobe Hirotaka , Perkmann Thomas , Breuss Johannes M. , Binder Christoph J. , Boulanger Chantal M. , Binder Bernd R. 
Published: April 2020

Abstract
In normal individuals microparticles (MPs) derived from leukocytes, erythrocytes, platelets and endothelial cells are found in the circulation. When the thrombogenicity of microparticles was measured in vitro by their capacity to generate thrombin (thrombin generating activity, TGA) in normal control plasma we found the highest in vitro activity to be generated by red blood cell derived MPs and the lowest by monocyte derived MPs. While all MPs exhibited Annexin V binding, indicating the presence of negatively charged phospholipids on the surface, only monocyte and smooth muscle cell (SMC) derived MPs expressed tissue factor and their TGA was dependent on the “extrinsic” factor VII dependent pathway. All other MPs utilized the “intrinsic” factor XI, factor IX and VIII dependent pathway. In vivo, however, only MPs activating the “extrinsic” pathway were active as shown by TAT formation upon injection of MPs in wt C57/bl6 mice (3 minutes after injection). When, however, not directly thrombogenic MPs derived from red blood cells were co-injected with tissue factor bearing MPs, TAT formation was significantly enhanced, indicating that non-directly thrombogenic MPs need tissue factor “priming” to become thrombogenic. To prove this hypothesis, we injected red blood cell (RBC) derived MPs (106–108) into wild type C57BL/6 mice and into ApoE−/− or LDLR−/ − deficient mice, fed a high lipid diet. In wild type C57BL6 mice red blood cell derived MPs did not significantly increase TAT complexes as compared to PBS (RBC derived MPs 108 6.2±0.8 vs. PBS 4.1±1.3 mg/L). When, however, such RBC derived MPs were injected into LDLR−/− mice fed a high lipid diet, TAT complexes significantly increased (PBS 4.4±0.5 vs RBC derived MPs 108 15.8±1.9mg/L). Similar results were obtained also in ApoE−/− mice. This indicates that tissue factor “priming” also occurs in atherosclerotic mice, likely by tissue factor expressed on the plaque or by plaque derived tissue factor bearing MPs. To determine whether local tissue factor expression causes also local fibrin formation upon injection of red blood cell derived MPs, the aortic arch of such mice were analyzed for local fibrin deposition after injection of red blood cell derived MPs. In fact red blood cell derived MPs did not only induce higher TAT complexes in LDLR−/− mice but also increased the number of fibrin positive plaques as compared to PBS. This indicates that red blood cell derived MPs are thrombogenic in animal models of atherosclerosis. When MPs isolated form the arterial blood of atherosclerotic patients undergoing carotid endarterectomy were analyzed, it was found that MPs from symptomatic patients were more thrombogenic and that this increased thrombogenicity was due to increased number of red blood cell derived MPs. This indicates that red blood cell derived MPs might also cause thrombus formation in atherosclerotic patients.
Predictors of Diagnosis Delay and Complications for Newly-Diagnosed Myeloma Patients.
Authors: Friese Christopher R. , Abel Gregory A. , Magazu Lysa A. , Neville Bridget A. , Richardson Lisa C. , Earle Craig C. 
Published: April 2020

