bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hematopoietic Progenitor Cell Counts Is a Useful Indicator To Determine Appropriate Time For Peripheral Blood Stem Cell Collection
Authors: Kong Sun-Young , Shim Hyoeun , Lee Se-Na , Kong Jung-Hee , Eom Hyeon-Seok , Park Byung-Kiu , Park Hyeon Jin , Yun Tak , Lee Hyewon , Song Yoon Kyung 
Published: October 2019

Abstract

Background
The optimal peripheral blood stem cell (PBSC) collection is a key step for successful outcome in hematopoietic stem cell transplantation (HSCT). Many indicators including preharvest white blood cell (WBC), mononuclear cell (MNC), and CD34 positive cell counts have been used for deciding the adequate time for collection of PBSCs, but each indicator has limitations. Here we investigated hematopoietic progenitor cell (HPC) count as an indicator for PBSC collection. Methods: Data from 851 autologous PBSC collections from 233 patients at the National Cancer Center, Korea, were analyzed. The correlations between harvested CD34 cell counts with preharvest WBC, MNC, CD34 cell counts, and HPC were analyzed, as were correlations by disease and mobilizing agent. Also how the outcome for engraftment can be predicted based on HPC count was studied. Results: The median age of patients was 41 years (range 0.1-72 years). The most frequent diseases were multiple myeloma (n=64) and non-Hodgkin lymphoma (n=56). The correlation coefficient between collected CD34 cells and preharvest CD34 count was (r=0.669, p<0.001), followed by preharvest HPC count (r=0.419, p<0.001), preharvest MNC (r=0.190, p<0.001) and preharvest WBC (r=0.014, p=0.679). The most adequate cut-off value for obtaining >1x106 CD34+ cells/kg at first time of PBSC was 24.0 HPCs/μL with sensitivity and specificity of 67.7% and 74.3% respectively. The cutoff as 28.0 HPCs/μL was adequate for obtaining 2.0 x106 CD34+ cells/kg with sensitivity and specificity of 73.7% and 72.2% respectively. HPC was well correlated with CD34 in PBSC of patients with multiple myeloma (r=0.326, p=0.009), non-Hodgkin lymphoma (r=0.353, p=0.008), especially diffuse large B-cell lymphoma (r=0.810, p<0.001) and acute leukemia (r=0.998, p<0.001). HPC was a better indicator for non-cyclophosphamide (r=0.337, p<0.001) than cyclophosphamide-based chemomobilization (r=0.572, p=0.052). Infused number of HPCs did not affect the times to engraftment of platelets (p=0.896) and neutrophils (p=0.953), though CD34 count of infusion had positive effect on platelet engraftment (p=0.017). Conclusion: HPC count represented good correlation with CD34+ and high area under the curve. Considering advantages of ease for use and cost-effectiveness than those of CD34 count, HPC is a good surrogate marker to determine appropriate timing for PBSC.

Disclosures:
No relevant conflicts of interest to declare.
Post-Transplant Cyclophosphamide Following a Myeloablative Conditioning For Hematologic Malignancies Receiving An Unmanipulated Haploidentical Bone Marrow Transplant
Authors: Raiola Anna Maria , Ghiso Anna , Dominietto Alida , Di Grazia Carmen , Gualandi Francesca , Varaldo Riccardo , Galaverna Federica , Van Lint Maria Teresa , Lamparelli Teresa , Bregante Stefania , Miglino Maurizio , Ballerini Filippo , Bacigalupo Andrea 
Published: October 2019

Abstract

Background
Unmanipulated family haploidentical transplants are becoming increasingly used in patients lacking an HLA identical family donor, or a well matched unrelated donor.

Aim of the study
This is an updated analysis of our program of haploidentical bone marrow transplant (hBMT) , followed by post-transplant high dose cyclophosphamide (PT-CY) in 95 patients with hematoloigic malignancies receiving a myeloablative conditioning.

