bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of a New Assay Based on a FV-Dependent Prothrombinase Activator for the Detection of Activated Protein C Resistance Phenotype.
Authors: Wu Jogin , Quehenberger Peter , Foltyn Katherine , Dillard Patricia 
Published: October 2019

Abstract
Activated protein C (APC) resistance is the most frequent hereditary defect associated with deep venous thrombosis. Major cause of APC resistance phenotype is due to a point mutation of factor V (Factor V Leiden). A new clotting assay, Pefakit® APC-R Factor V Leiden (Pentapharm Ltd., Switzerland) for the detection of APC resistance phenotype was evaluated at two tertiary care hospitals, Duke University Medical Center, USA (Duke) and University of Vienna, Austria (Vienna). Samples of 242 subjects from Duke and 187 subjects from Vienna were included in the study among patients who were subjects for thormbophilia screening. The Pefakit® method is based on clotting time measurement triggered by a prothrombin activator added to a mixture of patient plasma diluted with factor V deficient plasma with and without APC. Robustness and specificity of the assay is enhanced by elimination of possible disturbing influence by factors upstream the coagulation cascade and heparin interference is precluded up to heparin level of 2 IU/ml by heparin inhibitor added to the regent. A similar, FDA approved commercial APC-R kit (COATEST of IL) was used for method comparison. Patients with elevated factor VIII (n=11), Coumadin (n=23), Lupus Anticoagulant (n-14), protein C deficient (n=7), protein S deficient (n=9), AT III (n=6) and women with pregnancy (n=9) were included in Duke study and no interference were found in phenotype. Using PCR/FRET DNA method as reference method the Pefakit® method provided 100 % sensitivity and 100 % specificity for the Vienna study and 99.0 % sensitivity and 98.6 % specificity for the Duke study and the COATEST provided 97.1 % specificity and 93.2 % sensitivity with the Duke study. Using two levels of genotype controls both studies showed similar intra and inter-assay precision (less than 6 % for the Vienna study and 9 % for the Duke study) as compared with the gold standard IL APC-R COATEST kit (less than 5 % CV). Of great interest one false positive sample from the Duke study is under investigation due to that the functional detection of the assay is supposed to detect other FV mutations leading to APC-R phenotype as well. Reasons that cause the other two false negative results for the Duke study are still unknown and under investigation. Both studies showed that the Pefakit® is simple and rebust assay. Both wild type and heterozygous groups have much higher ratio as compared with the reference method in differentiating them from homozygous phenotype.
Figure Figure
Incidence of Pulmonary Thromboembolism (PTE) and New Guidelines for PTE Prophylaxis in Japan.
Authors: Kobayashi Takao , Nakamura Mashio , Sakuma Masahito , Yamada Norikazu , Sakon Masato , Fujita Satoru , Seo Norimasa 
Published: October 2019

Abstract
Venous thromboembolism (VTE) has been on the rise in Japan in recent years, together with the Westernization of Japanese eating habits. According to the investigation by Editorial Committee on Guideline for Prevention of Venous Thromboembolism by Japanese Society of Anesthesiologists in 2002, 369 cases of pulmonary thromboembolism (PTE) were registered among 467 General Hospitals and University Hospitals. The rate of perioperative PTE is estimated to be 0.044% (369/837540), and the fatal rate among clinical PTE was 17.9%. 36% of the cases occurred in Orthopedics, 22% in general surgery and 10% in Obstetrics and Gynecology. 59% of the cases did not received prophylaxis, and 52% of the cases were restricted mobility. The incidence of clinical PTE after general surgery was 0.16% from 1994 to 2000 in Japan. Fatal rate among clinical PTE was 21.3%, and fatal rate among all patients was 0.04%. The incidence of clinical PTE after total hip joint replacement surgery among 46 papers between 1990 and 2001 in Japan was 1.1%, and that of fatal PTE was 0.15% among all patients. Patients of PTE were investigated between 1991 and 2000 among 68 University Hospitals and 34 General Hospitals in Japan. 76 cases of PTE occurred in Obstetrics, and 13.2% of them were fatal. The incidence of PTE was 0.003% in vaginal deliveries and 0.06% in cesarean sections. 178 cases of PTE occurred in Gynecology, and 13.5% of them were fatal. The incidence of PTE was 0.03% in benign diseases and 0.42% in malignant diseases. However, the incidence of PTE in Japan is considered to be at least one level lower compared with Western populations according to ACCP guidelines. Furthermore, low molecular weight heparin (LMWH) is not covered by health insurance and is contraindicated for pregnant women still now in Japan. Then, we established Japanese guidelines for VTE prophylaxis according to Japanese clinical evidences of PTE in 2004. We classified four risk groups according to ACCP guidelines. Recommended prophylaxis is early ambulation for low risk group, graduated compression stocking (GCS) or intermittent pneumatic compression (IPC) for moderate risk group, IPC or low dose unfractionated heparin (LDUH) for high risk group, and LDUH + IPC or LDUH + GCS for highest risk group. And, risk group should be raised one rank in cases with any additional risks, such as obesity, advanced age, pregnancy, operation time, and other complications. Recommended prophylaxis in this guideline may be minimum one, because the purpose of this guideline is to penetrate the awareness and education of VTE throughout Japan. Fortunately, the management fee for PTE prophylaxis was established and covered by health insurance in April 2004. However, after accumulation of further evidences and application of pharmacological agents, such as LMWH, we will establish the advanced guidelines in the future.
