bluuebunny/crossref_metadata_2025_split huggingface dataset:
Angelica Polysaccharides Promotes Thrombopoiesis through the Phosphatidylinositol 3-Kinase/Akt Pathway.
Authors: Yang Mo , Chan Godfrey Chifung , Ye Jie Yu , Liu Chang 
Published: April 2020

Abstract
Angelica Polysaccharide is from the root of Radix Angelicae Sinensis (Chinese Danggui). Danggui has been used for centuries to treat blood-deficiency related diseases. The hematopoietic effect of Danggui may be related to its constituent, polysaccharide. However, the effects of angelica polysaccharide on hematopoietic stem/progenitor cells and megakaryocyte/platelet lineage have not been well studied. In this study, we specifically investigate the thrombopoietic effect of Angelica polysaccharide (APS) in a mouse model and its molecular mechanism. A myelosuppression mouse model (4-Gy-irradiated) was treated with APS (10 mg/kg/day) and thrombopoietin (TPO) (1 mg/kg/day). Peripheral blood cells from APS, TPO and vehicle-treated samples were counted on days 0, 7, 14 and 21. Then CFU assays were used to determine the effects of APS on the megakaryocytic progenitor cells and other lineages. Analyses of Annexin V, Caspase-3, and Mitochondrial Membrane Potential were conducted in megakaryocytic cell line M-07e. The effects of APS on cells treated with Ly294002, a Phosphatidylinositol 3-Kinse inhibitor and the effect of APS on the phosphorylation of AKT were also studied. APS as well as TPO significantly enhanced the recovery of platelet and WBC count, and bone marrow CFU-MK and CFU-GM formation (n=6). Morphological examination of bone marrows showed that APS treatment significantly increased the recovery of the megakaryocytic series. APS also increased the recovery of the body weight and organ weight (liver and spleen). However, APS didn’t have a up-regulating effect on TPO production by a ELISA assay. We further analyzed the in vitro effect of APS on CFU-MK and CFU-GM formation. APS (50 ug/ml) enhanced TPO (50 ng/ml) -induced CFU-MK (p=0.06, n=4), and CFU-GM formation (p=0.032, n=6). However, APS alone (00 ug/ml) did not show significant effect on CFU-MK and CFU-GM proliferation (n=6). The effect of APS (1–500 ug/ml) on the growth of bone marrow stromal cells was further investigated using CFU-F (fibroblastoid) assay. APS (50 ug/ml) alone showed a promoting effect on CFU-F growth (p=0.049, n=3). APS (50 ug/ml) also significantly enhanced PDGF, bFGF and VEGF-induced CFU-F formation (n=4). Moreover, the anti-apoptotic effect of APS in M-07e cells was also demonstrated by using Annexin-V, Caspase-3, and JC-1 assays. Addition of Ly294002 alone increased the percentage of cells undergoing apoptosis. However, additional of APS to Ly294002-treated cells reversed the percentage of cells undergoing apoptosis. Furthermore, addition of APS significantly increased the phosphorylation of AKT. APS promotes thrombopoiesis in a mouse model. This effect is likely to be mediated by the PI3K/AKT pathway.
Blocking of IL-21 Signal Attenuates Graft-Versus-Host Disease but Not Graft-Versus-Leukemia Effect in a Mouse Model
Authors: Meguro Akiko , Ozaki Katsutoshi , Oh Iekuni , Matsu Haruko , Hatanaka Keiko , Tatara Raine , Leonard Warren J , Sato Kazuya , Ozawa Keiya 
Published: April 2020

Abstract
Immunological effects of interleukin-21 (IL-21) on T-, B-, and NK-cells have been reported, but the role of IL-21 in graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) remains obscure. Here, we demonstrate that splenocytes from IL-21R−/− mice show attenuated GVHD symptoms as compared to splenocytes from wild type mice. In addition, the survival of mice receiving IL-21R−/− splenocytes is significantly longer than of those receiving wild type splenocytes. Both types of splenocytes appear to efficiently eliminate leukemic cells, suggesting the retention of GVL effects. Our results suggest that CD4 T-cells augment GVHD by autocrine stimulation via IL-21, and that production of IL-17 is downregulated in IL-21R−/− splenocyte recipients. To develop a treatment model in mice, we generated a decoy receptor for IL-21, in which most of the intracellular domain is deleted. Mice receiving bone marrow cells transduced with this decoy receptor survived longer than those receiving cells transduced with an empty retrovirus.These results suggest that IL-21 promotes GVHD and that the blockade of IL- 21 signaling might be a useful treatment for GVHD, without causing loss of the GVL effect.
Immune Response of Human Polymorphnuclear Neutrophils to Aspergillus Fumigatus
Authors: Loeffler Juergen , Wozniok Iwona , Mezger Markus , Einsele Hermann 
Published: April 2020

