bluuebunny/crossref_metadata_2025_split huggingface dataset:
Treatment of Serum Ferritin Deficiency Regardless Haemoglobin Level
Authors: Al-Jafar Hassan A. 
Published: October 2019

Abstract
Abstract 5166

Introduction
The concentration of serum ferritin is positively correlated with the size of the total body iron stores in the absence of inflammation. [1] It is mainly concentrated in the liver, spleen and bone marrow in humans. The serum ferritin is present in all cells of the body. [ 2] Normal values are generally 12 – 300 ng/mL for men and 12 – 150 ng/mL for women. Some patients have clinical symptoms of iron deficiency anaemia as fatigue, general weakness, hair loss, depressed mode although their haemoglobin and serum ferritin are in the lower normal values [3].

Method
Patients sample are 1- females 2- aged from 15–70 years 3-who have problems with oral iron either not getting benefit from it or can't tolerated it or not compliant with oral iron treatment. On presentation patients subjected for the following tests and then repeated after one month of intravenous iron infusion 1- complete blood count, 2- serum ferritin, 3- serum vitamin B12 and folate, 4- HPLC, 5-G6PD, 6- Coagulation profile (INR, APTT), 7 – LFT and RFT.

Results
Group 1: responded laboratory and clinically to treatment.
Group 2: responded laboratory and clinically to treatment.
Group 3: responded laboratory and clinically to treatment.

Discussion
Three group of patients noticed:
Group 1 Low Hb and low s. ferritin where symptoms of iron deficiency are exist in this group of patients.
Group 2 Hb is normal and s. ferretin where symptoms of iron deficiency are exist in this group of patients.
Group 3 both Hb and s. ferretin where symptoms of iron deficiency are exist in this group of patients.
Group 4 control group was just observed and reassured.
Iron is essential element in all cells of human [4], so when it is low it means dysfunctions in other cells and organs like hair, muscles and brain.

Conclusion
Lower normal level of Serum Ferritin is enough for normal Hb level but it is may be not enough for normal functions in other organs where the iron is needed. So that why it might be need to treat S. ferritin deficiency and shouldn't wait to reach to the level of IDA. According to that it might be indicated to: 1- Treat patients when s. ferritin is low even if Hb is normal. 2 - Lower normal value has to be increased to at least to 50mg to ensure that not only Hb will become normal but also other organ function.
Putting in mind that the range of Hb could reach to 305ng/ml without reaching to the toxic level of iron overload.

Reference

Aim
Is to assess the benefit of raise up Serum Ferritin level to a new higher reference lower value as 50 ng/mL instead of the present lower values, because the present lower normal value 11 ng/mL, may be enough to get haemoglobin to the normal level and function, but it seems not enough for normal functions of other different organs as the hair, muscles, brain, which could be the reason of hair lose, general weakness, fatigue, depressed mode, . ect although haemoglobin is normal and s. ferritin is low or in lower normal value.

Disclosures:
No relevant conflicts of interest to declare.
Thalassemia – Increasingly Frequent Cause of Microcytic Anemias in Children in Poland
Authors: Albrecht Katarzyna , Adamowicz-Salach Anna , Burzynska Beata , Pawelec Katarzyna , Matysiak Michal 
Published: October 2019