A Role for SCL in Developing Bone? the SCL +19 Core Enhancer Targets Expression to Cells of Developing Bone, Osteocytes, Chondrocytes and Bone Lining Cells.
Authors: Silberstein Lev , Pimanda John E. , Piltz Sandie , Delaney Liz , Oldham Scott , Kallianpur Asha , Dominici Massimo , Brandt Stephen , Gottgens Berthold , Green Anthony 
Published: October 2019

Abstract
The transcription factor SCL/TAL1 is essential for haematopoiesis and vascular development in the embryo. It is also expressed in the developing skeletal and central nervous systems, but lethality of SCL null mice has precluded detailed analysis of the role of this gene in these tissues.We have previously demonstrated that the SCL +18/19 enhancer directs reporter gene expression to haemangioblasts, endothelium and haematopoietic progenitors. Using human placental alkaline phosphatase as a reporter gene, we now report that the +19 core enhancer also directs expression to osteocytes, articular chondrocytes and bone lining cells in adult mice. In E14.5 embryos, the transgene is expressed in cells within the cartilaginous template of future bone and in the perichondrium. The pattern of expression of the transgene resembles that of endogenous SCL RNA and protein expression in age matched embryos. At E16.5, transgene-positive cells are seen within ossification centres. Activity of the enhancer during osteogenesis was observed both in bones formed through intra-membranous (e.g. cranial vault) and endochondral ossification (limbs and axial skeleton). We found that the murine pre-osteoblast cell line, MC3T3, expresses endogenous SCL. Stable transfection of these cells with luciferase reporter constructs that include the +19 core enhancer produce a 5-fold increase in luciferase activity. There is a progressive reduction in SCL RNA expression in these cells during osteogenic differentiation. This is consistent with a previous report that expression of SCL protein is down-regulated in skeletal tissues of more advanced stage embryos, and with our finding that the MLO-Y4 cell line, which phenotypically and functionally resembles mature osteocytes, does not express SCL. We addressed the issue of SCL function in bone formation using in-vitro differentiation of embryonic stem (ES) cells. Wild type and SCL−/− J1 ES cells did not differ in their ability to form bone nodules. These data demonstrate that SCL is not required for bone specification, but do not exclude a role for SCL in regulating bone formation in vivo. The SCL +19 core enhancer directs expression to haematopoietic progenitors and endothelium together with their putative precursors, haematopoietic endothelium and haemangioblasts. Our demonstration that the +19 core enhancer also targets expression to osteogenic cells is consistent with recent data suggesting that haematopoietic and osteogenic cells share a common precursor (Olmsted-Davis, EA et al PNAS 2003, Dominici M et al PNAS 2004) and that osteoblastic bone lining cells are intimately associated with haematopoietic stem cells (Calvi LM et al Nature 2003; Zhang J et al Nature 2003).
Erythropoietin Did Not Activate the JAK/STAT or the ERK1/2 Pathway in the Human Colorectal Cell Line, HT-29.