Abstract
Invasive fungal infections with Aspergillus fumigatus show an increasing incidence due to the growing number of severely immunocompromised patients. Polymorphonuclear neutrophils (PMNs), as part of the innate immune system, are key players in antifungal immune responses, recognizing fungal pathogens at an early step of infection. Besides phagocytic mechanisms, PMNs kill pathogens by the release of reactive oxygen species (ROS). Human PMNs were isolated from blood of healthy donors using Biocoll separation. Cells were co-cultivated with conidia, germlings and hyphae of the clinical strain ATCC 9197. The oxidative burst was determined in a kinetic measurement quantifying dichlorfluorescein production. Chemotaxis was analyzed by transwell assays. Furthermore, total RNA was extracted and gene expression profiling was performed using Affymetrix U133Plus2.0 arrays. We could demonstrate that A. fumigatus represents a strong stimulus releasing ROS, depending on the morphotype; germlings revealed high ROS release, whereas resting conidia showed low stimulation capacity. PMNs actively tracked germlings and directly attached to fungi as demonstrated by real-time microscopy. Transwell assays revealed that chemotaxis of PMN is strongly dependent on the fungal morphotype; PMN showed only weak chemotaxis in the presence of conidia, whereas in the presence of germlings, high chemotactic activity was achieved. Furthermore, after 6h co-cultivation of PMNs and A. fumigatus germ tubes, 195 (1.273) genes showed an at least 4fold (2fold) altered gene expression. We observed upregulation of genes (hemoxygenase, heat shock 70kDa protein, HSPA8, HSPA1B, HSP90AB1, Ferritin) involved in self-protection against radicals. Additionally, genes involved in inflammatory responses (IL-8, CCL3, CXCL2, IL1RN) were significantly upregulated. Luminex analysis was performed for TNF-a, IL-12, GM-CSF, IFN-γ, IL-6, IL-8, IL-10 and IL-1b to identify secreted cytokines, thereby confirming array data. In conclusion, A. fumigatus had substantial effects on the activation of human PMNs. Various defence strategies were initiated, including phagocytosis, ROS release and mobilization of other immune effector cells by secretion of chemoattractants.
Familial Hemophagocytic Lymphohistiocytosis Type IV: Possible Founder Effect in Syntaxin 11 Gene Common Mutation Endemic to Turkish Population
Authors: Balta Gunay , Caliskan Umran , Guler Elif , Patiroglu Turkan , Oner Ahmet F. , Gurgey Aytemiz 
Published: April 2020

Abstract
Familial Hemophagocytic Lymphohistiocytosis (HLH) is a rare, autosomal recessive, if untreated, fatal disorder of early childhood characterized mainly by persistent fever, hepatocyplenomegaly, cytopenia, hypertriglyceridemia, hyperferritinemia, defective natural killer cell activity and hemophagocytosis in bone marrow, lymph nodes, liver and spleen. Familial HLH is a genetically heterogeneous disease; at least four sub-types (I–IV) associated with three genes and one chromosomal locus (9q21.3-q22) have been reported in the disease to date. A number of mutations in both Perforin and Munc13– 4 genes have been described in different populations whereas only 3 different types of mutations in Syntaxin 11 gene have, thus far, been described in a few families solely from Turkish population. Therefore, elucidation of the mutation spectrum of the Syntaxin 11 gene mostly depends on the identification of its genetic defects in this population. In this communication, we present our findings on Syntaxin 11 gene which is responsible for the familial HLH type IV. A total of 84 potentially primary HLH patients selected on the basis of consanguinity and/or family history and/or relapse were included in the study. Patients and family members were studied by linkage analysis for all 4 described gene loci and homozygosity for Syntaxin 11 gene was observed in 20 patients from consanguineous families. Sequencing of these patients for the complete coding and flanking sequences of Syntaxin 11 gene revealed molecular defects in only 5 patients (2M/3F) with the ages ranging from 2 months to 3 years. Contrary to the heterogeneity in the mutation spectrum of hereditary disorders generally observed in Turkish population, a single type of homozygous mutation (c369–370delAG, c374–376delCGC) was identified in these 5 unrelated patients who had a family history of at least one deceased sibling except one. Screening of the family members revealed that the obligate carriers and some of the siblings were heterozygous for the mutation. This 2 plus 3 nucleotide deletion in exon 2 leads to frameshift and premature termination of translation 59 altered aminoacids downstream (Val124fsX60). Haplotype analysis indicated that these patients who were coming from different parts of Turkey may have a common ancestor (founder effect). Besides confirmation of the “Turkish population endemic” profile of the Syntaxin 11 gene mutations, the findings provided here elucidate the single/common mutation characteristic of the gene which is extremely important in the molecular diagnosis of the disease. This study was supported by TUBITAK (Project No: 105S386; SBAG-3193).
Functional Characterization of Signaling Events Underlying Immunostimulatory and Immunosuppressive Activity of Macrophages.
Authors: Kloss Mercedes , Kanz Lothar , Salih Helmut R , Krusch Matthias 
Published: April 2020

