bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pulmonary Hypertension as a Risk Factor for Death in Patients with Sickle Cell Anemia
Authors: Cancado Rodolfo D , Olivato Maria Cristina A , Filho Newton Nunes Lima , Campos Orlando , Chiattone Carlos 
Published: April 2020

Abstract
Pulmonary hypertension develops in most forms of hereditary and chronic hemolytic anemia, including sickle cell disease, thalassemia, hereditary spherocytosis, and paroxysmal nocturnal hemoglobinuria, suggesting that there is a clinical syndrome of hemolysis-associated pulmonary hypertension. Retrospective studies from tertiary care referral centers suggest a prevalence of pulmonary hypertension in adults with sickle cell disease ranging from 20 to 40%. Despite the fact the elevations in pulmonary artery pressures are slight, morbidity and mortality are high. In adult sickle cell anemia patients, pulmonary hypertension is emerging as a major risk factor for death. We performed Doppler echocardiographic assessments of pulmonary-artery systolic pressure in 80 consecutive patients (20 men and 60 women; mean [±SD] age, 30 ± 10.8 years) between 1/20/2006 and 1/20/2008. The genotype on the basis of hematologic and hemoglobin characteristics was hemoglobin SS in all patients. Pulmonary hypertension was prospectively defined as a tricuspid regurgitant Jet velocity (TFJV) of at least 2.5 m per second. Patients were followed for a mean of 18 months (6–24 months), and data were censored at the time of death or loss to follow-up. Doppler-defined pulmonary hypertension occurred in 37.5 percent of patients (30/80). Multiple logistic-regression analysis, with the use of the dichotomous variable of a tricuspid regurgitant jet velocity of less than 2.5 m per second or 2.5 m per second or more, identified age, female sex, deferasirox therapy, left ventricular mass index, pulmonary artery systolic pressure, reticulocytes, white-cell count, platelet count, lactate dehydrogenase (a marker of hemolysis), blood urea nitrogen, creatinine, uric acid and self-reported history of cardiovascular complication, billiary stones, retinopathy and acute chest syndrome, as significant independent correlates of pulmonary hypertension. The hemoglobin level, fetal hemoglobin level, hydroxyurea therapy and serum ferritin level were unrelated to pulmonary hypertension. Hazard rate for death according to the TFJV of at least 2.5 m per second, as compared with a velocity of less than 2.5 m per second, was associated with an increased risk of death (0.00 versus 2.54; P=0.998). Mortality rate in 24 months was 6.7% (2/30) for patients with TRJ velocity ≥ 2.5 m/sec versus 0.0% (0/50) for patients without pulmonary hypertension. Pulmonary hypertension, diagnosed by Doppler echocardiography, is common in adults with sickle cell disease. It appears to be a complication of chronic hemolysis, is resistant to hydroxyurea therapy, and confers a high risk of death. Large trials evaluating the effects of treatment for pulmonary hypertension in the sickle cell anemia population are indicated.
ERG, EVI1 and PRAME Are Relevant Markers of Treatment Response and Survival, and They Could Be Useful for Improving the Risk-Stratification in Citogenetically Normal Acute Myeloid Leukemia (CN-AML)
Authors: Santamaría Carlos , Chillón Carmen , García-Sanz Ramon , Pérez Cristina , Ramos Fernando , de Coca Alfonso García , Alonso Jose- Maria , Giraldo Pilar , Bernal Teresa , Queizán José-Antonio , Rodríguez José Nicolas , Fernández-Abellán Pascual , Barez Abelardo , Peñarrubia María-José , Amigo Mari Luz , García-Sancho Alejandro Martín , Pozas Miguel Angel , Balanzategui Ana , Díaz-Mediavilla Joaquin , San Miguel Jesús F. , González Marcos 
Published: April 2020