Abstract
Abstract 5184
Thalassemias are a group of genetically determined hemolytic anemias related to disturbed synthesis of globin chains, and are classified as quantitative hemoglobinopathies.
We would like to present our experience in diagnosis of thalassemia in children as an increasingly frequent cause of microcytic anemias as well as to point out that it is the cause of microcytic anemia in Poland.
Between 2005–2012 we established the diagnosis of thalassemia minor in 94 children of Polish origin, referred to the Department of Pediatrics, Hematology and Oncology, Medical University of Warsaw. At admission all patients presented microcytosis, erythrocytosis, hypochromia and normal or elevated iron status. Most of them had long-lasting uneffective iron therapy in anamnesis. Their age ranged from 3 to 17 years. The group consisted of 40 girls and 54 boys.
Routinely employed diagnostic methods, including blood morphology, determination of iron and ferritin, electrophoretic separation of hemoglobins, determination of HgbA2 and HgbF, as applied in all cases, often turn out to be insufficient to diagnose thalassemia, substantiating the need to use modern molecular biological techniques.
Thalassemia alfa was diagnosed in 9 children in whom we detected deletion α 3, 7 using molecular biological techniques.
In 2 children coexistence of both thalassemia alfa and beta was established. Both children presented macrocytosis and normal range of haemoglobin and red blood cells count. The molecular biological techniques used in both cases identified deletion α 3, 7 characteristic for alfa thalassemia, and Hb Koln (Codon GTC>ATG) and Del 4, 2 (Taq1) characteristic for beta thalassemia.
In the remaining 83 patients we made the diagnosis of thalassemia beta. The diagnosis was possible after the evaluation of Hgb A2 and F levels. In 24 cases we managed to performed molecular biological techniques, identifying mutation IVS I-6 (T>C) as the most frequent.
Generally associated with the Mediterranean Basin, thalassemia is, however, more and more often diagnosed in Poland. This is due to the international human traffic. Growing availability of diagnostic tests, like molecular biological techniques, especially useful in equivocal cases, is also important.
The amplification of diagnostic possibilities by genetic tests in the future may be useful in detection of genetic mutations responsible for thalassemia and characteristic for Polish population.

Disclosures:
No relevant conflicts of interest to declare.
The Role of STAT5 in FLT3-Mediated Leukemogenesis
Authors: Müller Tony Andreas , Dechow Rebekka , Istvanffy Rouzanna , Hennighausen Lothar , Peschel Christian , Duyster Justus 
Published: October 2019

Abstract
Abstract 771

Introduction:
Activating mutations of FLT3 are among the most frequent alterations in AML. Two distinct types of mutations have been described: Duplication of the juxtamembraneous domain (ITD) and point mutations of the tyrosine kinase domain (TKD). Although both lead to constitutive FLT3 signaling, FLT3-ITD signals through the STAT5 pathway whereas FLT3-TKD and FLT3 WT activate STAT5 to a much lesser degree. In a murine bone marrow transplantation model, FLT3-ITD leads to a myeloproliferative neoplasm; FLT3-TKD induces a lymphoid disease with significantly longer latency. Interaction of SRC with FLT3-ITD was recently shown to be necessary for STAT5 activation.

Methods:
Fetal liver cells from constitutive STAT5 knockout mice were infected with FLT3 oncogenes and cultured on methylcellulose. Conditional STAT5 knockout mice were analyzed for phenotype and latency of hematologic disorders after transplantation with FLT3-ITD or FLT3-TKD-infected bone marrow. Expression of STAT5 target genes was determined using Realtime-PCR. Phospho-STAT5 levels were quantified by intracellular staining and flow cytometry.

Results:
Depletion of STAT5 leads to a dramatic decrease in the ability of fetal liver cells to form colonies in Methylcellulose when infected with FLT3-ITD, but not when infected with FLT3-TKD. In-vivo studies using a murine conditional knockout model proved that STAT5 is necessary for the induction of a myeloproliferative disease by FLT3-ITD, while FLT3-TKD is not dependent on STAT5. Mice with deletion of the STAT5 allele displayed expansion of mainly lymphoid cells with an increased latency similar to the FLT3-TKD disease. In additional studies differential activation of STAT5 in myeloid and lymphoid progenitors was found. The STAT5 target gene Oncostatin M is exclusively expressed in myeloid progenitor cells infected with FLT3-ITD and in mice with intact STAT5 that were transplanted with FLT3-ITD. Furthermore, elevated SRC expression levels could be detected in proliferating myeloid progenitor cells compared to lymphoid progenitors.

Conclusion:
STAT5 is critical for the induction of a myeloproliferative disease by FLT3-ITD in a murine transplantation model. Interaction of SRC and FLT3-ITD leads to the activation of STAT5 in myeloid progenitors, but not in lymphoid progenitors expressing low levels of SRC. Oncostatin M upregulation is likely to be a downstream process contributing to the phenotype and latency of the disease.