Authors: Tullai Jennifer , Chitikila Carmelata , Gechtman Ze’ev , Wang Xiwei , Jolliffe Linda , Farrell Francis 
Published: October 2019

Abstract
Erythropoietin (EPO) is the cytokine essential for erythropoiesis; however, the expression of EPO and EPO receptor (EPOR) is not limited to cells of erythroid lineage. Erythropoietin and EPOR expression has been localized to numerous non-hematopoietic cells and tissues including endothelial, neuronal and ovarian. Moreover, several tumor types have been shown to express both EPO and EPOR and display increased expression upon hypoxia. To shed light on the potential biological role of EPOR and tumor cells we characterized the response of human colon carcinoma HT-29 cells to EPO. Functionality of the erythropoietin receptor was assessed by radio-labeled ligand binding, cellular proliferation/signaling, and gene expression using DNA microarrays. Receptor binding experiments using [125I]-EPO did not reveal measurable EPO binding activity present on the surface of HT-29 cells under both normoxic and hypoxic conditions. Moreover, EPO failed to induce cellular proliferation or an increase in the phosphorylation state of STAT5, EPOR or ERK1/2 under normoxic or hypoxic conditions at supra-pharmacological levels (25 IU/ml). Gene expression analysis revealed no significant change in gene expression in response to EPO (5 IU/ml) under normoxic conditions. On the other hand, over 347 genes exhibited greater than a 1.5 fold change in gene expression when cells were cultured under hypoxic conditions (1 % O2). When EPO was administered to cells in the hypoxic state, 36 additional genes were observed (9 and 27 up-regulated or down-regulated, respectively). That HT-29 cells exhibit minor transcriptional changes in response to EPO raises the possibility that EPO may signal in HT-29 cells. However, the mechanism for this response is not through the previously described EPO/EPOR signal transduction pathway. This conclusion is supported by the apparent lack of EPO receptor expression on the cell surface. These results suggest that tumor microenvironment, e.g., hypoxia, exerts a greater effect than that seen by exposure to erythropoietin.
Early Results Suggest That Epoetin Alfa 60,000 U Every 2 Weeks Improves Hemoglobin in Patients with Cancer Not Receiving Chemotherapy or Radiotherapy.
Authors: Shasha Daniel , Henry David H. , Williams Denise 
Published: October 2019

Abstract
Epoetin alfa has been shown to increase hemoglobin (Hb), decrease transfusion requirements, and improve quality of life (QOL) in patients receiving chemotherapy (CT) and/or radiotherapy (RT). However, the efficacy of epoetin alfa in cancer patients receiving neither CT nor RT is not as well characterized, particularly at doses less frequent than 3 times weekly. The objective of this open-label, nonrandomized, multicenter pilot study was to evaluate epoetin alfa 60,000 U subcutaneously (SC) every 2 weeks (Q2W) in 50 anemic patients with cancer who were not receiving CT or RT. The primary efficacy endpoint is the proportion of patients achieving a hematopoietic response (HR; Hb increase ≥2 g/dL from baseline and/or Hb increase to ≥12 g/dL at any scheduled visit) independent of transfusion within 28 days. Patients with histologically confirmed nonmyeloid malignancy, Hb ≤11 g/dL, and who had not received CT in the previous 8 weeks or RT in the previous 4 weeks were enrolled. Patients were permitted to receive hormonal therapy, androgen deprivation therapy, and/or immunotherapy. Patients were to receive epoetin alfa 60,000 U SC Q2W, with escalation to 80,000 U Q2W after 4 weeks if Hb increased ≤1 g/dL. Dose will be titrated to maintain Hb ≤13 g/dL; all patients were to receive oral ferrous sulfate 325 mg daily. Patients will be treated for up to 12 weeks, with a 4-week follow-up after last dose of study drug. As of July 2004, 18 patients are evaluable for efficacy (modified intent-to-treat, ie, all enrolled patients who received ≥1 dose of study drug and had ≥1 postbaseline Hb or transfusion evaluation) and 18 for safety (all enrolled patients who received ≥1 dose of study drug). Mean age was 74.4 ± 8.7 yrs, mean baseline Hb was 10.0 ± 1.0 g/dL (n=18), and 7/18 were men. Of these patients, 7 completed 4 weeks, 3 completed 8 weeks, and 1 completed 12 weeks on study. The HR of these patients was assessed regardless of how many weeks of study they completed at the time of this interim analysis; HR was 44.4%. Hb increased 1.4 ± 0.7 g/dL (n=11) after 4 weeks and 1.2 ± 1.4 g/dL (n=4) after 8 weeks of treatment. Adverse events were limited to 1 patient, who experienced back pain and epistaxis. No patients died during the study, and no thrombotic vascular events were reported. Early results of this study suggest that epoetin alfa 60,000-80,000 U SC given every other week is well tolerated and appears to effectively increase Hb in anemic cancer patients not receiving CT or RT. The study is currently ongoing.
