bluuebunny/crossref_metadata_2025_split huggingface dataset:
Erythropoietin Signaling and Response in Vascular Smooth Muscle Cells.
Authors: Beleslin-Cokic Bojana B. , Cokic Vladan P. , Gojkovic-Bukarica Ljiljana , Noguchi Constance Tom 
Published: April 2020

Abstract
Erythopoietin (EPO) and erythropoietin receptor (EPOR) regulate survival, proliferation, differentiation and viability of erythroid progenitor cells. Beyond erythropoiesis, we have observed that human vascular endothelial cells respond to EPO stimulation by inducing EPOR and endothelial nitric oxide synthase (eNOS) expression, increasing NO production and cGMP, particularly under low oxygen tension. In this study, we investigate the response of vascular smooth muscle (VSM) to EPO stimulation and the contribution of blood vessel to relaxation/contraction. We found that VSM cells express EPOR and that treatment with EPO (5 U/ml) at reduced oxygen tension increased EPOR mRNA by 2 fold. This increased EPO responsiveness at low oxygen tension is accompanied by increased cell proliferation at 2% O2 more than 2 fold. Unlike endothelial cells, EPO did not induce eNOS or NO production in VSM cells. PI-3 kinase was involved in EPO stimulation with no change in MAP kinase. In an isolated blood vessel model system, we checked EPO responsiveness. EPO produced contractions (0.32 ± 0.3 g) of rat renal artery, and pretreatment with LY294002 (10 mM), an inhibitor of PI-3 kinase, statistically significantly inhibited this contraction (58.7 ± 7%). This response was also observed with the endothelium layer removed. In preparations of human internal mammary artery (HIMA) and human saphenous vein (HSV) from patients undergoing coronary artery bypass, EPO did not affect basal vascular tone of HIMA and HSV with the endothelium layer removed, but EPO (5 U/ml) potentated noradrenalin-induced contraction by up to 2 fold in HSV and HIMA. Also, pretreatment with EPO significantly increase angiotensin-evoked contractions of these blood vessels (50 ± 8%, 113 ± 17%, respectively P &lt; 0.01), suggesting that EPO has synergistic effects on angiotensin or noradrenalin-induced [Ca2+] mobilization, particularly on intracellular Ca2+ release. These data suggest that EPO stimulation of vascular smooth muscle may act to modulate or balance the vasodilatory effects of increased NO production by EPO stimulated endothelium. Under oxygen stress vascular smooth muscle can respond to EPO by proliferation and PI-3 kinase activation as a protective effect and in conjunction with Ca2+ release likely contributes to the overall vascular response.
Non-Integrating Gene Transfer To Redirect Specificity of Lymphocytes towards Pediatric CD19+ Malignancies
Authors: Yuen Carrie H. , Andersson Helen , Olivares Simon , Biswal Lisa , Raphael Robert , Killian Tom , Shpall Elizabeth J. , Lee Dean A. , Champlin Richard , Cooper Laurence 
Published: April 2020

Abstract
Pediatric B-lineage malignancies, such as acute lymphoblastic leukemia (B-ALL), generally have a poor prognosis upon relapse of tumor cells exhibiting resistance to conventional chemotherapy. In the absence of new commercially-available therapies for refractory pediatric cancers, it is up to pediatric oncologists to develop and apply new therapeutics. One candidate targeted therapy for B-lineage leukemia/lymphoma, is adoptive transfer of autologous T cells genetically modified to express a chimeric antigen receptor (CAR) and rendered specific for CD19. However, the broad adoption of T-cell gene therapy for children is limited by the expense to produce and release expression vectors and cell products and the risk of insertional mutagenesis. Therefore, we are developing an approach to express the CAR without integration of transgene and therefore without causing genotoxicity. Here we demonstrate that non-viral gene transfer of vector-free in vitro-transcribed messenger ribonucleic acid (mRNA) can efficiently introduce CAR into T cells. We demonstrate, using a commercially available electroporator, that codon-optimized mRNA coding for CD19-specific CAR is expressed on primary human T cells (Figure 1A) that redirects the specificity of CD19+ tumors (Figure 1B). When scaled-up for clinical trials, the electro-transfer of one or more mRNA species into T cells has the advantage of (i) ex vivo manipulation of T cells with preservation of homing receptors for lymph nodes and bone marrow (sites of leukemia/lymphoma), (ii) availability of clinically-meaningful numbers of genetically manipulated T cells within a day of apheresis, (iii) high levels of transgene expression, (iv) reduced cost (preparation of clinical-grade plasmid and mRNA compared with recombinant virus), and (iv) safety (no insertional mutagenesis). The relative ease of gene/mRNA transfer coupled with minimal manipulation of large numbers of T cells allows investigators to give patients multiple doses using existing infrastructure such as already widely exists for processing hematopoietic stem cells and blood banking. We are preparing a multi-institution clinical trial in which pediatric patients with refractory B-ALL receive personalized medication, prepared at each site, in the form of recursive infusions of CAR+ T cells repetitively electroporated with mRNA. This application of transient transfection to adoptive immunotherapy is expected to be of wide appeal for pediatric and adult patients alike with tumors that can be targeted by CAR.
Figure: (A) Expression of CAR in primary human T cells. (B) Specific lysis of CD19+ tumor target by genetically manipulated T cells expressing CAR mRNA Figure:. (A) Expression of CAR in primary human T cells. (B) Specific lysis of CD19+ tumor target by genetically manipulated T cells expressing CAR mRNA
A Heat-Inducible Gene Expression System for a Dominant Negative Version of a TGF-β-Receptor Reveals Antitumor Activity in a Rodent Tumor Model
Authors: Noyan Fatih , Dewey Ricardo A. , Kreipe Hans H. , Fruehauf Joerg , Klein Christoph 
Published: April 2020