Abstract
Several mutations and aberrant gene expression have been proposed as prognostic factors in cytogenetically normal acute myeloid leukemia (CN-AML), but few studies have analyzed the clinical value of several genes in a large group of CN-AML patients. In 121 de novo CN-AML included into the Spanish PETHEMA therapeutic protocols, we evaluated FLT3 and NPM1 mutations and BAALC, ERG, EVI1, MLL-PTD, MN1, PRAME, RHAMM and WT1 RNA levels. Ninety-one patients achieved CR with standard induction therapy while 20/111 (16.5%) were refractory to treatment. This latter subgroup showed higher ERG (media 1.0±0.8 vs. 0.6±0.6;p=0.012) and lower PRAME (media 28.8±52.9 vs. 1640.6±6102.0;p=0.014) levels than no-refractory patients. Moreover, high ERG expression (defined by median value) was associated with shorter OS at 2-years (38% vs. 61%;p=0.012) and RFS at 2-years (38% vs. 60%;p=0.005). By contrast, high PRAME levels (defined by 75th percentile) was related to longer OS (61% vs. 48%;p=0.029) and RFS (74% vs. 43%;p=0.016). Additionally, high EVI expression (defined by 75th percentile) was associated with a poorer OS (37% vs. 56%;p=0.049). The same results in OS were observed in patients with high BAALC RNA levels (median value; 41% vs. 62%; p=0.049). Interestingly, ERG, EVI1 and PRAME markers improved the current prognostic classification of CN-AML based on FLT3 and NPM1 status. Therefore, intermediate risk patients (FLT3-ITD/NPM1mut and FLT3wt/NMP1wt) with low ERG, low EVI1 or high PRAME achieved long OS and RFS, similar to the favorable FLT3wt/NPM1mut subgroup. By contrast, the opposite expression subgroup (high ERG, high EVI1 or low PRAME) showed OS and RFS similar to the adverse FLT3-ITD/NPM1wt subset. These results allowed identify clearly two risk-subgroups for OS and RFS, based on three molecular markers (FLT3, NPM and one of the 3 proposed genes). In conclusion, we demonstrated that ERG, EVI1 and PRAME are relevant prognostics markers in CN-AML and they could improve their risk-stratification.
Analysis of γH2AX and NHEJ Signaling as Molecular Determinants for GOSensitivity in AML
Authors: Hååg Petra , Lindberg Marita Lagergren , Kanter Lena , Lewensohn Rolf , Stenke Leif , Viktorsson Kristina 
Published: April 2020

Abstract
Most patients with acute myeloid leukemia (AML) display an initial response to high dose chemotherapy but a majority develops resistance and relapse within 1–2 years. The targeted drug gemtuzumab ozogamicin (GO) has shown promising clinical effects. Consisting of a monoclonal CD33 antibody coupled to a toxin, calicheamicin, GO enters the cell after binding to the CD33 antigen, preferentially expressed on myeloid cells. Calicheamicin is then released from the antibody and causes multiple signaling effects resulting in cell death. We have previously shown that GO-induced apoptotic signaling in AML cells involves activation of the Bcl-2 proteins Bax and/or Bak. In this study we aimed to clarify if altered DNA repair or DNA damage signaling are molecular determinants of GO-sensitivity in AML. Moreover, attempts were made to connect the upstream DNA double stranded break (dsb) signaling to GO-induced activation of Bax and Bak. Therefore, we analyzed GO-induced DNA damage signaling in GO responsive (HL60) and resistant (KG1a) AML cells. We can show that GO-resistant cells display a higher basal activity of γH2AX and DNA-PKcs and hence a higher DNA repair capacity. Moreover, by treating AML cells with clinically relevant doses of GO we have observed an increased level of both γH2AX and phosphorylated DNA-PKcs which indicates the prevalence of DNA dsbs. This activation was observed in HL60 as well as in KG1a cells, in vitro. Thus, our data suggest that resistance to GO is not due to lack of DNA dsbs induction, but is rather influenced by the DNA dsbs signaling to the downstream apoptotic components. Key activators downstream of DNA-PK but upstream of Bak/Bax, including c-Abl, caspase-2, Bid, Bad, Bim, Bcl-2 and Bcl-xL, are therefore currently being analyzed. In conclusion, we show that GO induced DNA dsbs in both GO sensitive and GO resistant AML cells in vitro, and that resistance to GO treatment is at least in part due to factors downstream of DNA-PK but upstream of Bak/Bax activation.
Prevalence and Clinical Characterization of CD20 and CD22 Expression in Pediatric Precursor B-Cell Acute Lymphoblastic Leukemia
Authors: Rayburg Melissa S , Marmer Daniel , Mo Jun , McMasters Richard , Smolarek Teresa , Lampkin Beatrice , Perentesis John 
Published: April 2020