Usefulness of CD10 Study by Multi-Colour Flow Cytometry in Angioimmunoblastic T-Cell Lymphoma.
Authors: Baseggio Lucile , Berger Francoise , Carret Josiane , Thieblemont Catherine , Morel Dominique , Magaud Jean-Pierre , Felman Pascale 
Published: October 2019

Abstract
Angioimmunoblastic T-cell lymphoma (AITL) is a distinct clinicopathological entity among peripheral T-cell lymphoma in the WHO classification. Whereas antigen “loss” or “deletion” of one or several pan-T cell antigens is a hepful feature of neoplastic lymphocytes in many T-cell lymphomas, no specific immunophenotypic patterns were available to recognize the tumour T-cells of AITL until recently. Indeed, Attygalle et al. reported that in this disorder neoplastic T-cells can be recognized by the aberrant expression of CD10 using immunochemistry in lymph nodes as well as in the involved extranodal sites. Lee et al. has also confirmed this specific phenotypic feature in cell suspension of lymph nodes using flow cytometry (FCM) in 3 cases of AITL. Here, we evaluated the CD10 expression by T cells in patients with AITL using four-colour FCM. The present study included lymph nodes (LN, n=10), peripheral blood (PB, n=5), bone marrow (n=1) and skin (n=1) samples from 13 patients with a diagnosis of AITL and with available cytologic histologic, immunologic and molecular data. Lymph nodes of reactive hyperplasia (n=13), B-cell lymphoma (n=23), other T-cell lymphoma (n=6) and peripheral blood from healthy donors (n=18) were used as control group. According with previous immunohistochemistry results, a fraction of T-cells expressed CD10 (using a level of at least 5% of all CD5+ cells) in 9/10 AITL lymph nodes with a mean number of 18%. Interestingly, among these 9 cases, 5 could be studied in peripheral blood also and all cases showed a fraction of T-cells expressing CD10, whatever be the lymphocytosis (median 1.1 109/l range 0.82 to 11.32 109/l). In three of these cases, tumoral T-cells presented also lack of surface CD3. In two cases of AITL diagnosed in LN, the aberrant CD10 expression by T-cells was found in bone marrow and skin, respectively. In the control group, T-cells were CD10 negative using the cut-off of 5%.
In conclusion, we demonstrate that the assessment of CD10 expression by neoplastic T-cells can be achieved by multi-colour FCM in lymph nodes and involved extranodal sites. Our results are concordant with the statement of Attygalle that CD10 expression by T-cells can be used as a marker of both malignancy and AITL type. In addition, this is to our knowledge the first description of circulating CD10 neoplastic T-cells in AITL. Further study with a larger series of patients is required to confirm these data, to standardize the cut-off of positivity and to evaluate the sensibility of FCM versus immunohistochemistry.
Figure Figure
Intensified Chop (CHOP14) Plus Rituximab in the Treatment of Elderly Patients with Aggressive and High Risk Non Hodgkin’s Lymphoma (NHL).
