bluuebunny/crossref_metadata_2025_split huggingface dataset:
RITA Activates p53 Function in Chronic Lymphocytic Leukaemia Cells and Induces Apoptosis.
Authors: Nahi Hareth , Merup Mats , Möllgård Lars , Selivanova Galina , Paul Christer 
Published: April 2020

Abstract
The tumor suppressor gene p53 is the most commonly mutated gene in solid tumors. Although less common in hematologic malignancies, about 10–20% of CLL cases carry a p53 mutation. Recently, the compound RITA (Reactivation of p53 and Induction of tumor cells Apoptosis) has been shown to able to bound to p53 and induce its accumulation in tumor cells, its seems to have a cytotoxic effects in tumors with wild type p53.
Peripheral blood or bone marrow was collected from 17 patients with B-CLL, 6 samples wit deleted p53 and 11 without. The cryopreserved samples were incubated in 2 and 3 days with RITA and fludarabine alone or RITA and fludarabine in combination. The cells (3.0x105 cells/ml) were incubated in a medium consisting of RPMI 1640, glutamax, 10% PBS (Phosphate Buffered Saline). Cell viability was assessed after incubation by a bioluminescence method measuring cellular ATP. RITA induced dose and time dependent cell toxic effects in all B-CLL cells but was most effective in those with wild type p53. The combination of 10μM RITA and 1μM of fludarabine resulted in a very high significant synergistic effect. RITA induced dose dependent cell toxic effects in both B-CLL cells with p53 mutation and those without. In the non mutated cells, the main viability compared to unexposed controls after two days of incubation was 54 % (range 20–88 %) at 10μM, 42 % (13–83 %) at 20μM and 31 % (6–870 %) at 50μM. In the patient with a confirmed p53 mutation, the cell viability values were 70 %, 56 % and 30 %, respectively. The combination of 10μM RITA and 1μM of fludarabine decreased the cell viability with an average of 56 percentage points compared to fludarabine alone. We conclude that RITA induces cytotoxic effect in B-CLL and that CLL cells without p53 deletion are more sensitive in vitro. (Preliminary results suggest that RITA cause accumulation of p53 and cause massive apoptosis) Determination of p53 status and apoptosis in the tested patients before and after exposure to RITA is currently performed and the results will be presented at the meeting.
A New Human CD20 Antibody for Improved Killing of CD20-Transgenic T Cells for Allogeneic Stem Cell Transplantation.
Authors: van Meerten Tom , Mackus Wendy J.M. , Parren Paul W.H.I. , van de Winkel Jan G.J. , Hagenbeek Anton , Ebeling Saskia B. 
Published: April 2020

Abstract
Allogeneic stem cell transplantation together with donor lymphocyte infusion represents the best curative option for many haematological malignancies. Donor T lymphocytes play a central role in the elimination of leukemic cells (Graft versus Leukemia, GvL) and acceptance of the graft. Unfortunately, these donor T cells also respond to the patients’ own tissue, resulting in Graft versus Host Disease (GvHD).
In a promising strategy to control GvHD, while at the same time maximally utilizing the GvL effect, genetically-modified donor T cells, expressing a suicide gene, are selectively eliminated in vivo in case of severe GvHD. Genetic transfer of the human CD20 molecule to donor T cells prior to transplantation permits selective elimination via CD20-reactive reagents, such as the widely used chimeric Ab rituximab (RTX). RTX activates complement (CDC), and recruits effector cells (ADCC) for elimination of CD20-positive cells.
Using a novel experimental system we previously showed that the efficiency of RTX-mediated killing of CD20-positive T cells clearly depends on the level of CD20 expression. As a consequence, RTX is not able to kill all genetically modified CD20+ T cells.
Here we investigated whether a novel fully human CD20 Ab, HuMab 7D8, would be better in activating complement and/or effector cells. Our results show HuMab 7D8 to be more powerful in activating complement in CDC-assays. To obtain a similar percentage cell kill compared with RTX, a 100-fold lower dose of HuMab 7D8 was needed and a 4 times lower human serum concentration. To study the effect of the number of CD20 molecules per cell we used CEM-CD20 clones (N=26), all with different CD20-expression levels, in an assay in which all other parameters were kept constant. The results show that HuMab 7D8 is significantly stronger in inducing CD20-mediated cell kill compared to RTX. Especially cells expressing very low levels of CD20, which are completely resistant to RTX-induced CDC, are lysed with high efficiency by HuMab 7D8 and complement. In contrast to RTX, HuMab 7D8 was able to eliminate 100% of transduced T cells in the presence of human serum, and effector cells. At present, studies with CD20-luciferase positive human T cells are conducted in mice to study the in vivo effect of HuMab 7D8 and RTX. These results demonstrate that the CD20 molecule represents an attractive suicide molecule and CD20-transgenic T cells can with high efficiency be eliminated by HuMab 7D8.
Genetically Modified Her2-Specific T Cells Recognize Low and High Her2 Expressing Breast Cancer Cells.
Authors: Ahmed Nabil M. , Ratnayake Maheshika , Pule Martin , Rooney Cliona M. , Heslop Helen E. , Gottschalk Stehpen M. 
Published: April 2020

