bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of F8 Mutations by High Resolution Melting (HRM) Analysis.
Authors: Bignell Patricia A , Keeling David M. , Giangrande Paul 
Published: April 2020

Abstract
Approximately 50% of severe and all moderate or mild haemophilia A (HA) mutations are single base substitutions, small insertions or deletions with over 900 different mutations having been described. The usual way to screen these HA cases is by DNA sequencing of all 26 exons which is time consuming and expensive.
We have developed the technique of HRM analysis using the Corbett Rotor Gene 6000 for successful high throughput screening of HA mutations. A target sequence was first amplified in the presence of a dsDNA intercalating fluorescent dye followed by in-tube melting analysis. Initially fluorescence is high, but diminishes as the temperature is raised and DNA dissociates into single strands. The observed ‘melting’ behaviour is characteristic of a particular DNA mutation.
In our study we looked at HA mutations in 23 exons,(exons 3–26). Genomic DNA samples from 100 HA patients and carriers were amplified using primers targeting the specific exons. We detected all of the mutations by HRM analysis and each had a distinct ‘melt’ curve which enabled the carriers to be successfully screened. We detected all the HAMSTerS known mutations and our 26 novel mutations in our cohort of patients.
Our results show that HRM could be used as the initial screening method followed by DNA sequencing of the amplicon where a different ‘melt’ curve was observed. It could also be used to rescreen ‘negative’ mutation patients very quickly. This technique is simpler, cheaper and quicker than DNA sequencing method. This method may therefore be particularly useful for prenatal diagnosis and carrier diagnosis.
Molecular Identification of Individual T Cell Clones Specific for Graft- Versus-Leukemia Reaction
Authors: Foltankova Veronika , Matejkova Eva , Bartos Milan , Dendis Milos , Novotna Dana , Mayer Jiri , Michalek Jaroslav 
Published: April 2020

Abstract
Graft-verus-leukemia (GVL) effect in hematopoietic stem cell transplantation (HSCT) is usually complicated by the alloreactivity of donor T cells which leads to acute graft-versus-host (GVH) disease. GVL and GVH reactions are proved to be mediated by different T cell clones. The objective of this study was to identify and characterize T cells clones with specific antileukemia activity without mediating GVHD. We have performed primary mixed leukocyte reaction (MLR) using patient non-leukemic irradiated peripherial blood mononuclear cells (PBMC) as stimulators and donor PBMC as responders. To prepare GVL specific T cells, activated alloreactive T cells were first selectively depleted with an anti-CD25 immunotoxin (Michalek, et al. PNAS2003, 100: 1180–4). Allodepleted T cells were then stimulated in secondary MLR using irradiated leukemia cells from the same patient. Activated leukemia-reactive cells were purified by immunomagnetic selection or by FACS based on INF-γ or CD25 expression, respectively. Clonotypic assay was used for identification of individual leukemia-specific T cell clones (Michalek, et al. Lancet2003, 361: 1183–5; Michalek, et al. J Immunol2007, 178: 6789– 5). This highly sensitive assay is based on detailed analysis of T cell receptor β VDJ unique sequence (TCRB-VDJ). mRNA was extracted from sortred activated cells and cDNA synthetized by anchored reverse transcription. Target TCRB-VDJ gene sequence was amplified by anchor PCR and used to transform bacteria. Bacterial colonies were picked for plasmid isolation and subsequent direct automated sequencing of the TCRBVDJ sequences. We assume that the frequency of particular TCRB-VDJ sequences among bacterial clones after transformation are proportional to the frequency of those sequences in the original population of T cells activated by GVH or GVL reaction. We investigated the presence of individual antileukemic T cell clones in patients with acute myeloid leukemia (AML) and chronic lymphatic leukemia (CLL), and defined them by the TCRB-VDJ unique sequence. The sequences that occured in more than 10% bacterial colonies are likely to represent the most immunodominant clones. Populations of antileukemic T cell clones were oligoclonal, i.e. we observed limited number of individual immunodominat clones which plays important role in GVL reaction. In first CLL patient who had undergone HSCT, six antileukemic T cell clones were identified, four of them are considered to be immunodominant. In second CLL patient after HSCT, only one highly immunodominat autileukemic T cell clone was observed. This specific clone was further monitored by quantitative real-time PCR in patients peripherial blood.
Interleukin-17-Producing T Cells: A New Potential Player Mediating Graft Versus Host Pathology In Patients Undergoing Allogeneic Stem Cell Transplantion
Authors: Dander Erica , Balduzzi Adriana , Zappa Greta , Perseghin Paolo , Lucchini Giovanna , Todisco Elisabetta , Parma Matteo , Biagi Ettore , Di Biasio Iolanda , Villa Antonello , Mina Pamela Della , Migliavacca Maddalena , Longoni Daniela , Rovelli Attilio , Biondi Andrea , D’Amico Giovanna 
Published: April 2020