Abstract
Background: The increasing incidence in multiple myeloma (MM), coupled with improved efficacy of therapies, warrant population-based studies of diagnosis delay which could lead to disease-related complications.
Methods: We analyzed tumor registry data and provider claims in the SEER-Medicare database, a geographically and racially diverse sample of Medicare enrollees with confirmed MM. Using extant selection criteria to study eligible MM patients, we identified claims with anemia (An) or back pain (BP) codes, conditions related to MM. We estimated the period of time between first claim for An or BP and confirmed MM diagnosis. We defined delay as ≥98 days (the median) between initial An or BP claim and MM diagnosis, and estimated logit models to predict sociodemographic and clinical factors associated with delay. We also estimated logit models to predict the likelihood of renal or skeletal complications following MM diagnosis. The study period was one year before, through six months following, diagnosis. Analyses were adjusted for a known 30-day lag time in MM diagnosis dates in SEER.
Results: From the analytic sample of 5,185 patients, 3,600 had a diagnosis for An (2,576) and/or BP (2,012) prior to MM diagnosis. Male gender (OR 0.73, 95% CI 0.64–0.84) and northeast region (0.79, 0.65–0.96) decreased the likelihood of delay. Over 60% of patients had ≥ 1 Charlson comorbidity; higher Charlson scores and the presence of both An and BP increased likelihood of delay, but were strongly correlated, and an interaction term with these variables in the model was significant. Of the 5,112 patients with no history of renal or skeletal complications, 1,742 (34%) experienced a complication within six months after MM diagnosis. After adjustment for gender, age, race, and subsequent chemotherapy use, patients with: higher Charlson scores (1.17, 1.03–1.33); initial diagnosis during an inpatient stay (2.40, 1.92–3.00), and; no UPEP, SPEP, or bone marrow biopsy as part of the MM workup (1.68, 1.48–1.92) were more likely to experience a complication (29% of patients had neither PEP nor biopsies recorded). Delay between An or BP diagnosis and MM was not associated with likelihood of complication (0.96, 0.91–1.01).
Conclusions: Despite the presence of two classic signs and symptoms of MM, gender and geography significantly influence diagnosis delays. The surprising finding that both anemia and back pain delay diagnosis is partly explained by increased comorbidity in these patients. These findings highlight the importance of considering MM as part of the routine workup for anemia in elderly patients. The existence of comorbidity in these patients may obscure the clinical presentation, resulting in delayed diagnosis. Underuse of common tests to diagnose myeloma may result in poor outcomes, and could be a focus of quality improvement efforts.
Annexin 2 Mediates Plasminogen-Related Recruitment of Neovascular Mural Cells in Lymphoma Angiogenesis.
Authors: Hajjar Katherine A. , Ruan Jia , Sui Guangzhi , Deora Arunkumar B. , Church Sarah , Cohen-Gould Leona , Rafii Shahin , Lyden David C. , Ling Qi 
Published: April 2020

Abstract
Malignant tumor progression requires the development of tumor-associated blood vessels through co-option of nearby host vascular cells, recruitment of marrow-derived progenitor cells, or both. The annexin 2 (A2)/p11 heterotetramer is a cell surface co-receptor for the fibrinolytic proteins plasminogen (Plg) and tPA, and strongly augments the catalytic efficiency of vascular cell plasmin generation. We showed previously that A2−/ − mice display impaired postnatal angiogenesis in the corneal pocket, oxygen-induced retinopathy, and Matrigel plug assays. Here, we examined neovascularization of experimental lymphoma in mice with fibrinolytic deficiency states. For both EL4, a T cell lymphoma, and B6RV2, a B cell lymphoma, growth of implanted tumor cells was rapid in wildtype and tPA−/ − mice, but nearly flat in both A2−/ − and Plg−/ − mice. Immunohistochemical staining of EL4 tissue on day 8 revealed reduced vascular density, frequent intravascular fibrin thrombi, and dilated microvessels in A2−/ −, compared with A2+/+, mice. Electron microscopy and immunofluorescence revealed a striking paucity of pericytes and secondary dropout of endothelial cells within A2−/ − tumor microvessels. Flow cytometric analysis of circulating progenitor cells showed reduced recruitment of VEGFR1+/CD11b+ hematopoietic precursors from bone marrow to blood in A2−/ − versus A2+/+ tumor-bearing mice. In lethally irradiated A2−/ − mice, moreover, tumor growth was rescued completely upon engraftment with A2+/+ marrow. Transplantation of green fluorescent protein (GFP)-labeled A2+/+ bone marrow revealed the appearance of abundant marrow-derived CD11b+/CD68+ cells closely apposed to developing tumor neovessels. Many of these cells also express the pericyte marker, NG2. Transplantation of A2+/+ marrow, finally, restored the investment of tumor microvessels with α-smooth muscle actin-positive pericytes. These data suggest that A2 contributes critically to tumor angiogenesis in experimental lymphoma in mice, by enabling recruitment of A2+ myelomonocytic cells that promote neovascular stabilization by pericyte-like cells. The data suggest further that targeting the A2/p11 system could offer a novel strategy for the treatment of malignant lymphoma.