Methods
The conditioning regimen consisted of thiotepa, busulfan, fludarabine (TBF) (n=47) , thiotepa melphalan fludarabine (n=5) , or 9.9-12 Gy TBI + fludarabine (n=43). The patients’ median age was 49 years (17-74); the disease status at the time of transplant, was first complete remission (CR1, n=29) , CR2 (n=23), or active disease (n=38,40%). The diagnosis was AML (35%), ALL (24%), MDS (12%) myeloproliferative disorders (12%), other malignancies (17%). The total nucleated cell dose infused was 3.37 (1.4-9.17). The median follow up for surviving patients is 440 days (90-930). Graft versus host disease (GvHD) prophylaxis consisted in PT-CY on day+3 and +5 , cyclosporine (from day 0) , and mycophenolate (from day +1).

Results
The cumulative incidence of neutrophil (PMN) engraftment was (90%) and all surviving patients on day +30 had full donor chimerism. The median day for neutrophil engraftment was day +17 (11-32). The cumulative incidence (CI) of grade II-III acute GvHD was 14%, and of moderate chronic GvHD 8%, severe cGvHD 9%. The CI of transplant related mortality (TRM) is respectively 7%, 17%, 26% for patients in CR1, CR2, or with active disease. The CI of relapse is respectively 17%, 30%, 37% for patients in CR1, CR2, or with active disease. The overall actuarial survival is respectively 75%, 44%, 32% for patients in CR1, CR2, or with active disease (p=0.002) (Fig.1). The most frequent cause of death was leukemia (26%) followed by infections (9%).

Conclusions
These data confirm our first report on 50 patients and support the use of myeloablative conditioning regimens, followed by h-BMT with PT-CY. The risk of acute and chronic GvHD seems relatively low , as well as the overall transplant related mortality. We have not seen the excess relapse rate reported when patients are given a non myeloablative regimen, and disease control has been satisfactorily both for patients in remission, as well as for patients with active disease at the time of transplants.

Disclosures:
No relevant conflicts of interest to declare.
Hypothyroidism and Cardiac Complications In Thalassemia Major Patients
Authors: Gamberini Maria Rita , Meloni Antonella , Rossi Giuseppe , Secchi Giuseppina , D'Ambrosio Alfonso , Macchi Silvia , Pulini Stefano , De Franceschi Lucia , Vallone Antonino , Lombardi Massimo , Pepe Alessia 
Published: October 2019

Abstract

Introduction
In the non-thalassemic population hypothyroidism has been associated with an increased risk of cardiac disease while the link thyroid-heart disease has been little explored in thalassemia major (TM). This retrospective cohort study aimed to systematically evaluate in a large historical cohort of TM in the cardiovascular magnetic resonance (CMR) era whether hypothyroidism was associated with a higher risk of heart complications (heart failure, arrhythmias and pulmonary hypertension).

Methods
From a cohort of 957 TM patients who underwent CMR for myocardial iron overload (MIO) assessment, quantification of biventricular function and detection of myocardial fibrosis within the MIOT network (Myocardial Iron Overload in Thalassemia), we identified 115 (12%) hypothyroid patients. Each hypothyroid patient was matched by sex and age (at the time of the CMR) with two non-hypothyroid patients, creating 115 triples. A cardiac event was considered valid if diagnosed at an age older than the hypothyroidism’s onset age for the hypothyroid patient in the belonging triple.

Results
Hypothyroid and non-hypothyroid patients had comparable MIO, but hypothyroid patients showed significantly lower biventricular stroke volume index, ejection fraction and left ventricular cardiac index. Accordingly, the prevalence of overall heart dysfunction (LV, RV or both) was higher in hypothyroid patients (43.5% vs 33.5%, P=0.0314). Hypothyroid patients had a significant higher frequency of heart failure (19.1% vs 9.1%, P=0.003) and arrhythmias (11.3% vs 4.3%; P=0.003).
Figure1 shows odds ratios (OR) estimating the relationship between hypothyroidism and cardiac involvement. Hypothyroid patients had a significant higher risk of heart dysfunction, heart failure and arrhythmias, also adjusting for the endocrine co-morbidity.

Conclusions
Hypothyroidism seems to increase the risk for heart failure, arrhythmias and heart dysfunction in TM patients. Our data confirm the link thyroid-heart disease also in TM patients and they stress the need to prevent hypothyroidism in this population.