Abstract
Background: Only patients with breast cancers expressing high levels of Her2 benefit from anti-Her2 monoclonal antibodies such as trastuzumab. In the present study we investigated in vitro, if chimeric T-cell receptors (TCR), which combine the antigen-specificity of monoclonal antibodies with the effector function of T cells can overcome this limitation.
Material and Methods: T cells from healthy donors were transduced with retroviral vectors containing the Her2-specific chimeric TCR gene with a CD28-zeta signaling domain (Her2-CD28-zeta). The specificity of the genetically modified T cells was determined by their ability 1) to kill breast cancer cell lines in cytoxicity assays, and 2) to proliferate and secrete cytokines (IFN-γ and IL-2) after stimulation with breast cancer cell lines. The following panel of cell lines was used: autologous PHA blasts, MDA-MB-468 (both Her2-negative), MCF-7 (Her2-low), Her218 and SKBR-3 (both Her2-high).
Results: Her2-CD28-zeta expressing T cells killed low and high Her2-positive breast cancer cell lines in cytotoxicity assays, where as Her2-negative T cells were not killed. Stimulation of T cells with breast cancer cell lines expressing both high and low levels of Her2 resulted in T-cell proliferation and secretion of IFN-γ and IL-2 in a HER2 dependent manner.
Discussion: We demonstrate that breast cancer cells with low levels of expression of Her2 can effectively activate T cells expressing Her2-specific chimeric T cell receptor, induce T-cell proliferation, and the production of IL-2, an important T-cell growth factor. These results indicate that T cells expressing chimeric TCRs could possibly extend the application of Her2 targeted therapies to malignancies expressing low levels of Her2.
Efficacy of Zymogen Factor VII Is Comparable to Activated Factor VII for Nonviral Gene Therapy Treatment of Hemophilia A.
Authors: Chen Steven C. , Greenberg Daniel , Ye Peiqing , Davie Earl W. , Miao Carol H. 
Published: April 2020