Abstract
In an attempt to design novel therapeutic strategies against solid tumors, we hypothesized that spatial and temporal control of transgene expression may allow for the delivery of immunomodulatory molecules at the site of tumor. Here, we assess the potential of the HSP70b promotor to control lentivirus-mediated gene expression in tumor-infiltrating hematopoietic cells. We generated a series of lentiviral vectors and characterized transgene expression in vitro and in vivo. Up to 20fold induction of GFP expression was seen when lentivirus-transduced cells were heated to 43°C in vitro. Only minimal leakiness was seen at 37°C. Next, we analyzed HSP-driven GFP expression in hematopoietic cells upon transplantation of lentivirus-transduced hematopoietic stem cells, using constitutively active promotors and empty vectors as controls. Mice were inoculated with the glioma cell line GL261 and GFP expression was assessed in various organs as well as in tumor-infiltrating hematopoietic cells. Interestingly, HSP70-driven transgene expression was 10–15fold higher in tumor-infiltrating cells when compared to spleen cells, suggesting that the HSP70b promoter was active in the tumor microenvironment. To assess whether confined expression of a dominant negative TGF-β-receptor within the tumor microenvironment would induce anti-tumor immunity, we transplanted mice with hematopoietic stem cells transduced with TGF-β-DNR and injected GL261 cells subcutaneously. The tumor volume was decreased up to 40 fold in mice expressing HSP70b-TGF-β-DNR, comparable to mice transplanted with hematopoietic stem cells that constitutively express TGF-β-DNR. Functional studies suggest a role for perforin, IFN-α and Granzyme B in this TNF-β-DNR-dependent immunomodulatory circuit. In summary, HSP70b-mediated transgene expression in hematopoietic cells may provide a novel experimental strategy for controlled release of immunmodulatory molecules to induce anti-tumor therapies.
Influence of Polymorphisms of Pro-Inflammatory (IL-12, TNFa and LTa) and Anti-Inflammatory Factors (IL-10 and CTLA4) in Autoimmune Hemolytic Anemia.
Authors: Figueiredo Maria S. , D’Abronzo Leandro S. , Oliveira Melca M. , Dreyfuss Juliana L. , Bordin Jose Orlando 
Published: April 2020