Abstract
Macrophages (Mo) play an important role in combating infectious pathogens and regulating adaptive immune responses. Their reactivity is, among others, guided by pattern recognition receptors including Toll like receptor 4 (TLR4), which recognizes bacterial lipopolysaccharide (LPS). After stimulation, TLR4 can associate with different adaptor proteins like e.g. MyD88 or TRIF leading to the activation of signaling molecules like MAPK or NF-κB. However, stimulation with LPS may result in equally impressive immunostimulatory and immunosuppressive Mo responses. The signaling events responsible for this diametrical behavior of different Mo subsets are yet not fully understood. To study the effects of LPS on Mo we differentiated monocytes of healthy donors with GM-CSF and M-CSF into pro-inflammatory type 1 (Mo1) or anti-inflammatory type 2 (Mo2) subsets, respectively. As expected, upon LPS stimulation the pro-inflammatory Mo1 secreted low levels of the immunosuppressive cytokine IL-10 and high levels of IL-12, while the anti-inflammatory Mo2 produced neglectable amounts of IL-12 but high levels of IL-10. To elucidate the underlying molecular mechanisms we performed gene array analyses with Mo1 compared to Mo2 from three independent donors. Interestingly, among over 50 genes involved in TLR4 signaling, only TLR4 itself and its adaptor protein MyD88 were found to be differentially regulated: Within the immunosuppressive Mo2 subset, MyD88 was significantly downregulated, while a significant upregulation of TLR4 was observed compared to the immunostimulatory Mo1 subset. The differential expression pattern were further confirmed on protein level by western blot. Since TLR4 stimulation leads to MyD88-dependent activation of the MAPK JNK, we investigated whether JNK was differentially involved in cytokine production of Mo1 and Mo2. Addition of a specific JNK inhibitor (SB600125) concentration dependently reduced TLR4-induced IL-10 but enhanced IL-12 production in Mo1, pointing to an immunoinhibitory role of JNK in Mo1. In contrast, inhibition of JNK did not alter the secreted levels of IL-12 or IL-10 in Mo2, indicating that rather MyD88-independent pathways are responsible for the anti-inflammatory behavior of Mo2. In conclusion, our data suggest that the opposite regulation of the adaptor molecule MyD88 in pro- and anti-inflammatory Mo subsets is responsible for the fact that a single, non-polymorphic receptor like TLR4 can mediate both pro- and anti-inflammatory immune responses in Mo.
Patient-Reported Fatigue Scores in Cancer Patients with Chemotherapy-Induced Anemia Treated with Erythropoietic-Stimulating Agents (ESAs).
Authors: Burton Tanya , Hoaglin David , Larholt Kay , Apgar Elizabeth , Pashos Chris , Bookhart Brahim , Corral Mitra , Piech Catherine Tak , McKenzie R. scott 
Published: April 2020

