bluuebunny/crossref_metadata_2025_split huggingface dataset:
Natural History of Transfusion-Independent Non-Severe Aplastic Anemia in Children.
Authors: Nishio Nobuhiro , Yagasaki Hiroshi , Takahashi Yoshiyuki , Hama Asahito , Muramatsu Hideki , Kojima Seiji 
Published: October 2019

Abstract
The known clinical courses of non-severe aplastic anemia (NSAA) in children are spontaneous resolution, persistent NSAA, or progression to severe aplastic anemia (SAA). A few published reports have indicated the outcome of transfusion-independent NSAA. We performed a retrospective analysis of transfusion-independent NSAA to evaluate the incidence and time of progression to transfusion-dependent NSAA or SAA. We reviewed the records of 70 children with acquired aplastic anemia referred to Nagoya University Hospital between September, 1986 and August, 2006. Transfusion-independent NSAA was defined as hypocellular marrow and at least 2 cytopenias that persisted for several weeks, without meeting the criteria for SAA (neutrophil count less than 500/μl, a platelet count less than 20,000/μl, and an absolute reticulocyte count less than 20,000/μl). We included only children (aged less than 18 years) with transfusion-independent NSAA at diagnosis who were treated with supportive care alone. The median ages of the 23 patients who met the criteria were 7.7 years (range, 3.0–15.2 y) and 15 were female. The etiology of the aplastic anemia was unknown in all patients. At diagnosis, median neutrophil, platelet and reticulocyte counts were 1,040/μl (range, 380–3,200/μl), 41,000/μl (range, 20,000–126,000/μl) and 63,000/μl (range, 10,000–120,000/μl), respectively. Of 22 patients for whom chromosomal data were available, none had chromosomal abnormalities at diagnosis. Patients were followed for a median of 89 months (range, 11–198 months). Transfusion-independent NSAA persisted in 10 (43%) patients at a median of 30 months (range, 11–117 months) after diagnosis. On the other hand, 13 (57%) progressed to transfusion-dependent NSAA or SAA at a median of 53 months (range, 9–175 months) after diagnosis and required some treatment. Of these 13 patients, 4 received androgens, 2 received high-dose methylprednisolone, 1 received immunosuppressive therapy (IST) with anti-thymocyte globulin and cyclosporine, and 4 underwent hematopoietic stem cell transplantation (HSCT) from HLA-matched siblings. The Kaplan-Meier estimate for progression-free survival was 64%±11% at 5 years and 25%±12% at 10 years, respectively. None of the patients treated with supportive care alone had hematological improvement. None developed paroxysmal nocturnal hemoglobinuria or myelodysplastic syndrome, and all patients are currently alive. In conclusion, NSAA did not spontaneously resolve in any of the 23 patients and the incidence of disease progression was high. Whether early intervention with IST or supportive care alone is preferable should be compared.
The Protective Role of Protein C in Endotoxin-Induced Shock.
Authors: Lay Angelina J. , Donahue Deborah , Tsai Meng-Ju , Castellino Francis J. 
Published: October 2019

Abstract
The serine protease, activated Protein C (aPC) plays an important role in the maintenance of vascular hemostasis. Besides its ability to regulate coagulation and fibrinolysis, aPC also possesses antiinflammatory and antiapoptotic properties. Severe depletion of plasma PC and aPC contributes to sepsis pathogenesis. Although treatment with recombinant aPC benefits a subset of patients with severe sepsis, mechanisms by which aPC improves survival in these patients remain largely unknown. Using mice genetically predisposed to a severe PC deficiency, we initiated studies to further elucidate the mechanistic relationships between very low endogenous PC levels and inflammatory disease pathogenesis. Here, we show for the first time, that novel genetic dosing of PC strongly correlates with survival outcome following endotoxin (LPS) challenge in mice. Our findings provide evidence that very low endogenous levels of PC predispose mice to early onset of disseminated intravascular coagulation, thrombocytopenia, hypotensive shock, organ damage, and reduced survival after LPS challenge. Furthermore, an excessive inflammatory response is observed in very low-PC mice, but is greatly reduced in WT cohorts. Administration of recombinant human aPC to low-PC mice significantly prevents endotoxin-induced hypotensive shock and prolongs survival. This study highlights the importance of host endogenous levels of PC in predicting survival outcome following a severe acute inflammatory challenge.
Antithrombin Deficiency in Pregnancy.
Authors: Andrew Retter J. , Beverley Hunt J. 
Published: October 2019