Disclosures:
No relevant conflicts of interest to declare.
Impaired Neutrophil Function in 24p3 Null Mice Contributes to Enhanced Susceptibility to Bacterial Infections
Authors: Devireddy L.R. , Liu Zhuoming 
Published: October 2019

Abstract
Abstract 833
Lipocalin 24p3 (24p3) is a neutrophil secondary granule protein. 24p3 is also a siderocalin, which binds several bacterial siderophores. It was therefore proposed that synthesis and secretion of 24p3 by stimulated macrophages or release of 24p3 upon neutrophil degranulation sequesters iron-laden siderophores to attenuate bacterial growth. Accordingly, 24p3-deficient mice are susceptible to bacterial pathogens whose siderophores are chelated by 24p3. Specific granule deficiency (SGD) is a rare congenital disorder characterized by complete absence of proteins of secondary granules. Neutrophils from SGD patients lack normal functions and are prone to bacterial infections, but the importance of 24p3 for neutrophil dysfunction in SGD is not known. Here we show that neutrophils of 24p3−/− mice are defective for many neutrophil functions. Specifically, neutrophils in 24p3−/− mice do not extravasate to sites of infection and are defective for chemotaxis. A transcriptome analysis revealed that genes that control cytoskeletal reorganization are selectively suppressed in 24p3−/− neutrophils. Additionally, small regulatory RNAs (miRNAs) that control upstream regulators of cytoskeletal proteins are also increased in 24p3−/− neutrophils. Further, 24p3−/− neutrophils failed to phagocytose bacteria, which may account for the enhanced sensitivity of 24p3−/− mice to both intracellular (Listeria monocytogenes) and extracellular (Candida albicans, Staphylococcus aureus) pathogens. Interestingly, Listeria does not secrete siderophores and additionally, the siderophore secreted by Candida is not sequestered by 24p3. Therefore, the heightened sensitivity of 24p3−/− mice to these pathogens is not due to sequestration of siderophores to limit the iron availability but as a consequence of impaired neutrophil functions.

Disclosures:
No relevant conflicts of interest to declare.
p21-Activated Kinases Regulate Directional Migration and Cytoskeletal Organization in Human Neutrophils
Authors: Itakura Asako , Aslan Joseph E. , Kusanto Branden T. , Phillips Kevin G. , Insall Robert H. , Chernoff Jonathan , McCarty Owen J.T. 
Published: October 2019

Abstract
Abstract 834
Neutrophil chemotaxis is controlled by coordinated processes of directional sensing, polarization and motility. This study was designed to characterize the role of p21-activated kinases (PAKs) during the chemotaxis of human primary neutrophils. PAKs are known as effectors of the Rho GTPases Rac and Cdc42. It has been shown that PAK1 and PAK2 are strongly activated downstream of the f-Met-Leu-Phe (fMLP) receptor via Rac (Huang et al., MCB 1998). PAK1 is known to localize in lamellipodia at the leading edge of human neutrophils (Dharmawardhane et al., JLB 1999) and mediate persistent directional migration via Cdc42 in a neutrophil-like cell line (Li et al., Cell 2003). However, little is known about the specific role of PAK isoforms in spatial/temporal regulation of cytoskeletal dynamics in human neutrophils.
Our data show that human neutrophils express PAK1, 2 and 4. Under an fMLP gradient, human neutrophils developed morphological polarity with a distinct leading edge and rear, and migrated up the fMLP gradient at the speed of 7.5 ± 0.56 μm/min. Inhibition of Rac or PI3K impaired directionality but did not significantly affect migration speed of chemotaxing neutrophils (6.3 ± 0.56 μm/min or 6.2 ± 0.85 μm/min, respectively). In contrast, neutrophils treated with the PAK inhibitor, PF3758309 (PF), displayed random migration, less polarization and reduced motility (3.1 ± 0.21 μm/min). These results suggest that PAK regulates neutrophil chemotaxis independently of the Rac-PI3K axis. The presence of PF did not abrogate intracellular Ca2+mobilization in fMLP-driven chemotactic condition. Instead, the decreased migratory ability by PAK inhibition was associated with multiple Ca2+ spikes. Immunofluorescence imaging shows that PAK2 but not PAK1, was phosphorylated and translocated from cytosol to actin-rich leading edge in the proximity to GTP-bound Rac within 3 min of fMLP stimulation. Notably, PF treatment resulted in partial neutrophil spreading and actin/myosin II translocation in the absence of extracellular stimuli, suggesting that basal level of PAK phosphorylation may be required for cytoskeletal integrity of resting neutrophils. Neutrophils pretreated with PF displayed less activation and translocation of PAK2 and Rac.
In summary, our data demonstrate for the first time the distinct roles of PAK isoforms in human neutrophil morphological polarity and directional migration and suggest that PAK2 is activated downstream of fMLP receptor through Rho-family small GTPases.