Abstract
A major problem for clinical treatment of hemophilia A using factor replacement therapy is the high frequency formation of inhibitory antibodies against factor VIII. This problem is also predicted to occur following strategies currently aimed at targeted genetic correction of this disease. Recombinant activated factor VII (rFVIIa, NovoSeven®, Novo Nordisk, Bagsvaerd, Denmark) has been successfully used as an effective alternative treatment for hemophilia patients who have developed inhibitors. In order to decrease the cost and fluctuation of FVIIa levels associated with frequent infusions of rFVIIa, nonviral gene transfer of factor VII (FVII) was attempted in a hemophilia A mouse model. In addition, to investigate the potential thrombotic risks associated with prolonged, high level of FVIIa expression following gene transfer, we compared the effects of gene transfer vectors encoding zymogen FVII with an engineered secreted FVIIa in hemophilia A mice. We inserted murine FVII (mFVII) cDNA into a liver-specific vector developed recently in our lab to generate pBS-HCRHPI-mFVII-A. The mFVII cDNA sequence was then modified by site-directed mutagenesis to insert a protease cleavage site in between Arg152-Ile153. The resulting construct of murine FVIIa (mFVIIa) encodes a modified protein, which can be cleaved by intracellular proteases of the furin family to secrete the activated form of mFVII. Fifty mg of either mFVII or mFVIIa the plasmid was delivered into hemophilia A mouse liver by a rapid, high volume (hydrodynamics-based) infusion method (n=8 mice/group). Phenotypic correction was evaluated using tail-clip assays by measuring the hemoglobin levels in the collected blood. This assay indicated that vectors carrying either mFVII or mFVIIa can partially correct clotting following gene transfer. Significant correction in PT and APTT was observed in mFVII or mFVIIa-treated mice. Plasma-based and platelet-based thrombin generation (TG) assays indicated that either mFVII or mFVIIa significantly shorten peak time and peak thrombin levels of TG. In addition, these effects lasted long-term in plasmid treated mice for at least three months (experimental duration). We also constructed vectors carrying human FVII (hFVII) or FVIIa (hFVIIa) to test their efficacy for correcting hemophilia A. Constructs produced supra-physiological levels (800–1200ng/ml) of hFVII or hFVIIa assayed by ELISA at days 1 and 4 post-injection. However, the levels of hFVII and hFVIIa dropped precipitously at day 14 post-injection shown by both ELISA and Western blot, most likely due to the formation of species-specific antibodies against hFVII. These results clearly demonstrate that nonviral gene therapy of either zymogene FVII or activated FVII can rescue bleeding in hemophilia A mice.
Non Invasive Screening for Fetal RHD-Genotype in All D-Negative Women Is Reliable and Cost-Effective.
Authors: Van der Schoot C. Ellen , Soussan Aicha Ait , Bonsel Gouke J. , de Haas Masja 
Published: April 2020

Abstract
About 40% of D-negative pregnant women receive antenatal anti-D despite a D-negative fetus. We have developed an automated assay for fetal genotyping using cell-free fetal DNA from maternal plasma to restrict antenatal prophylaxis to women carrying a D-positive fetus. This will save anti-D of which is worldwide shortage, and will prevent the unnecessary administration of a blood product. 1 ml of plasma is automatically presented (Tecan) to a DNA isolation-robot (Roche). The DNA-eluate is tested (after automatically pipetting) in triplicate in a real-time RHD exon7-PCR (ApplBios).
Results
Plasma from 2415 D-negative pregnant women, whose blood was sent in for 28–30th week antibody screening, has been tested. In 35 cases (1.44%) the women (10 weak D and 25 normal D-positive) were typed as D-positive by repeated serological tests, and excluded from the study.
1465 of the plasma’s (61.59%) were typed RhD positive and 915 RhD negative. Because the RHD exon7-PCR will give positive results in D-negative women carrying D-negative variant RHD genes such as RHDΨ, we determined the incidence of these genes in our studygroup. In all cases (n=39) with Ct values below 33.2 DNA was isolated from maternal leukocytes. Of the 19 cases with extremely low Ct values (&lt;32), 6 women carried an RHDΨ gene, 5 an RHD variant type VI, 3 weak D (type 1, 11 and 17) and 1 RHDdel. The 24 other women did not carry RHD genes.
To compare the plasma PCR results with cord blood serology, all women and obstetric caregivers were sent questionnaires on cord blood serology. 55% responded (n=1257). For cases with discrepant results(n=21) or incompleted questionnaires(n=31) the laboratory which performed the cord blood typing was contacted. Finally, concordant results were obtained in 1249 of the cases. In 3 cases the genotype suggests D-positivity while serology is D-negative. In 5 cases no RHD-sequences were detected in plasma but cord blood was typed D-positive. For the discrepant cases it cannot be concluded yet, which assay gave the correct phenotype.
Conclusion
This is the first large scale automated study demonstrating the feasibility of screening D-negative women to restrict antenatal anti-D to women carrying D-positive fetuses. For the recognition of women at risk for immunization, the plasma-PCR test seems to be at least as reliable as the serological test. Furthermore, postnatal prophylaxis could have been given directly after delivery in all women with a positive PCR-result, saving cord blood serology tests and possibly increasing the effectiveness of postnatal prophylaxis.
An economic evaluation demonstrated that this screening policy is cost effective in the Netherlands.
Health Care-Associated Infection (HAI) Rates Are More Pertinent Quality Indicators Than Nosocomial Infection (NI) Ones for Hematology Programs.
Authors: Chehata Sami , Grira Chiraz , Legrand Patrick , Pautas Cecile , Maury Sebastien , Kuentz Mathieu , Carlet Jean , Cordonnier Catherine 
Published: April 2020