Abstract
Background: During pregnancy untreated antithrombin deficiency is associated with up to a 50% risk of venous thromboembolism (VTE) and a relative risk of pregnancy loss of 2.1 with a 5-fold increase in stillbirths. Thus thromboprophylaxis is widely used, but little data is available to select type, dose & duration of anticoagulation.
Method: We performed a retrospective, single centre observational study of our antithrombin deficient pregnancies since 1996.
Results: There were 9 pregnancies in 8 women; median age at conception 33 (age-range 19–37). They separated into 3 groups (1) 4 asymptomatic patients diagnosed on family screening. They received unmonitored enoxaparin 40mg until 16 weeks then 40mg BD. (2) 2 with previous VTE, received intermediate dose enoxaparin (1mg/kg), increased to BD at 16 weeks. Monitoring was done to maintain an anti-Xa trough of <0.12 iu/ml and peak <0.8iu.ml. (3) 2 referred after presenting with VTE in pregnancy. They received enoxaparin 1mg/kg BD and the same monitoring These included a known antithrombin deficient woman, referred in her second pregnancy at 26weeks gestation with premature rupture of the membranes and an iliofemoral deep vein thrombosis which developed on enoxaparin 60mg OD. Enoxaparin was increased to 1mg/kg BD and an IVC filter inserted. Despite the filter however she had a pulmonary embolism. The filter was removed after Caesarean section at 31 weeks. Two had sagittal sinus thromboses in the first trimester associated with severe hyperemesis requiring IV fluids. One was our only thromboprophylaxis failure, receiving enoxaparin 40mg OD, she weighed 80Kg. The second presented at 11weeks gestation. She was intolerant of self injecting and so switched to warfarin at 15 weeks until 35 weeks as did one other mother. All mothers had close feto-maternal monitoring with uterine artery Doppler at 24 weeks if possible and then monthly growth scans thereafter.
Delivery: Thromboprophylaxis was stopped at labour initiation or 12hrs prior to Caesarean section (3 women) and 50iu/kg of antithrombin concentrate was given. Anticoagulation was restarted 24hrs after delivery. Six weeks enoxaparin post-partum thromboprophylaxis was given or the women converted back to warfarin. Estimated blood loss at delivery was a median of 200ml (range 200–500ml), no transfusions were required. There were no post partum VTEs. Nine births occurred at a median gestation of 38weeks (range 31–41), median birth weight 3045g (range 1420–4120g). One child has West’s syndrome.
Conclusion: This is the largest case series on the management of antithrombin deficiency in pregnancy. The combined use of enoxaparin in pregnancy and post partum combined with antithrombin concentrate during labour appears to improve pregnancy outcome and reduce the rate of VTE. Larger studies are required to confirm this finding.
Bcl-xS Is an Important Pro-Apoptotic Bcl-2 Family Member in Chronic Lymphocytic Leukaemia and Its Expression Is Regulated by SAM68, hnRNP-F and hnRNP-H.
Authors: Willimott Shaun , Wagner Simon D. 
Published: October 2019

Abstract
We have previously shown that the pro-apoptotic Bcl-x isoform Bcl-xS is induced in chronic lymphocytic leukaemia cells during spontaneous and drug induced apoptosis to a greater extent than other pro-apoptotic Bcl-2 family members including Bim, Puma and Noxa. Puma, Noxa and also Bid were induced strongly by stimulation with CD40 ligand, in culture conditions causing CLL cell proliferation and survival, and associated with strong induction of the pro-survival Bcl-x isoform, Bcl-xL. Immunohistochemistry demonstrates Bcl-xL expression in lymph node proliferation centres. We, therefore, conclude that splicing of Bcl-x mRNA is important for regulation of CLL survival such that Bcl-xL is required during proliferation in the lymph node or bone marrow microenvironment, but Bcl-xS is induced during apoptosis. Splicing of Bcl-x mRNA in HeLa cells is known to be controlled by RNA binding proteins including SAM68, heterogeneous ribonucleoprotein-F (hnRNP-F) and hnRNP-H, but expression and regulation of these molecules has not been investigated in CLL. We have demonstrated that expression of hnRNP-F and hnRNP-H decline in association with an increase in Bcl-xS expression. Conversely expression of the ribonucleoproteins is maintained (relative to expression in freshly isolated peripheral blood CLL cells) or increases on CD40 stimulation. SAM68 showed a similar pattern of expression with increased expression on CD40 stimulation and decreased under conditions of apoptosis. Phosphorylation of SAM68 can control the RNA binding activity of this molecule. We immunoprecipitated SAM68 from freshly isolated CLL cells and cells that had been cultured with CD40 ligand (to promote proliferation) or on tissue culture plastic (to promote apoptosis) for 2 days. Westerns of the immunoprecipitated protein were probed with an anti-phosphotyrosine antibody. We found variable patterns: phosphorylation was uniformly present in cells cultured on plastic (4/4 cases) but was only detectable in freshly isolated cells in 1 case and on CD40 stimulation in 2 cases. We conclude that the RNA binding proteins hnRNP-F, hnRNP-H and SAM68 have important roles in controlling Bcl-x splicing in CLL cells, and that their effects are largely due to variation in expression level rather than phosphorylation. Strategies to bias production of pro-apoptotic Bcl-xS using siRNA either directed against Bcl-xL mRNA or against RNA binding proteins may be therapeutically useful.
High Level EBV Persistence in the IgM+ IgD+ CD27+ B Cell Subset in Patients with X-Linked Lymphoproliferative Disease Lacking Isotype-Switched Memory B Cells.
Authors: Chaganti Sridhar , Ma Cindy , Bell Andrew , Croom-Carter Debbie , Hislop Andrew , Tangye Stuart , Rickinson Alan 
Published: October 2019