Abstract
The Autoimmune Hemolytic Anemia (AIHA) is caused by the destruction of antibody-coated red blood cells, but mechanisms that initiate the production of autoantibodies remains unclear. It had been suggest that decreased production of Th1-type cytokines and production of autoantibodies in AIHA can be secondary to the imbalance between anti- and pro-inflammatory factors. Moreover, the presence of single nucleotide polymorphisms (SNPs) showed association with different production of immunoregulatory factors which may modulate the disease expression in AIHA.
OBJECTIVES: To determine the importance of SNPs of pro- and anti-inflammatory factors in the development of AIHA.
PATIENTS: We studied 17 patients with AIHA who has been followed in the Hematology and Blood Transfusion Unit at the Federal University of Sao Paulo (UNIFESP/EPM), Brazil. The control group was composed by 40 healthy volunteer blood donors.
METHODS: After DNA extraction from peripheral blood samples, the frequency of the SNPs was determinate by PCR-RFLP in patients and healthy individuals. The following SNPs were analyzed: Interleukin 12: IL-12 1188 (A/C), Tumor Necrosis Factor alpha: TNFa-308 (G/A), and Lymphotoxin alpha: Lta +252 (A/G); Interleukin 10: IL-10-592 (C/A), and Cytotoxic T-lymphocyte associated protein 4: CTLA4 exon 1 49 (A/G).
RESULTS: The patient group was composed predominantly by female individuals (14 or 82%) and the median of age was 56 years old (18 to 76 years). The frequency observed for each allele studied in the patient group was: allele A of IL-12 = 0.82; allele G of FNTa = 0.85; allele A of Lta = 0.68; allele C of IL-10 = 0.82; allele G of CTLA4 = 0.59. No differences in allele frequency were found between patient and control groups.
CONCLUSIONS: Our results suggest that these polymorphisms appear not to contribute for the development of the AIHA.
Screening of Pathogenicity-Related Genes in Bone Marrow CD4+ T Cells of Aplastic Anemia Patients.
Authors: Zheng Miao , Liu Wenli , Sun Hanying , Zhou Jianfeng , Zhang Yicheng 
Published: April 2020

Abstract
Recent clinical and experimental data have proved that CD4+ T cells play a crucial role in the pathogenesis of aplastic anemia (AA). However, the mechanisms how CD4+ T cells in AA over-proliferate, activate, infiltrate bone marrow and damage hematopoietic cells have largely remained unknown. Therefore we adopted the suppressive subtractive hybridization (SSH) method so as to screen differentially expressed genes of bone marrow CD4+ T cells between AA patients and normal donors, which may hopefully help clear up the mystery of the pathogenesis of AA. Firstly, we selected a pair of subjects: a typical first-visit AA patient and a healthy donor of the same age and sex. Their bone marrow (BM) mononuclear cells were isolated using lymphocyte separating medium by density gradient centrifugation. After a 24-hour expansion culture, CD4+ T cells were separated with magnetic bead sorting technique. Secondly, with CD4+ T cells of the patient as “tester” and those of the normal donor as “driver”, a cDNA library was established by the SSH method. 110 clones were detected by means of PCR in the established subtractive library, which contained an inserted fragment with 200∼700bp respectively, and the positive detected rate was up to 88%. Then 10 of the resulting subtracted cDNA clones were randomly selected for a respective DNA sequencing and honologic analysis. Among 10 sequenced clones, there were 8 known genes including 2 repeated genes. Lastly, we collected BM samples of 20 AA patients and 20 healthy donors so as to validate the expression levels of differentially expressed genes from SSH library with semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR). We found, compared with normal donors, there were 6 genes among these 8 genes from SSH library up-expressed in BM CD4+ T cells of patients with AA. These genes were transducin (beta) -like 1X-linked receptor 1 (TBL1XR1) (NM024665); ATP-binding cassette, sub-family B (MDR/TAP), member 6 (NM005689); ARP2 actin-related protein 2 homolog (ACTR 2) (NM005722); zinc finger protein 561 (ZNF561) (NM152289); NADH dehydrogenase (ubiquinone) 1 beta subcomplex, 6 (NM002493); eukaryotic translation initiation factor 3, subunit 5(NM003754). These results indicate that some genes are found able to regulate functions of BM CD4+ T cells in AA, concerned with protein synthesis, biology oxidation, signal transduction and cell migration, etc. Therefore, screening and cloning these genes are helpful to elucidate the possible mechanisms of AA.
The Effects of Tcf-1 Gene Silence on Marrow CD4+ T Cells from Aplastic Anemia Patients.
Authors: Zheng Miao , Liu Wenli , Sun Hanying , Zhou Jianfeng , Zhang Yicheng 
Published: April 2020