Abstract
Immunophenotypic classification of leukemia has important therapeutic and prognostic implications. In B-cell malignancies, CD20 and CD22 also represent therapeutic targets. CD20 expression in adult patients with B lineage ALL has been associated with a poor prognosis. The data are conflicting in the pediatric population and may be impacted by the type of therapy employed. A recent study of pediatric patients treated in consecutive St Jude Children’s Research Hospital Total Therapy ALL regimens observed excellent outcomes and no prognostic significance for CD20 expression (Jeha et al. Blood2006, 108:3302–3304). We retrospectively analyzed 50 consecutive patients aged 4 months to 28 years with precursor-B ALL treated with contemporary risk-adapted BFM-based ALL regimens for whom flow cytometric, genetic, and early response data were available. Cases were defined as positive for CD20 and/or CD22 expression if surface expression was identified in more than 20% of leukemic blasts. We found that CD22 was expressed at high levels (68–99%) in all patients evaluated. CD20 expression was positive in 27 (54%) of patients. CD20 expression did not correlate with known NCI prognostic features, including presenting white blood count or age. All 3 patients with BCR-ABL translocation ALL were CD20 positive. Consistent with previously published data, neither of the 2 patients with MLL-AF4 translocation were CD20 positive. There was no association of CD20 expression with trisomy 4/10/17 or TEL-AML1 status. We did not observe an association between CD20 expression and rapid early bone marrow response to therapy at day 8 or 15; 47/50 patients were in remission at day 29. At a median follow-up time of 48 months 46/50 patients were alive without relapse. These limited data do not suggest a strong association between CD20 expression and known prognostic features or early treatment response in pediatric precursor-B ALL treated with contemporary BFM therapy platforms. However, our findings of frequent expression of CD22 on precursor-B ALL blasts from children supports its consideration as a target for immunotherapy approaches in high risk or relapsed disease.
T-Cell Large Granular Lymphocytic Leukemias/Proliferations and Clinical Implications in a Community Practice
Authors: Mathews Malcolm (Mack) , Gale Peter F. , Silva Marco A.C.P. Da , Bailey Teresa W 
Published: April 2020

Abstract
Clonal T-Cell large granular lymphocytic leukemias/proliferations were diagnosed in eighteen patients in the community of Kingsport, TN, from 2000–2008, based on peripheral blood and bone marrow morphology, flow cytometry, and T-cell receptor gene rearrangement studies. These clonal expansions were associated with anemia, neutropenia, and thrombocytopenia. Some patients required no treatment, while others responded to weekly methotrexate. Associated disease entities included myelodysplastic syndrome, non-hodgkins lymphoma, multiple myeloma, mast cell disease, febrile illness, and rheumatoid arthritis. Acquired autoimmune hemolytic anemia developed in one case. Anemia was a common presenting feature. Splenomegaly was noted in several cases. Clonal rearrangement of T-cell receptor gamma chain genes was confirmatory in all cases. Chronic disease with uncomplicated cytopenias was noted in the majority. Diagnosis should be suspected in patients with neutropenia and anemia with a relative (and at times an absolute) peripheral blood large granular lymphocytosis. Clonality can be assessed with molecular studies, and therapy is reserved for symptomatic disease or transfusion dependency.
A Case Report of Aggressive Adult Neuroblastoma Mimicking Acute Leukemia with Fulminant Course and Fatal Outcome
Authors: Panovska Irina , Ivanovski Martin , Trajkova Sanja , Pivkova Aleksandra , Genadieva-Stavric Sonja , Hadzi-Pecova Liljana , Latifi Arif , Trajkov Dejan , Spiroski Mirko , Cevreska Lidija 
Published: April 2020