Disclosures:
No relevant conflicts of interest to declare.
The Red Cell: How Erythropoiesis Modulates Hepcidin Expression
Authors: Ginzburg Yelena 
Published: October 2019

Abstract
Abstract SCI-24
Erythroid precursors in the bone marrow require transferrin-bound iron for hemoglobin synthesis. Therefore, it is not surprising that the regulation of erythropoiesis and iron metabolism is interlinked. Iron demand for erythropoiesis is communicated to the iron-regulatory machinery through incompletely understood mechanisms. At the core of systemic iron homeostasis is the peptide hormone hepcidin, restricting cellular iron export to plasma by inducing the endocytosis and proteolysis of ferroportin. Hepcidin, a liver-synthesized peptide hormone, is released in response to increased iron load, and there is early evidence that circulating hepcidin concentrations affect the distribution of iron between the macrophage storage compartment (favored by higher hepcidin concentrations) and parenchymal cells, including cardiac myocytes and hepatocytes (favored by low hepcidin). Furthermore, ferroportin has recently been identified on erythroid precursors. Its purpose in this cell type and its function in the interface between erythropoiesis and iron metabolism are unclear. Additionally, in response to bleeding or the administration of erythropoietin, expansion of erythroid precursors suppresses hepcidin, most likely through one or more mediators released by the bone marrow and acting on hepatocytes. Iron-loading anemias with ineffective erythropoiesis, in particular β-thalassemia, demonstrate the effects of pathological “erythroid regulators” of hepcidin. Although erythrocyte transfusions are the main cause of iron loading in patients who receive them (β-thalassemia major), lethal iron overload is seen also in patients who are rarely or never transfused (β-thalassemia intermedia). Here, iron hyperabsorption is the cause of iron overload and, as in hereditary hemochromatosis, is caused by low hepcidin. Decreased hepcidin expression in β-thalassemia, with concurrent ineffective erythropoiesis and iron overload, indicates that the “erythroid regulator” may play an even more substantial role in iron metabolism than the “stores regulator.” Two members of the bone morphogenetic protein (BMP) family, growth differentiation factor (GDF) 15 and Twisted Gastrulation (TWSG1), have been implicated as candidate bone marrow-derived hepcidin suppressors in β-thalassemia. Neither factor is responsible for the physiologic hepcidin suppression in response to hemorrhage-induced stress erythropoiesis, and the physiologic suppressor is not known. We focus here on the current state of knowledge regarding the regulation of iron metabolism and attempt to elucidate the interface between iron regulation and erythropoiesis using evidence in part derived from animal models of β-thalassemia. A more complete understanding of the coregulation of erythropoiesis and iron metabolism may lay the foundation for improving diagnosis, increasing treatment options, and ultimately impacting the well-being of patients afflicted with different anemias and/or iron overload related-disorders.

Disclosures:
No relevant conflicts of interest to declare.
Mechanisms of Transcriptional Regulation and Transformation by HOXA9
Authors: Hess Jay L. , Collins Cailin , Bronstein Joel , Sun Yuqing , Nagaraja Surya 
Published: October 2019