Abstract
Epstein-Barr virus (EBV) infects >90% of population world wide and, in healthy virus carriers, establishes life long persistence in the immunoglobulin (Ig)Dneg, CD27+ (“class-switched”) memory B cell compartment normally produced by antigen stimulation and transit through germinal centres. Patients with the X-linked lymphoproliferative disease (XLP) cannot make such class-switched memory B cells due to an inherited mutation in the slam-associated protein (SAP) gene involved in the maturation of antibody responses. Interestingly, XLP patients are highly susceptible to severe primary EBV infection and develop a fulminant infectious mononucleosis (IM) which is often fatal and where the symptoms progress to resemble those of a different (non-familial) disease, EBV-associated haemophagocytosis syndrome (EBV-AHS), caused by virus entry into the NK or T cell system. Some XLP patients survive their primary infection, but the nature of EBV carriage in these individuals (lacking conventional memory B cells) remains unresolved. To investigate this further we obtained blood samples from 8 such XLP patients. EBV load in total peripheral blood mononuclear cells (PBMC), determined by quantitative PCR, occupied a broad range but on average was 2 to 3-fold fold higher than that of healthy controls. The virus was concentrated within the B cell (CD19+) compartment, as in healthy carriers, and not within T or NK cells, as typically seen in EBV-AHS. We then confirmed that these XLP patients indeed lacked conventional class-switched memory B cells but did carry a small population of IgM+, IgD+, CD27+ (“non-switched”) memory cells; however their circulating B cell pool was dominated by naïve (IgM+, IgD+, CD27neg) cells and by expanded numbers of immature “transitional” (CD10+ CD27neg) cells. In each of 4 cases studied by cell sorting, EBV was concentrated in this small subset of “non-switched” memory B cells. To see if the high virus load detected in these patients indicated true virus persistence as opposed to recent or recurrent infection, serial samples obtained over a 3 year period from two XLP patients were assayed. Virus load was stable and, in one case with the highest load, screening with markers of virus polymorphism detected the same resident strain over time. Our results in XLP patients make it clear that EBV can persist in the absence of a conventional class-switched memory B cell compartment. Instead, the virus is sequestered in a small population of “non-class-switched memory” cells with Ig gene mutations. The origin of such cells, which are also detectable in the blood of normal donors, is uncertain; however their existence in XLP patients suggests that such cells arise independently of germinal centre activity and hence that EBV may be able to colonise its host without exploiting germinal centre transit.
Population Pharmacokinetics (PK) of Oral Posaconazole (POS) for Antifungal Prophylaxis in Neutropenic Patients with Hematologic Malignancies.
Authors: AbuTarif Malaz A. , Krishna Gopal , Statkevich Paul 
Published: October 2019