Abstract
Neuroblastoma is an embryonic neoplasm of the sympathetic nervous system derived from the primitive neural crest cells. It is the second most common extra cranial malignant tumor of childhood and the most common solid tumor of infancy, comprising 8 to 10 percents of pediatric cancers. Forty percent of patients who present with clinical symptoms at diagnosis are under 1 year of age, and less than 5% with clinical symptoms are over the age of 10 years. The tumor very rarely occurs in adult patient (defined as 20 years of age and older) and usually has an indolent course with poor survival. Up until June 2008, no more than 65 neuroblastomas in adults had been reported in the literature. Majority of the reports describe an indolent and longer course of the disease among adults with different biological characteristics when compared to children, but ultimately poor outcome regardless of the initial stage of the disease. Here, we report a case of aggressive adult neuroblastoma mimicking acute leukemia with fulminant course and fatal outcome. The patient, a previously healthy 38 years old Caucasian male, presented at our department with a 4 months history of hip pain, headaches, malaise, weakness, persistent diarrhea, poor appetite and weight loss. On admission he was febrile, prostrated, pale, fixed in bed. His abdomen was distended with present bowel movements. The liver and spleen were 3 cm and 5 cm below the costal margins respectively. No abdominal mass was palpable. Neurological examination showed right Horner’s syndrome, reduction of the superficial sensitivity on the Th1 and Th2 level and loss of the tendon-muscle reflexes of the lower limbs. He had paraplegia, indicating possible spinal cord compression. The complete blood counts revealed pancytopenia and presence of 11 % peculiar circulating blastoid cells. The LDH was 3552U/l. Under suspicion of acute leukemia bone marrow aspirate and biopsy was performed. Immunophenotyping of the bone marrow mononuclear cells showed presence of around 50% CD45negative, CD56 and CD9 positive cells, indicating a possible involvement of the bone marrow with neuroblastoma. A routine second opinion from a reference pathologist was also interpreted as neuroblastoma and confirmed with positive immunohistochemical staining for NSE, chromogranin, synaptophysin and negative for LCA. A work-up for neuroblastoma was done and investigations confirmed disseminated stage 4 neuroblastoma. In spite of the appropriate treatment with intensive chemotherapy regimen, the patient failed to improve and died of progressive disease. Our case report illustrate the usefulness of including CD56 marker in flow-cytometry acute leukemia assays, especially in cases when CD45 expression is absent, as initial screening tool for recognizing cases of neroblastoma which simulate acute leukemia.
Differential Expression of VEGF Receptors on Human T Cell Subsets
Authors: Lee Stephen A , Corless Nichole L , Lee Dean A. 
Published: April 2020

Abstract
Vascular endothelial growth factors (VEGF) and their receptors (VEGFR) deliver important signals that controlling the angiogenesis of growing, injured, and malignant tissues. Expression of VEGF receptors has been described on a variety of normal and malignant leukocytes, and presumably plays a role in their trafficking to injured tissue. Differential expression of neuropilin-1 (NRP-1) has been described as a marker of regulatory T cells (Treg) in mice, and may be expressed on human thymocytes, but expression on human Tregs has not been confirmed. Moreover, earlier reports of Treg-specific markers in humans were based on the CD4+CD25+ subset, which is not as reliable for Treg identification as it is in mice. We used cell sorting to obtain pure populations of human Tregs (using the more stringent criteria of CD4+CD25+CD125−) and T helper cells (Th) (CD4+CD25−CD127+). We compared expression in these subsets of VEGFR-1, -2, -3, and NRP-1 and -2 using flow cytometry and quantitative RT-PCR, and NRP-1 splice isoforms by RT-PCR using isoform-specific primers. We were not able to identify any NRP-1 expression in Tregs by any of these methods. In contrast, there was marked differential expression of VEGFR-2 (Flk-1) and -3 (Flt-4), both of which exhibited up to 16-fold higher expression in Tregs than in Th. These findings suggest a role for VEGF in recruiting Tregs to sites of tissue growth as a mechanism for blunting potential autoimmunity to neoexpressed antigens, and thus may also explain the accumulation of Tregs in tumors.
Simultaneous Phenotype-Specific Telomere Length Measured by Flow-FISH Using Thermally Stable Quantum Dots
Authors: Kapoor Veena , Hakim Fran , Rehman Najibah , Gress Ronald E , Telford William G 
Published: April 2020