Disclosures:
No relevant conflicts of interest to declare.
Spi-C Negatively Regulates Murine Eosinophil Differentiation
Authors: Schollaert Kaila L , Stephens Michael R , Mingler Melissa K. , Rothenberg Marc E. , Fulkerson Patricia C. 
Published: October 2019

Abstract
Eosinophils are bone marrow-derived granulocytes that normally comprise less than 5% of leukocytes in the blood, but can be found in higher numbers in tissues such as the bone marrow and gastrointestinal tract. Eosinophilia is associated with numerous clinical disorders including atopic diseases, parasitic infections, hypereosinophilic syndrome, and cancer. Expression of Spi-C was originally reported to be limited to B cell and macrophage lineages, but we have reported that upregulation of Spi-C expression within the lung was dependent on the presence of eosinophils in an experimental chronic asthma model. In this study we investigated the role of Spi-C during eosinophil differentiation. At baseline, we observed increased numbers of bone marrow and splenic eosinophils in Spi-C-deficient (Spi-CKO) mice. Stimulation of low-density bone marrow (LDBM) cells with the eosinophil-promoting cytokine IL-5 resulted in 100-fold increase in Spi-C mRNA expression. Cultured Spi-CKO LDBM cells had an accelerated rate of eosinophil progenitor (EoP) differentiation which yielded higher numbers of eosinophils with increased effector functions, including enhanced chemotaxis, granule protein production and release compared to wild-type mice. In addition, induction of asthma resulted in amplified airway eosinophilia in Spi-CKO mice compared to wild-type controls. Together our data indicate that Spi-C negatively regulates eosinophil differentiation during homeostasis and disease. Defining the molecular regulators of eosinophil differentiation and function will undoubtedly provide key information with clinical applications.

Disclosures:
Rothenberg: Teva Pharmaceutical: Consultancy, Ownership Interest receiving Other; Immune Pharmaceutical: Consultancy, Membership on an entity’s Board of Directors or advisory committees, Ownership Interest receiving, Ownership Interest receiving Other.
Platelet Glycoprotein VI Haplotypes and The Generation Of Microparticles In Health and Disease
Authors: Ware Jerry , Dent Judith 
Published: October 2019

Abstract
Platelets are well known for their role in hemostasis, thrombosis, and wound repair. However, there is growing evidence for established platelet paradigms being relevant to other biological processes, such as inflammation and cancer. As one example, the platelet-specific collagen receptor, glycoprotein (GP) VI, has been shown to exacerbate rheumatoid arthritis in mouse models of the disease and also support experimental metastasis. GPVI exists as a membrane surface receptor that once engaged is capable of eliciting activation-dependent intraplatelet signals. An outcome of GPVI engagement is the release of platelet-derived microparticles (arbitrarily defined as less than 1 μm) and, in the case of rheumatoid arthritis, the microparticle further augments the inflammatory cascade. Beyond this example, platelet-derived microparticles are being recognized as relevant in a wide range of proinflammatory and prothrombotic pathophysiologic states. Characterization of the human GPVI gene has revealed genetic haplotypes corresponding to altered in vitro platelet response. A growing body of evidence has also linked these haplotypes to pathologic responses in the acute setting, such as myocardial infarction and stroke. The GPVI haplotypes, referred to as GP6a and GP6b, differ in 5 codons resulting in 3 polymorphic amino acids in the extracellular portion of the GPVI (S219P, K237E, and T249A) and 2 polymorphic amino acids in the cytoplasmic tail (Q317L and H322N). While GPVI haplotypes have been linked to cardiovascular disease outcome, a gap exists in determining whether the same polymorphisms influence platelet microparticle production and whether microparticle levels might vary in disease states. We have performed a detailed flow cytometry analysis of GPVI-dependent platelet-derived microparticles both in vivo and following agonist stimulation in vitro using blood from normal donors. A correlation exists between GPVI haplotypes and circulating levels of platelet-specific (αIIb) microparticles. Comparing homozygous GP6a (n=34) and heterozygous GP6a/b (n=17) individuals reveals an approximate 2-fold difference in the number of platelet microparticles that is statistically significant (p= 0.005). The GPVI dependent in vitro release is also haplotype specific following addition of type I fibrillar collagen. However, in a human condition of chronic inflammation, such as rheumatoid arthritis, haplotype-dependent changes in platelet-derived microparticle are not observed and reveal a maximum level of microparticle release occurring in the chronic inflammatory state. The results demonstrate that common polymorphisms within the human GPVI gene impact the level of circulating platelet-specific microparticles in normal individuals and in a state of chronic inflammation. The physiologic contribution of GPVI in disease states must consider the representative haplotype and levels of GPVI-dependent microparticles.