Abstract
Background: Clinical trial evidence has demonstrated greater anemia-related fatigue at lower hemoglobin (Hb) levels in ESA-treated patients with chemotherapy-induced anemia (CIA). To understand these patient-reported outcomes (PROs), during ESA treatment in a real-world setting, data were analyzed from the Dosing and Outcomes Study of Erythropoiesis-Stimulating Therapies (D.O.S.E.) Registry, an ongoing, prospective, observational study.
Methods: Patients included for analysis were required to have a baseline Hb prior to ESA administration and at least two ESA administrations. PROs were measured using the Functional Assessment of Cancer Therapy: Fatigue subscale (FACT-F), a 13-question measure of anemia-associated fatigue with lower scores indicating greater fatigue in patients with CIA. Patients were to have completed up to three FACT-F questionnaires within seven days of a post-baseline Hb level. If multiple Hb values were identified, the Hb value closest to the FACT-F assessment was reported. The association between FACT-F scores and Hb levels was tested in a repeated-measures analysis. Mean FACT-F scores were calculated for specified Hb ranges.
Results: 740 CIA patients from 63 sites were identified between 12/03 and 7/08. 1088 concurrent FACT-F and Hb level assessments were available during ESA treatment. Mean FACT-F scores were lower when patients had lower Hb levels (table). The association between FACT-F scores and Hb levels showed a significant positive trend (p&lt;.0001).
Hb ≤ 8.0 g/dL 8.01–9.0 g/dL 9.01–10.0 g/dL 10.01–11.0 g/dL 11.01–12.0 g/dL n (concurrent FACT-F and Hb assessments) 19 63 212 336 458 Mean FACT-F score (SD) 21.26 (11.01) 22.95 (12.44) 25.64 (12.40) 26.84 (11.70) 29.34 (12.04)
Conclusions: This analysis of FACT-F assessments in CIA patients suggests that lower Hb levels were associated with greater anemia-related fatigue. These findings are consistent with clinical trial evidence and provide observational PRO data to patients and providers regarding anemia-related fatigue during ESA treatment.
Unrelated Cord Blood Transplantation (UCBT) for Transfusion- Dependent Thalassemia a CIBMTR Audited Retrospective Analysis of 30 Consecutive Patients from a Single Center
Authors: Jaing Tang-Her , Wang Brian , Gjertson David , Law Ping , Petz Lawrence , Chow Robert 
Published: April 2020

Abstract
UCBT offers a cure for thalassemia and has advantages as a stem cell source because the requirement for an HLA match between donor and recipient is less stringent. Previous reports of UCBT for thalassemia have yielded transplant related mortality (TRM) that was quite high and it is known that pre-freeze total nucleated cell (TNC) dose is critical for UCBT. With strategies that maximize TNC dose – using non-red cell reduced but plasma depleted (PD) CB, no post-thaw wash (NW), and double cord transplantation (DCT) when necessary – we have achieved promising results with UCBT in young patients with transfusion-dependent β-thalassemia. Between 10/2003 and 5/2008, 39 CB products were infused after Bu/Cy/ATG myeloablation into 30 pediatric thalassemia major patients (9 DCT) using NW PD CB exclusively. Patient status: 21 Pesaro class−1, 8 Pesaro class−2, and 2 unknown. Median age was 5 years (range 1–14 years) with a median weight of 18 kg (range 11–37 kg). The data was audited by the transplant center (TC) and on-site by CIBMTR (98.5% accuracy for Chang Gung). Median pre-freeze TNC cell dose was 10.9×107/kg, and median pre-freeze CD34+ cell dose was 4.0×105/kg. The number of CB products HLA ABDR matched were 4, 11, and 24 for 6/6, 5/6, and 4/6 or fewer matches. No significant adverse events were observed despite major ABO incompatibility in some cases and forgoing post-thaw wash. Cumulative incidences of donor-derived (determined by chimerism studies) neutrophil (ANC500), platelet 20K and 50K (plt 20K & 50K) engraftment were 96±11%, 92±11% and 92±12%, and median times to ANC500, plt 20K and 50K engraftment were 17.5 days (range 11–26), 49.5 days (range 28–135) and 63.5 days (range 35–203), respectively. Grade III/IV acute GvHD occurred in 40±9%, and extensive chronic GvHD occurred in 4±4% of the patients. Transplant related mortality were 10±5% at 100 days and 13±6% at 1 year. Overall survival at 1 and 3 years were 87±6% and 82±8%, while disease-free survival at 1 and 3 years were 85±7% and 78±9%. Five patients expired including 1 death prior to day +10 due to traumatic head injury. Mean and median follow up times were 24 and 16 months respectively (range 0.3–58 months) as of August 2008. To our knowledge, this is the largest single-institutional UCBT series for thalassemia, and shows the favorable clinical results that are attainable when TNC dose is optimized with PD CBU, no post-thaw wash and DCT when necessary.
Participation in a Clinical Trial Does Not Impact Outcome in Pediatric Acute Lymphoblastic Leukemia
Authors: Koschmann Carl , Hawkins Doug S , Thomson Blythe 
Published: April 2020