Authors: Rigacci Luigi , Nassi Luca , Alterini Renato , Carrai Valentina , Bernardi Franco , Martini Varesco , Bernardeschi Paolo , Longo Giovanni , Bosi Alberto 
Published: October 2019

Abstract
The results of treatment of aggressive NHL in elderly patients is relatively poor. The use of protocol specifically devised for eldery patients can reduce hematological and extrahematological toxicity but yield worst results, particularly in high-risk IPI patients. Recentely Coiffier et al. have showed that the standard therapy for patients over 60 years with aggressive lymphoma was rituximab plus CHOP. With the aim to improve dose intensity in elderly large cell lymphoma patients and intermediate-high or high-risk IPI score we planned treatment with rituximab 375 mg/sqm on day 0 and CHOP therapy on day +2 given every 2 weeks, with G-CSF starting from day + 7 to + 11 and a profilaxis with cotrimoxazole twice weekly and itraconazole 100 mg day. With an intention to treat analysis we have included in the study, from June 2002 to June 2004, 25 patients with the following characteristics: IPI score 2 in 8 patients (32%), score 3 in 10 (40%) and score 4–5 in 7 (28%). Two patients had diagnosis of follicular grade III lymphoma and the other 23 diffuse large B-cell lymphoma. Eighteen patients (72%) had abnormal value of LDH and 18 patients had one or more extranodal sites involved. The median age was 64 years (60–76), seven patients (28%) were stage II, four (16%) were stage III and fourteen (56%) were stage IV. The aims of the present study were: 1) feasibility 2) toxicity 3) response to therapy. Twenty-two patients completed the estabilished 6 cycles and were evaluable, one patient stopped the treatment after three cycles due to infections the remaining two patients are in treatment. Six patients delayed one cycle and one patient two cycles, due to trombocytopenia or neutropenia. Only eight out 135 cycles were delayed (6%). Sixteen out 23 (70%) patients obtained complete remission (CR) and after a median follow-up of 11 months (1 – 22 months) the disease-free survival was 79%, three patients (19%) relapsed. Two of these patients died rapidly after relapse and one obtained a second CR. Three out 6 patients with a partial response obtained a CR after a salvage therapy, two died and one is still in therapy. With a median follow-up of 15 months (3–25 months) the overall survival was 78%. No remarkable extrahematological toxicity was observed, in particular, as regards cardiotoxicity, we did not observe significant variation of left ventricular ejection rate in comparison with the diagnostic ones, except one patient who, after the sixth cycle, presented a significant reduction of left ventricular ejection which rapidly recovered after he was admitted at hospital. In conclusion we observed that dose intensified CHOP with rituximab was feasible and lack of severe toxicity in elderly patients with aggressive and high risk prognosis lymphoma.
Modified Magrath IVAC Regimen as Second-Line Therapy for Relapsed and Refractory Aggressive Non-Hodgkin Lymphoma in Developing Countries: The Experience of a Single Center in Brazil.
Authors: Pracchia Luis F. , Pereira Juliana , Belesso Marcelo , Beitler Beatriz , Chamone Dalton A. 
Published: October 2019

Abstract
In this retrospective study we described the response and toxicity of a modified Magrath IVAC (mIVAC) regimen in 25 patients with refractory/relapsed aggressive non-Hodgkin lymphoma (NHL). The mIVAC consisted of ifosfamide 1,500mg/m2 (one-hour infusion beginning at 9:00; D1 to D5), mesna 300mg/m2 (bolus at hours 9:00, 13:00, 17:00; D1 to D5), citarabine 2,000 mg/m2 (two one-hour infusions beginning at 8:00 and 16:00; D1 and D2) and etoposide 60 mg/m2 (one-hour infusion beginning at 10:00; D1 to D5). Treatment was repeated every four weeks for a maximum of six cycles. Patients who achieved partial remission or complete remission after at least three courses were offered autologous stem cell transplantation (ASCT), if eligible. The median age was 37 years (range 18 to 59 years). Twenty-two (88%) patients had diffuse large B-cell lymphoma, fourteen (56%) had relapsed disease and 10 (40%) were considered high-intermediate and high risk by age-adjusted International Prognostic Index. The overall response rate was 68% (95% CI: 46%–90%). A total of 64 cycles were given, with a median of three courses per patient. Grade 3/4 neutropenia was observed after 85,6% of the courses, and grade 3/4 thrombocytopenia was observed after 87,5% of the courses. Grade 3/4 neutropenic fever occurred after 28% of the courses. Non-hematologic toxic effects were rare, predominantly grade 1/2. No toxic deaths were observed. Fifteen (88%) of the 17 responding patients underwent ASCT. With a median follow-up of 14 months, the median overall survival time for mIVAC sensitive patients was 16 months. This regimen may be feasible for patient with relapsed and refractory aggressive NHL in countries with inadequate numbers of hospital beds.