Disclosures:
No relevant conflicts of interest to declare.
Effects Of Helicobacter Pylori Eradication In Adult Patients With Acute Idiopathic Thrombocytopenic Purpura (ITP)
Authors: Moghimi Minoosh , Dizaji Saeid Razavi , Mortazavi Yousef , Mazloomzadeh Saeideh , Keyhanian Shaharbano , Alamsamimi Mozhgan 
Published: October 2019

Abstract

Background
Different studies have investigated the link between helicobacter pylori and extra digestive diseases suchas ITP.

Aims
In most studies chronic ITP has been investigated, therefore in the present study, we focused on the effect of H.pylori eradication on chronicity of ITP in adult patients with acute ITP.

Methods
After obtaining informed consent, eighty five patients with acute ITP whose platelet count were less than 30 ×103 /μl were enrolled to the study. Urea breath test (UBT) was carried out for all the patients. Based on the UBT results,the patients were divided into 3 groups: 1- H.pylori positive patients who underwent standard triple therapy. 2- ITPpatients negative for H.pylori and 3- ITP patients positive for H.pylori, but without eradication therapy. Evaluation of the H.pylori eradication was done 4 weeks after the treatment and also at the end of month 6.

Results
52(61.2%) of patients were female and 33(38.8%) were male with mean age of 34.8±12.2 years. There was nosignificant difference between the mean age and sex in different groups. No significant difference was seen in mean platelet count at the baseline among the groups, but there was a significant statistical difference in mean platelet betweenthe groups at the end of the first month. However, a significant difference was not seen in mean platelet count in months 2to 6. Chronicity in the first group was less than the 3rd group which statistically was significant. Also, the chronicity rate in non-infected ITP patients was lower than the H.pylori positive patients (P=0.03).

Summary / Conclusion
Our results show that eradication of H.pylori can reduce the chronicity rate in adult patients with acute ITP. Further studies on larger number of patients with longer follow-up are recommended.

Disclosures:
No relevant conflicts of interest to declare.
The Dynamic Motion and Delicate Control Of Vkor Catalysis
Authors: Li Weikai 
Published: October 2019

Vitamin K epoxide reductase (VKOR) is an intramembrane enzyme required for blood coagulation and is the target of the anticoagulant warfarin. The reduction of vitamin K epoxide is coupled with disulfide-bond formation at the active site of VKOR. To regenerate the active site, VKOR is reduced by protein partners that transfer electrons to VKOR. Here we report two crystal structures of a bacterial VKOR homolog with its reducing partner captured in different conformational states. These structures reveal a short helix at the hydrophobic active site of VKOR that undergoes stretching and compressing motions. Motions of this “horizontal helix” promote electron transfer to the VKOR active site by regulating the positions of two cysteines in an adjacent loop. Compression of the helix also separates the “loop cysteines” to prevent backward electron transfer. During these motions, hydrophobicity at the active site is maintained to facilitate vitamin K reduction. Biochemical experiments support the structural model and suggest that several warfarin-resistant mutations act by changing the conformation of the horizontal helix. Taken together, these structures provide a comprehensive understanding of electron transfer and vitamin K reduction catalyzed by VKOR.

Disclosures:
No relevant conflicts of interest to declare.
Trib2 Pseudokinase Marks Early CMP Progenitors, and Promotes Granulocytic Plus Erythroid Progenitor Cell Formation
Authors: Li Lei , Kuhrt David , Dev Arvind , Jachimowicz Edward , Akashi Koichi , Wojchowski Don M. 
Published: October 2019