Abstract
As T-cell special expression gene, T cell factor 1 (Tcf-1) has an HMG-BOX domain be cognized as a transcription factor. We have found from our previous studies that Tcf-1 mRNA expression level in bone marrow (BM) CD4+ T cells from aplastic anemia (AA) patients is markedly higher than that of normal people. It is presumed that up-expression of Tcf-1 gene can be closely related to abnormal functions of BM CD4+ T cells from AA patients. Therefore in the current study, we discussed the effects of Tcf-1 gene silence on the BM CD4+ T cells from AA patients. Three shRNA expression vectors psiRNA1, psiRNA2 and psiRNA3 were constructed with psiRNA-hH1neo G2 plasmid. They were respectively transfected into Jurkat cells in which there was rather high expression level of Tcf-1 gene, then Tcf-1 mRNA of Jurkat cells was assayed by semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR). All three plasmid-derived siRNAs could effectively reduce the expression level of Tcf-1 mRNA in transfected Jurkat cells, but the plasmid-derived siRNA psiRNA2 was found to be the most effective inhibitor of Tcf-1 expression. Next, the selected psiRNA2 were transfected into the BM CD4+ T cells from six AA patients. It was found that the expression level of Tcf-1 mRNA in BM CD4+ T cells from these patients was reduced by 45.0%. At the same time, the expression levels of c-myc and CD44 mRNA in BM CD4+ T cells from these AA patients were also decreased to different extents, 47.7% and 53.7%, respectively. These results indicate that Tcf-1 gene can regulate downstream target genes such as c-myc and CD44 to participate in over-proliferation and activation of BM CD4+ T cells from AA patients. It is likely worthwhile to carry out the in-depth study against Tcf-1 in AA therapy.
Anemia an Old Player or a Cut-Throat in Patients with Coronary Artery Disease or Chronic Heart Failure.
Authors: Corinaldesi Giorgio , Corinaldesi Christian 
Published: April 2020

Abstract
Anemia is associated with an increased number of adverse cardiovascular events (CVD) in particular with coronary artery disease (CAD), and chronic heart failure (CHF), and it is also correlated with gender, aging, renal insufficiency, low BMI. Anemia involves inflammatory cytokines (C-reactive protein, IL-6, MCP-1, TNF-alfa), it reduces marrow response to erythropoietin (EPO) and heme-oxygenases-1(HO-1), it also reduces red cells life span and it may impairs reuse of iron, it mostly reduces, the peak VO-2 (peak aerobic power). The latter appears to be an independent factor that may be associated with an adverse outcome, in fact, for a reduction of one gram of heamoglobin (Hb) the risk of morbility and mortality increase respectively by 32% and 18%. The aim of the study was to determine the clinical implication of anemia in patients with CHF or CAD; we have studied 48 patients (32 male, 16 female) with CHF, and 52 patients (34 male, 18 female) with CAD, with a range of Hb concentration included between 9.4gr/dl and 12.6gr/dl. We have evaluated moreover the tolerance to exercise on a treadmill and six minute walk distance (210+/− 32 m in CAD), (180+/− 28 m in HF), the presence of rest dyspnea, the presence of supraventricular or ventricular arrhythmias (atrial/or ventricular premature beats, sinus tachycardia, or ventricular tachycardia, atrial fibrillation); lower levels of Hb, Fe, TIBC correlate with a greater tendency to develop ventricular arrhytmias instead of supraventricular arrhytmias. Anaemia management included erythropoietin stimulating protein, blood transfusion; we have used darbopoietin 50 mcg every week, and this treatment is associated to a significant improvement in functional class and cardiac and renal function. Epo has a strong cardioprotective effect: reducing left ventricular hypertrophy, infarct size, a higher beta natriuretic peptide level, apoptotic cell death, increase FE and capillary vessels; we have remarked a longer endurance time of exercise testing a greater distance walked (282+/− 64 m in CAD), (248+/− 32m in CHF), a significant increased in the peak oxygen consumption VO2 from 12.6+/− 2.4 to 16.2+/− 2.8 mL/Kg x min. in CAD, 9.8+/− 2.0 to 13.2 +/− 3.4 mL/Kg x min. in CHF. Our data also confirm the link between an increased tendency to develop CVD and a decreased level of Hb.
RISK FACTORS CAD CHF Sex (M/F) 32/16 34/18 Age (Years) 62+/− 6 66+/− 4 Hb (g/dl) 10.8+/− 1.8 10.6+/− 1.2 MCV (fl) 82.6+/− 4.4 77+/− 4.2 Iron (mg/dl) (Fe) 42.8+/− 10.2 36+/− 9.4 Total Iron Binding Capacity (TIBC) 316+/− 68.8 280+/− 62.8
Parvovirus B19 Detection by PCR in Patients with Chronic Renal Failure and Resistance to Erythropoietin: An Interim Report.
Authors: Franco Hector , Escoboza Jose R. Borbolla , Scott Sean P. , Villela Luis M. , Valdez Alejandro , Aviña Ana Elena , Benavides Guillermo 
Published: April 2020