Abstract
In hematology patients, HAI are a complex mixture of infections due to therapy, especially to transplant procedures, conditioning regimen, immunosuppressive drugs, and invasive procedures. These HAI may occur within the hospital and be related to the hospital environment (i.e., NI), or not. These different kinds of infections require very different preventive measures. The objective of our study was to assess the incidence, overlap, and causes of HAI and NI in our hematology ward over a 6 month period.
Methods and Definitions: Infections were considered to be HAI, irrespectively of their site of occurrence (hospital or not) when they were directly related to the therapeutic procedures and subsequent immune deficiency. This included: febrile neutropenia (FN) subsequent to chemotherapy, catheter-related infections, herpes virus reactivations in patients previously sero+ to the virus, primary CMV infection in CMV− patients transplanted with a CMV + donor, toxoplasmosis reactivation, invasive fungal infections, and any infection occurring within 6 months of transplant, or later in case of immunosuppressive treatment after this date. Infections were considered to be NI when they occurred ≥ 48 h after hospitalization without any symptom of infection at hospitalization. Aspergillosis was considered to be NI when there was a delay of 7 days between hospitalization and first symptoms.We retrospectively assessed 223 consecutive infectious episodes occurring between 7–12/2000 in 137 patients, including 59 stem cell transplant recipients (allogeneic: 23; autologous: 36).
Results : 204/223 infectious episodes (91,4%) were HAI while 94/223 (42%) were NI. 7/223 (3,6%) were causes of death. Among the 42 bacterial documented infections, 20 (47.6%) were due to potentially hospital-acquired strains: 16/23 (69.5%) of HAI-NIs, and 4/19 (21%) of the non NI-HAIs.
Conclusion: In a preventive infection program, the assessment of NI clearly underestimates half of the HAI in hematology patients. Although hospital environment control is of paramount importance in transplant programs, we suggest that HAI should be a better quality indicator for hematology programs than NI.
Quality of Life of Patients with Acute Myeloid Leukemia and Myelodysplatic Syndrome.
Authors: Kuehnbach Robert , Hiddemann Wolfgang 
Published: April 2020

Abstract
Background: Over the last decades, outcome from acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) has improved due to better treatment. However, little is still known about Quality of Life (QoL) of patients treated.
Methods: QoL was evaluated in patients who were randomized between April 2003 and September 2004 for treatment for AML and MDS in the German AMLCG99 trial at the university hospital of Munich-Grosshadern, Germany. QoL was evaluated using the EORTC QLQ-C30 questionnaire. During inhouse and out-patient treatment, QoL was evaluated at eight consecutive time points. Besides leukemia-specific data, duration of inhouse treatment, duration of leukocytopenia and maximal c-reactive protein (CRP max), hemoglobine levels were evaluated. QoL results were compared to already published data from normal German population and patients with small cell lung cancer.
Results: In total, 57 patients were treated, of which 76,3% had a de-novo AML, 20,3% a secondary AML and two patients an MDS. Median event-free survival was 15,4 months.
QoL data from 49 patients, who filled in a total number of 238 QLQ-C30 questionnaires, was available for analysis. Ten patients received allogeneic stem cell or bone marrow transplantation (BMT) instead of cyclic maintainance therapy.
Scores for global health status were low in the beginning, but improved significantly during treatment, finally reaching levels, that were not significantly different from normal population. No differences were seen for patients who subsequently went off protocol due to relapse or death, for karyotype or initial lactate dehydrogenases (LDH). There were neither any differences in between age groups, induction therapy groups, for patients receiving g-csf priming and those who did not or in between older patients receiving a 2nd induction chemotherapy and those who did not. Global health status did not correlate with duation of inhouse treatment, with duration of Leukocytopenia or CRP max.
Physical functioning and fatigue mainly accounted for global health status and were correlating with other functional and symptome scores. However, hemoglobin levels did not show high correlation with physical functioning or fatigue. Social functioning was significantly better for older patients (60 years and older) at the end of treatment than it was for younger ones. BMT was worsening physical and social functioning at the end of inpatient treatment, however, differences disappeared during further evaluation of QoL.
Conclusion: Rather because of than despite treatment, patient’s QoL was improving during time, partially even reaching levels of normal population.
Cardiac Toxicities after Treatment of Primary Hodgkin’s Lymphoma (HL). Analysis of the LENT-SOMA Questionnaire Registered in the Database of the German Hodgkin Study Group (GHSG).
Authors: Behringer Karolin , Elena Gilman , Anesa Lihovac , Lucia Nogova , Heinrich Seegenschmiedt M. , Volker Diehl , Andreas Engert 
Published: April 2020