Abstract
Altered expression of Trib2 pseudokinase has been associated with AML (M2, M4 subtypes) (Cancer Cell, 10: 401), and ectopic expression of Trib2 in hematopoietic progenitors drives myeloid leukemogenesis (Blood, 116: 1321). In addition, inhibition of Trib2 expression by EPO in erythroid progenitors has been demonstrated (Blood, 111: 5390). Beyond this, little is known about Trib2’s expression profiles and roles during normal myelopoiesis. To critically address functional roles, we have generated and characterized a novel Trib2-KO mouse model. Within bone marrow CFU-GM formation faltered, and Trib2-KO Linneg progenitors exhibited deficit granulocyte production (with a relative increase in monocytes). This was not associated with significant differences in survival or cell cycle features among developing granulomonocytic cells. Instead, Trib2 levels proved to be heightened among Linneg progenitors (and inhibited in GM cells following GMCSF, GCSF or ATRA exposure). This prompted analyses of CMP and early-CMP (eCMP) populations in which RNAseq demonstrated peak Trib2 expression. Furthermore, eCMP and CMP pools were elevated in Trib2-KO mice. Within progenitors of the erythroid lineage, Trib2 expression levels also selectively increased, and in Trib2-KO mice BFUe and CFUe levels were decreased (and RBC plus HGB were diminished). Intriguingly, when Trib2-KO mice were challenged with Flt3L plus GMCSF, BFUe (but not CFU-GM or GEMM) were mobilized to peripheral blood at levels 13.3-fold above wild-type controls. This indicates candidate effects of Trib2 on a proposed BFUe niche. Overall, novel multi-lineage roles for Trib2 are revealed in regulating eCMP pools, GM-CSF populations, balanced granulocytic vs monocytic cell formation, and the production of erythroid progenitor cells.

Disclosures:
No relevant conflicts of interest to declare.
Loss Of NIPA Leads To Defects In The Repair Of DNA Double Strand Breaks In Mice
Authors: Illert Anna Lena , Antinozzi Cristina , Kawaguchi Hiroyuki , Kulinski Michal , Klitzing Christine , Peschel Christian , de Rooji Dirk , Morris Stephan w. , Barchi Marco , Duyster Justus 
Published: October 2019

Abstract
Regulated oscillation of protein expression is an essential mechanism of cell cycle control. The SCF class of E3 ubiquitin ligases is involved in this process by targeting cell cycle regulatory proteins for degradation by the proteasome. We previously reported the cloning of NIPA (Nuclear Interaction Partner of ALK) in complex with constitutively active oncogenic fusions of ALK, which contributes to the development of lymphomas and sarcomas. Subsequently we characterized NIPA as a F-Box protein that defines an oscillating ubiquitin E3 ligase targeting nuclear cyclin B1 in interphase thus contributing to the timing of mitotic entry.
Using a conditional knockout strategy we inactivated the gene encoding Nipa. Nipa-deficient animals are viable, but show a lower birth rate and a reduced body weight. Furthermore, Nipa-deficient males were sterile due to a block of spermatogenesis during meiotic prophase. Virtually no spermatocytes progress beyond a late-zygotene to early-pachytene stage and reach an aberrant stage, with synaptonemal complex disassembly and incomplete synapsis. Nipa-/- females are sub-fertile with an early and severe meiotic defect during embryogenesis with extensive apoptosis in early prophase (E13.5-E14.5).
Here we report, that Nipa-/- meiocytes exhibit persistent cytological markers for DNA double strand break repair proteins (like DMC1, RAD51) in meiotic prophase with more than twice as many DMC1 foci as control animals. Kinetic analysis of the first wave of spermatogenesis showed increased DMC1/RAD51 foci in Nipa-/- cells as soon as early-pachynema cells appear (13-14 days post partum). Moreover, we show that Nipa deficiency does not lead to a defect in meiotic sex chromosome inactivation despite epithelial stage IV apoptosis. Nipa-deficient spermatocytes exhibit numerous abnormalities in staining of chromosome axis associated proteins (like SYCP3 and STAG3) indicating that chromosome axis defects were associated with compromised chromosome axis integrity leading to overt chromosome fragmentation.
Further in vitro analyses with bleomycin treated MEFs displayed high pH2AX levels in cells lacking NIPA. Repair of DNA DSB seemed to be abolished in these cells as the pH2AX-level were sustained and still visible after 90 min of timecourse, where wildtype cells already repaired sides of DNA Damage. Consistent with these findings NIPA-deficient spleen cells showed compromised DNA Damage repair measured in a comet assay with a significantly longer olive tail moment in NIPA knockout cells under repair conditions.
Taken together, the phenotype of Nipa-knockout mice is a definitive proof of the meiotic significance of NIPA and our results show a new, unsuspected role of NIPA in chromosome stability and the repair of DNA double strand breaks.

Disclosures:
No relevant conflicts of interest to declare.