Abstract
In a recent large, randomized, multicenter study in neutropenic patients undergoing remission-induction chemotherapy for acute myelogenous leukemia or myelodysplastic syndrome, prophylaxis with POS resulted in significantly fewer invasive fungal infections (IFIs) (2% vs 8%, P<0.001) and longer survival (P=0.04) vs fluconazole or itraconazole (Cornely et al New Engl J Med2007;356:348–59). Using data from this study, the relationship between demographic/baseline covariates and apparent volume of distribution (V/F) of POS was evaluated in a population modeling approach. In the full study, 304 patients were administered POS (200 mg tid) and 298 patients received fluconazole (400 mg qd) or itraconazole (200 mg bid) for up to 12 weeks. For the population PK analysis, data from 215 POS patients were analyzed using nonlinear mixed-effect modeling. The effect on POS V/F of IFI occurrence, demographic characteristics, and baseline variables (occurring at or before day 7) including mucositis, neutropenia, vomiting, diarrhea, proton pump inhibitor (PPI) or H2-receptor antagonist usage, bilirubin ≥2 × upper limit of normal (ULN) and gamma-glutamyl transferase (GGT) ≥2 × ULN was examined. A 2-stage “stepwise forward addition” followed by a “stepwise backward elimination” approach was used to build the covariate model. The model was validated by bootstrapping data with 85% and 50% partial sampling. POS V/F, and thus exposure, was statistically associated with diarrhea, PPI intake, bilirubin ≥2 × ULN, GGT ≥2 × ULN, and race (white vs nonwhite). None of these covariates predominated in patients with proven or probable IFI (n=7). Validation demonstrated the robustness of this model. A significant relationship was found between POS V/F (or exposure) and the following covariates: diarrhea, PPI intake, bilirubin ≥2 × ULN, GGT ≥2 × ULN, and race. However, the influence of these covariates on POS exposure is unlikely to be clinically relevant as none of the covariates predominated in the patients who developed IFI, and plasma POS levels were attained that were adequate for successful prophylaxis against IFIs. Thus, no dosage adjustments are recommended based on these covariates.
Inhibition of the TGF-β Receptor I Can Stimulate Hematopoiesis in Primary Myelodysplastic Syndrome Progenitors as Well as in TGF-β Driven Transgenic Mouse Model of Bone Marrow Failure.
Authors: Zhou L. , Nguyen A. , Pahanish P. , Hayman J. , Gundabolu K. , Chubak A. , Parmar S. , Garry D. , Wickrema A. , Navas T. , Higgins L. , Friedman E. , List A. , Bitzer M. , Verma A. 
Published: October 2019

Abstract
Myelodysplastic syndromes (MDS) are characterized by ineffective hematopoiesis that leads to peripheral cytopenias. TGF-β is a myelosuppressive cytokine that has been indirectly linked to the pathogenesis of some subsets of MDS and acute leukemias. We demonstrate by immunohistochemistry that smad2, a component of the TGF-β signaling pathway, is constitutively activated in MDS bone marrows. This activation was seen in both low and high grade cases of MDS when compared to anemic controls and provides the first direct evidence of activation of TGF-β signaling in MDS. To demonstrate the functional role of TGF-β signaling in MDS, a GFP expressing lentiviral siRNA construct was designed to knockdown TGF receptor I (TBRI) expression. Stable expression of this lentivirus in a variety of hematopoietic cells resulted in >70% inhibition of TBRI mRNA by qPCR. Lentiviral siRNA Knockdown of TBRI in primary CD34+ cells resulted in an increase in GFP+ erythroid colony formation, indicating an inhibitory role of TGF-β in human hematopoiesis. To further test the efficacy of TBRI inhibition in stimulating hematopoiesis, we used a specific and potent inhibitor of TBRI (ALK5) kinase, SD-208. SD-208 was effectively able to inhibit TGF-β mediated activation of smad-2 and potently inhibit TGF-β mediated gene expression in bone marrow stromal cells. SD-208 treatment also led to reversal of TGF-β induced inhibition of primary CD34 cell proliferation and both erythroid and myeloid colony formation in short term assays. Most importantly, treatment with SD-208 led to significant dose dependent increases in erythroid and myeloid colony formation from primary MDS bone marrow derived hematopoietic progenitors. Furthermore, we tested the efficacy and specificity of SD-208 in a TGF-overexpressing transgenic mouse. This mouse constitutively expresses TGF-β through an albumin promoter, develops progressive anemia and dysplasia and thus serves as a novel model of human bone marrow failure. Treatment with SD-208 by oral lavage at 30mg/kg/d for two weeks led to significant increases in both myeloid and erythroid colony formation from murine bone marrows and led to a trend of increasing hemoglobin and hematocrit. Taken together, these studies demonstrate a role of TBRI inhibition in stimulating hematopoiesis in human bone marrow failure and should lead to future studies with SD-208 and other inhibitors of TGF signaling pathways in MDS.
Lentiviral Insertion Site Analysis of Corrected Mouse Model of β Thalassemia Following Lentiviral Transduction of Embryonic Stem Cells.
Authors: Yang Rui , McConnell Sean C. , Huo Yongliang , Ulrey Clayton , Liu Shan-Run , Zhang Ting-Ting , Ryan Thomas M. 
Published: October 2019