Abstract
The Flow-FISH technique used to measure telomere length by incorporation of fluorescein-labeled oligonucleotide probes complementary to telomeres repeat sequences has been extended to permit simultaneous multiple phenotypic analysis within a single sample. The necessary thermal stability of the fluorescent labeling needed for phenotyping cell populations can be achieved using quantum dots instead of conventional organic fluorophores, which are usually damaged during the high-temperature hybridization treatment and lose their fluorescence, limiting their usefulness for phenotypic analysis. In this study, quantum dots and conventional fluorophores were compared for their ability to survive the 82oC hybridization step necessary for PNA probe annealing. Quantum dots preserved their fluorescence following heat treatment and gave good signal-to-noise ratios when used to detect lymphocyte and monocyte markers (CD3, CD4 and CD14). This was in contrast to phycobiliproteins and low molecular weight fluorochromes such as fluorescein and the Alexa Fluor dyes, which decreased considerably in fluorescence following hybridization. Since addition of fluorescent immunophenotyping complicated an assay already heavily dependent on uniform technique, internal reference standards (previously reported by Lansdorp) were particularly critical in this technique. We used aliquoted frozen calf thymocytes (CT) as an internal standard, determining their telomere length and incorporating them into each sample to monitor tube to tube and day to day assay variability. The telomere length of the CTs and of CD3+CD4+, CD3+CD4− and CD14+, were determined and then corrected by normalization to the known CT telomere length. To validate the overall method, we then measured age dependence on telomere length for monocyte and lymphocyte subpopulations in young, middle-aged and older donors. Results were consistent with results previously reported using traditional Southern blotting and Flow-FISH with no immunphenotyping, suggesting that incorporation of immunolabeling did not adversely affect this technique. This method allows for the rapid, accurate and simultaneous determination of the telomere lengths of different cell populations within a single sample when little amount of sample is available.
Characterization of Circulating Monoclonal B Lymphocytes in Patients with HCV Infection
Authors: Ohtsubo Korenori , Okamura Takashi , Sata Michio , Kawaguchi Takumi , Morishige Satoshi , Takata Yuka , Oku Eijiro , Imamura Rie , Seki Ritsuko , Hashiguchi Michitoshi , Osaki Koichi , Yakushiji Kazuaki , Kanaji Taisuke , Yoshimoto Kohji , Ueno Takato 
Published: April 2020

Abstract
Hepatitis C virus (HCV), being lymphotrophic as well as hepatotrophic, has been reported to induce B-cell proliferative disorders such as mixed cryoglobulinemia and B-cell lymphoma. To investigate the association between HCV and B-cell proliferation, we evaluated the incidence and characteristics of monoclonal B cells in the circulating blood of HCV-infected cases relative to controls with non-HCV hepatic diseases. We first evaluated the surface immunoglobulin κ:λ light-chain ratios of the circulating B (CD19+) cells in 240 HCV-infected cases and 150 controls. Light chain restriction (κ:λ ratio &gt;3:1 or &lt; 1:2) was detected in 7 cases with HCV (2.9%) (Table 1), but was never detected in the controls (p&lt;0.05). As these monoclonal B cells were not identified morphologically, they were analyzed for CD5/CD20 expression. None of them showed the so-called chronic lymphocytic leukemia (CLL)-phenotype cells. The clonal and normal B cells did not differ significantly in their intensity of CD5 expression (Figure 1). The B-cell clonality was confirmed in all 7 cases by polymerase chain reaction (PCR) analysis of the immunoglobulin heavy-chain (IgH) gene rearrangements and the t(14;18) fusion gene was detected in one case. The loss of clonality was observed in 2 cases treated with interferon and in one case treated with splenectomy. The longitudinal study is required to determine whether these circulating monoclonal B cells progress to lymphoproliferative disorders or not.
Table 1. The clinical data of the 7 HCV-infected cases with monoclonal H cells
Case No Age Sex WBC count./ul Lymphocyte count./ul CD19+ cells in lymphocytes. % Light chain κ/λ. CD5+B cell. % Cryoglobulin IFN therapy 1 78 M 2,800 710 26.46 κ 33.59 17.45 (+) (−) 2 65 M 3,400 720 5.41 λ 0.23 15.93 (−) (+) 3 84 M 5,700 3,520 46.68 κ 11.31 6.8 (+) (−) 4 74 F 3,300 1,750 15.01 λ 0.037 46.31 (−) (−) 5 65 F 2,500 580 34.46 κ 8.07 40.17 (+) (−) 6 72 F 2,000 940 25.4 λ 0.198 38.77 (−) + 7 66 M 4,200 1,380 9.73 λ 0.41 3.53 (+) (−)
Figure 1. The three groups did not differ significantly in the frequency of CD5+ B cells (mean±SEM: group1, 24.14±6.55; group2, 30.27±1.69; group3, 28.53±1.93). Figure 1. The three groups did not differ significantly in the frequency of CD5+ B cells (mean±SEM: group1, 24.14±6.55; group2, 30.27±1.69; group3, 28.53±1.93).