Abstract
Background: Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy. Over the past four decades, the outcome for pediatric ALL has rapidly improved secondary to the participation of children on clinical trials, resulting in standardization of the treatment. Some have argued that the best outcome results from participation in a pediatric clinical trial. To address the potential benefits and barriers to participation in a clinical trial, we analyzed clinical trial participation in pediatric ALL at a large single institution.
Methods: We evaluated 322 ALL patients &lt; 22 years old at diagnosis who receive their initial therapy at Seattle Children’s Hospital Regional Medical Center (S-CHRMC) between January 1997 and December 2005. Using a retrospective chart review, we analyzed the following variables: study participation (SP) or non-participation (NP) in an ALL therapeutic study, gender, race, patient immunophenotype, risk group (standard risk (SR), high risk (HR) or infant), home state, and distance of primary residence from S-CHRMC. S-CHRMC is the largest pediatric cancer center in the Pacific Northwest, with referrals from Washington, Alaska, Montana, Idaho and Wyoming. Events were defined as relapse or death from any cause.
Results: The overall 5 year event free survival (EFS) was 78% (+/− 2.5%). 157 patients participated in a treatment research study (49%). Only risk group was associated with EFS. SP and NP had similar EFS. Gender, race, immunophenotype, home state, or distance from primary residence were not associated with outcome. There was no difference in study participation by gender, race, home state, or distance of primary residence. There were trends to increased participation in SR vs HR (54% vs 35%, p value 0.15) and B lineage vs T lineage (50% vs 35%, p value 0.11).
Variable n 5 year EFS p value Study participation No 165 76 (+/− 3.9) 0.47 Yes 157 81 (+/− 3.3) Risk group SR 175 85 (+/− 3.0) HR 132 73 (+/− 4.2) (0.038 to SR) Infant 15 59 (+/−12.9) (0.011 to SR)
Discussion: Participation in a research study for treatment of pediatric ALL was not associated with improved outcome in our large single institution series. Study participation was not different by clinical features, including distance to the primary residence. Strict standardization of treatment for all patients may contribute to the similar outcome for SP and NP.
Altered Erythroid and Megakaryocytic Differentiation in Mice Expressing a Unique Chromatin Remodeling Domain of Erythroid Krüppel-Like Factor (EKLF)
Authors: Jansen Valerie M. , Abramova Tatiana , Shim Eun-Hee , Ramachandran Shaji , Desgardin Aurelie , Parekh Vishwas , Jane Stephen , Cunningham John M 
Published: April 2020

Abstract
The zinc finger-encoding transacting factor EKLF binds key regulatory elements of many erythroid-specific genes, and is essential for definitive erythropoiesis. Mice lacking this factor (EKLF−/−) die of anemia by E15.5 of gestation, failing to activate β-globin gene transcription, and demonstrating a block in the erythroid differentiation program at the primitive erythroblast stage. In contrast, megakaryocytic progenitors are amplified in EKLFnull embryos, with increased Fli-1 gene expression (a marker of early megakaryocytic differentiation), consistent with the idea that EKLF modulates the megakaryocyticerythroid (M-E) differentiation switch. We have demonstrated that an amino terminal mutant of EKLF (Δ221EKLF), is required to induce chromatin remodeling at the β-globin promoter in an EKLF-null erythroid cell line, but additional amino terminal sequences are required for initiation of β-globin gene transcription (Brown et al., 2002). To evaluate the role of this chromatin remodeling (CR) domain in erythroid and megakaryocytic differentiation in vivo, we have generated a knock-in allele of EKLFCR allele. Similar to EKLF-null embryos, mice homozygous for this mutant allele die of anemia by E15.5 of gestation. In contrast to erythroid cells lacking EKLF, EKLFCR/CR progenitors demonstrate
appropriate binding of the CR encoding domain to all EKLF-regulatory sequences; a block in erythropoiesis at a more a mature stage in differentiation a chromatin architecture and histone modification pattern at erythroid-specific genes that recapitulates the events observed in EKLF+/+ erythroblasts at a similar stage of erythroid ontogeny; a failure of terminal erythroid gene transcription.
Examining the role of EKLFCR in megakaryopoiesis, we observed
inhibition of megakaryocytic progenitor amplification in EKLFCR/CR fetal hematopoietic cell populations when compared to EKLF-null embryos; loss of Fli-1 gene expression in EKLFCR expressing cells; binding of the EKLFCR mutant protein to the Fli-1 promoter with inhibition of gene transcription; a repressed chromatin architecture at megakaryocytic gene loci.
In contrast to these results, mice homozygous for a knockin allele encoding the zinc finger DNA binding domain alone (Δ253EKLF), a region shown previously to be sufficient for chromatin remodeling in vitro, demonstrate erythroid and megakaryocytic phenotypes that resemble those observed in EKLF-null hematopoietic progenitors. Taken together, our results suggest strongly that the unique EKLFCR domain is necessary and sufficient to modulate the chromatin-specific roles of EKLF at erythroid- and megakaryocytic-specific loci in definitive hematopoietic cells in vivo.