Abstract
Introduction: The human parvovirus B19 (pvB19) is well known as a cause of chronic anemia and red cell aplasia, this due to the high tropism for the erythroid progenitor cells where it replicates. Some reports have suggested that the infection of this virus is related to an erythropoietin resistance in patients with chronic anemia and kidney failure. The most frequent causes of erythropoietin resistance are iron deficiency, chronic inflammation process, vitamin deficiency, hyperparathyroidism and malignancies among others. The aim of this study was to detect the viral DNA of PvB19 and its correlation with hemoglobin levels and human erythropoietin dosage in patients with kidney failure under hemodialysis treatment.
Patients and Methods: All patients had chronic renal failure and were undergoing hemodialysis. The detection of the viral DNA was performed by qualitative polymerase chain reaction (PCR). The PCR reaction was carried out using one specifically designed primer set for PvB19 that encoded for a structural capsid protein (VP1). Erythropoietin resistance (ER) was diagnosed using the European Best practice Guidelines as stated by Hay N (Nephrology, 2002).
Results: A total of 58 samples of peripheral blood were obtained from patients with chronic kidney failure. Mean age was 59.41 (23 to 85 years) and mean Hemoglobin levels were 10.1 g/dL (7–14). Iron was normal in all patients and no patient had active infections. Fifty two patients (89.65%) were receiving recombinant erythropoietin. Among all the samples, 3 cases met ER criteria but only in 1 of them did we find the viral DNA (1.72%). Three cases (5.17%) positive to the viral DNA were found in the non erythropoietin resistance group.
Conclusion: Although ours is a small group and we are still gathering samples, up until now, no relation appears to exist between the presence of PvB19 genetic material and ER.
Pulmonary Hypertension in Adults and Children with Sickle Cell Disease in Nigeria: Prevalence, Clinical Characteristics and Role of Endemic Tropical Infections.
Authors: Aliyu Zakari Y. , Vandana Sachdev , Mamman Aisha I. , Babadoko Aliyu , Akpanpe Peter , Attah Ester , Suleiman Yusuf , Aliyu Nurudeen , Yusuf Jamilu , Mendelsohn Laurel , Coles Wynona , Danbauchi Solomon , Taylor James , Castro Oswaldo , Gordeuk Victor R. , Blackwelder William C. , Kato Gregory J. , Gladwin Mark T. 
Published: April 2020

Abstract
Pulmonary hypertension has a prevalence of 30% in patients with sickle cell disease (SCD) in the United States with mortality rates of 40% at 40 months after diagnosis. The global burden of SCD is highest in sub-Saharan Africa where more than 200,000 children are born with the disease annually. The prevalence of pulmonary hypertension among individuals with SCD in Africa has not been previously reported. We performed Doppler echocardiographic assessments of pulmonary-artery systolic pressure in 206 consecutive hydroxyurea sickle cell patients at steady state in Nigeria, West Africa (101 males and 105 females; age range 10–52, mean [+/−SD] age, 21.5 +/− 7.7 years; 196 homozygous sickle cell and 10 compound heterozygotes SC). A control group consisted of 93 healthy Nigerians. Hemoglobin gentotype was determined by electrophoresis and DNA sequencing. Pulmonary hypertension was defined prospectively as a tricuspid regurgitant jet velocity (TRV) of at least 2.5 m per second. We collected clinical data on the patients and controls, and blood specimens for clinical laboratory measurements. Doppler-defined pulmonary hypertension occurred in 25% of sickle cell patients (21% with TRV 2.5 – 2.9 m/sec, 4% with TRV ≥ 3 m/sec). The presence of pulmonary hypertension was inversly associated with age (p=0.04) and hemoglobin (p=0.0016), and positively associated with reticulocyte count, serum levels of lactose dehydrogenase (p=0.03), creatine kinase, and blood urea nitrogen and systolic (p=0.03) and diastolic blood pressure (p=0.002) in bivariate analyses. In a multivariate linear regression model age, diastolic blood pressure and blood urea nitrogen had significant independent associations with pulmonary hypertension. There were no significant associations of HIV/AIDS, hepatitis B and C co infections and malarial parasitemia rate with pulmonary hypertension. Our findings suggest that pulmonary hypertension is common among sickle cell patients in Africa and it appears to be a complication of chronic hemolysis and vasculopathy. The prevalence of pulmonary hypertension decreases with age in Nigerian SCD patients, in sharp contrast to U.S. SCD patients, who demonstrate increasing prevalence with age. The public health implications of this finding are significant considering the potential number of individuals at risk for this complication. Large prospective cohort studies to determine the outcome of pulmonary hypertension in sickle cell patients in Africa are needed.