Abstract
Introduction:
As a consequence of the impressive long-term remission rates in Hodgkin’s Lymphoma (HL), the reduction of treatment-related complications is becoming increasingly important. The most important treatment-related toxicities are secondary malignancies, infertility, pulmonary, thyroid and cardiac dysfunction. Here, we report results of the prospectively planned LENT-SOMA questionnaire to determine cardiac toxicities in patients successfully treated for HL.
Patients and methods:
Between 1998 and 2002, the GHSG conducted the fourth generation of clinical trials for the treatment of early, intermediate and advanced stage HL (HD 10–12) involving a total of approximately 4000 evaluable patients. The LENT-SOMA questionnaire was sent out to the participating centers 2, 5 and 10 years after randomisation of participating patients to evaluate cardiac, pulmonary, and gonadal dysfunction after HL treatment. To focus on cardiac toxicities, questions referred to cardiac risk factors, subjective and objective cardiac dysfunction, cardiac intervention and therapy.
Results:
Overall, approximately 1.900 (50%) LENT-SOMA questionnaires after 2 years and 10% LENT-SOMA questionnaires after 5 years were identified. Among all documented toxicities, oedema were most frequently reported, followed by arrythmia, cardiac failure, myocardial ischaemia, cardiomegaly, and pericardial dysfunction. The frequency of cardiac dysfunction will be evaluated according to treatment modality (chemotherapy, chemo- and radiotherapy) and it will be analysed with respect to age, gender, and cardiac risk factors. Furthermore, the amount of patients suffering from more cardiac toxicities and the probability of recovery, once a cardiac dysfunction has been detected, will be presented.
Conclusion:
Cardiac toxicities after HL treatment include oedema, cardiac failure, myocardial ischaemia, cardiomegaly, arrythmia, and pericardial dysfunction. The first analysis of the LENT-SOMA questionnaire focussing on cardiac dysfunction will be presented.
Euthanasia: History, Definitions and Clinical Guidelines from Classical Jewish Sources.
Authors: Gesundheit Benjamin , Jotkowitz Alan , Or Reuven 
Published: April 2020

Abstract
Background : End-of-life care is a central ethical problem for clinicians. The definition of euthanasia is complex and difficult causing confusion for its practical application. Historically, the term was abused leading to medical atrocities. We trust that during medical training and in continuous medical education programs ethical topics relevant for clinical practice should be addressed systematically in their cultural context. The awareness for these crucial topics will improve clinical performance.
Objective : To define philosophical concepts, to present historical events, to discuss classical attitudes in modern bioethics and to analyze classical Jewish sources which are helpful to elaborate practical guidelines for clinicians in euthanasia and end-of-life care. Reflection on these concepts by discussing classical Jewish sources might help clinicians in their judgment and process of decision-making.
Methods : A historical overview with philosophical definitions of the concepts of active versus passive euthanasia, physician assisted suicide etc are given. Sources from the classical Jewish literature are presented and analyzed also according to later traditional interpretation. Their relevance and application in modern clinical medicine is elaborated.
Results : Philosophically, the distinction between active versus passive euthanasia is crucial. Jewish sources certainly support the obligation to ease the ongoing process of dying; there is no duty to prolong the agony of the actively dying patient, which is actually prohibited. On the other hand, no practical involvement is allowed to actively induce death of the terminal patient.
Conclusion : The historical overview and the review of the literature support the need to define concepts of euthanasia for clinicians dealing with end-of-life issues, since this term caused historically and philosophically major confusion. Therefore, ethical issues should be included in formal training for physicians and other health care providers, in order to improve the clinical approach for these challenging aspects of clinical medicine. We believe, that classical Jewish sources might contribute to elaborate clinical definitions and to provide meaningful approaches for practical guidelines for clinicians.