Abstract
Abstract SCI-30
HOXA9 plays important roles in both development and hematopoiesis and is overexpressed in more than 50 percent of acute myeloid leukemias (AML). Nearly all cases of AML with mixed lineage leukemia (MLL) translocations show increased HOXA9 expression, as well as cases with mutation of the nucleophosmin gene NPM1, overexpression of CDX2, and fusions of NUP98. In most cases, upregulation of HOXA9 is accompanied by upregulation of its homeodomain-containing cofactor MEIS1, which directly interacts with HOXA9. While HOXA9 alone is sufficient for transformation of hematopoietic stem cells in culture, the addition of MEIS1 increases the transformation efficiency and results in rapidly fatal leukemias in transplanted animals. Despite the crucial role that HOXA9 plays in development, hematopoiesis, and leukemia, its transcriptional targets and mechanisms of action are poorly understood. We have used ChIP-seq to identify Hoxa9 and Meis1 binding sites on a genome-wide level in myeloblastic cells, profiled their associated epigenetic modifications, identified the target genes regulated by HOXA9 and identified HOXA9 interacting proteins. HOXA9 and MEIS1 cobind at hundreds of promoter distal, highly evolutionarily conserved sites showing high levels of histone H3K4 monomethylation and CBP/P300 binding. These include many proleukemogenic gene loci, such as Erg, Flt3, Myb, Lmo2, and Sox4. In addition, HOXA9 binding sites overlap a subset of enhancers previously implicated in myeloid differentiation and inflammation. HOXA9 binding at enhancers stabilizes association of MEIS1 and lineage-restricted transcription factors, including C/EBPα, PU.1, and STAT5A/B thereby promoting CBP/p300 recruitment, histone acetylation, and transcriptional activation. Current efforts are focused on using both biochemical and genetic approaches to assess the role of HOXA9 “enhanceosome” components C/EBPα, PU.1, and STAT5A/B in transcriptional regulation and leukemogenesis. Studies to date suggest that C/EBPα and PU.1 binding can occur in the absence of HOXA9/MEIS1, supporting a model in which these proteins act as pioneer transcription factors for establishment of poised, but not activated, HOXA9-regulated enhancers. Work is under way to assess the impact of high-level HOXA9 and MEIS1 on enhanceosome assembly and the role of recruitment of transcriptional coactivators involved in target gene up- or downregulation, including histone acetyltransferases and chromatin remodeling complexes. Collectively, our findings suggest that HOXA9-regulated enhancers are a fundamental mechanism of HOX-mediated transcription in normal development that is deregulated in leukemia.

Disclosures:
No relevant conflicts of interest to declare.
In Vitro Production of Erythrocytes
Authors: Douay Luc 
Published: October 2019

Abstract
Abstract SCI-39
The generation of red blood cells (RBCs) in vitro using biotechnologies could represent an interesting alternative to classical transfusion products, in that it would combine adequate supplies with the specific production of blood products of a particular phenotype and the reduction of infection risks. This presentation will review how it is now possible to obtain in vitro complete maturation of the erythroid line to the stage of enucleation, starting from hematopoietic stem cells (HSCs) from peripheral blood, bone marrow or umbilical cord blood, or embryonic stem cells or adult pluripotent stem cells (induced pluripotent stem cells, iPSCs). This presentation will discuss how the functionality of cultured human RBCs (cRBCs) is settled in terms of deformability, hemoglobin maturation, oxygen carrying capacity, enzyme content, and terminal maturation from the reticulocyte stage to mature RBC after infusion into the NOD/SCID mouse model. The clinical feasibility of this concept has recently been demonstrated by reporting that cRBCs generated in vitro from peripheral HSCs under GMP conditions encounter in vivo the conditions required for their maturation and that they persist in the circulation for several weeks in humans. These data have established the proof of principle for transfusion of in vitro-generated RBCs and the pathway toward new developments in transfusion medicine. The most proliferative source of stem cells for generating cRBCs is cord blood, but it is limited in terms of HSCs and is dependent on donations. Pluripotent stem cell technology represents a potentially unlimited source of RBCs and opens the door to the development of a new generation of allogeneic transfusion products. Because iPSCs can be selected for a phenotype of interest, they are obviously the best candidate for organizing complementary sources of RBCs for transfusion. It is established that only three human iPSC clones would have been sufficient to match more than 99 percent of the patients in need of RBC transfusions. As a whole, a very limited number of RBC clones would provide for the needs of most alloimmunized patients and those with a rare blood group. Generating cRBCs from iPSCs has been done but needs to be optimized to lead to a clinical application in blood transfusion. Several crucial points remain to be resolved, notably, the choice of the initial cell type, the method of reprogramming (i.e., to ensure the safety of the iPSCs and to ensure their clinical grade), the optimization of the erythrocyte differentiation, and the definition of GMP conditions for industrial production. Assuming that in vitro large-scale cultured RBC production efficiently operates in the near future, this presentation will highlight the potential applications for alloimmunized patients and those with a rare blood group.

Disclosures:
No relevant conflicts of interest to declare.