Abstract
The delivery of therapeutic transgenes via lentiviral transduction of stem cells holds great promise for future cell based therapies for inherited genetic disorders. The regulation of any transgene integrated into patient derived embryonic stem (ES) cells (generated via nuclear transfer or reprogramming) needs to be studied in great detail prior to their differentiation and transplantation back into the patient. Because genetically manipulated ES cells can be clonally isolated, expanded to great numbers in an undifferentiated state, and differentiated to specific cell types, they can be carefully tested in vitro prior to their reintroduction back into patients. ES cells derived from a mouse model of β thalassemia were transduced with a recombinant, self inactivating (SIN) lentiviral vector containing a 2.3 kb human β-globin gene and a 3.2 kb Locus Control Region composed of regulatory elements; HS2, HS3, and HS4. Clonal populations of transfected β thalassemic ES cells were isolated, archived, and utilized to produce genetically identical offspring by injection into tetraploid blastocysts or eight-cell embryos. Real-Time QPCR expression analyses demonstrated high levels of human β-globin gene expression in the peripheral blood of five of the six lines produced. The average expression level per transgene copy number ranged from 13% to 62% of endogenous mouse α-globin levels. Peripheral blood hemolysates analyzed by HPLC confirmed the high level production of human β-globin chains. Measurement of the red blood indices in these five lines showed that the anemia was corrected. In order to examine the influence of chromosomal position effects on individual transgene expression, the lentiviral insertion sites were mapped by linear amplification (LAM) PCR. Cloned mice from the six lines were bred in order to segregate each of their individual lentiviral transgenes. Offspring containing single insertion sites were analyzed for human β-globin transgene expression and synthesis by Real-Time QPCR of blood RNA and by HPLC of hemolysates, respectively. These analyses demonstrate that there is a preference for provirus integration in ES cells; all transgene insertions are not expressed equally; lentiviral transduction of a human β-globin gene into ES cells can cure a severe hemoglobinopathy; and that the delivery of therapeutic transgenes via lentiviral transduction of ES cells did not harm the developmental potential of the cells to produce an entire mouse.
STAT5 Signaling Is Associated with the Outcome of Follicular and Mantle Cell Lymphoma Patients Treated with Immunochemotherapy.
Authors: Leppä Sirpa , Taskinen Minna , Sommarhem Antti , Elonen Erkki , Nykter Matti , Monni Outi , Timonen Tuomo , Hautaniemi Sampsa , Karjalainen-Lindsberg Marja-Liisa , Meri Seppo 
Published: October 2019

Abstract
Addition of rituximab (R) to chemotherapy (immunochemotherapy) has significantly improved the outcome of B-cell lymphoma patients. However, responses are still unpredictable due to heterogeneous nature of these diseases. The aim of this study was to determine if differences in gene expression in follicular lymphoma (FL) and mantle cell lymphoma (MCL) tissue correlate with outcome in response to immunochemotherapy. 24 FL and 15 MCL patients were classified into complete (CR) or other responders after immunochemotherapy, and genes capable of separating the groups were identified using oligonucleotide-based microarray and supervised learning technique. One of the transcripts associated with increased CR rates in FL was signal transducer and activator of transcription 5a (STAT5a), a transcription factor and a mediator of cellular responses after cytokine stimulation. According to Kaplan Meier estimates, high STAT5a mRNA levels were associated with better progression free survival (PFS). Immunohistochemical analyses of FL samples demonstrated that predominant STAT5a activity localized to resident T-lymphocytes but only 20% of the samples were STAT5a positive. In a validation set of 79 FL patients, a better PFS was observed among patients with high STAT5a protein expression. In contrast, high STAT5a mRNA and protein levels were associated with poor outcome in MCL patients. In the MCL tissue, STAT5a expression localized to nonmalignant T-lymphocytes but also to tumor cells. In comparison to FL, T-lymphocytes were more uniformly positive for STAT5a. In sum, the data show that nonmalignant tumor cells have a profound prognostic impact both in FL and MCL, but also suggest that depending on the lymphoma subtype, the activity of a certain signaling pathway can have either positive or negative consequences on the survival of immunochemotherapy-treated lymphoma patients.