Abstract
Graft-vs-host disease (GvHD) is a major obstacle to safe allogeneic haematopoietic stem cell transplantation (HSCT), leading to significant morbidity and mortality. Recently, a subset of interleukin (IL)-17-producing cells, named Th17, have been shown to play a central role in the induction of autoimmune-tissue injuries and inflammation. The aim of our study is to analyze the Th17 population in the peripheral blood of transplanted patients experiencing (6) acute, (7) active chronic and (6) chronic GvHD. Both Elispot assay and intracellular staining showed an augmented population of Th17 (both IL-17+/IFN-g− and IL-17+/IFN-g+ producing cells) in patients with both aGVHD and active cGVHD, that can represent up to 2.4% of CD4+ T lymphocytes. In accordance, an increased IL-17 plasma level was detected both in patients with aGVHD (mean IL17A concentration= 14 pg/ml) and active cGVHD (mean IL17A concentration= 12 pg/ml), compared to healthy donors (mean IL17A concentration &lt; 4 pg/ml). Interestingly, the percentage of TH17 cells drastically decreased in patient with quiescent cGVHD. Th17 cells of patients with aGVHD and active cGVHD, were of donor origin, as demonstrated by genotyping, and, in line with what already described, showed high IL23 receptor expression (mean= 40%, range 30%–52%). Noticeably, in the liver and skin GVHD lesions, the solely CD3 cells spontaneously producing cytokines were double positive for INF-γ and IL-17A. Importantly, we demonstrated a strict correlation between the levels of TH17 cells and the clinical status of patients with GVHD. The better understanding of the role played by TH17 in the GVHD pathogenesis could open the way to new highly targeted strategies for the management of GVHD.
A Multicenter, Randomized, Dose Frequency Study of the Safety and Efficacy of Cerezyme® Infusions Every 4 Weeks Versus Every 2 Weeks in the Maintenance Therapy of Patients with Type 1 Gaucher Disease
Authors: Kishnani Priya S 
Published: April 2020

Abstract
Introduction: Cerezyme® (imiglucerase, Genzyme Corporation) is the current standard treatment for type 1 Gaucher disease (GD1). Most patients are infused every 2 weeks. A less frequent infusion schedule at the same total 4-week dose might be equally effective and more convenient.
Objective: To compare the safety and efficacy of two dosing frequencies of Cerezyme for patients with GD1.
Methods: A phase IV, multicenter, randomized 24-month trial enrolled 95 clinically stable adult GD1 patients (mean age 47 years; range 18–82 years) who had received Cerezyme ≥2 years. Patients were randomized to continue their total 4 week dose as 1 infusion every 2 weeks (Q2) or 1 infusion every 4 weeks (Q4). The primary analysis used a composite endpoint of relative change in hemoglobin level, platelet count, liver and spleen volumes, or progression of bone disease and bone crisis. A post-hoc analysis was performed based upon maintenance of therapeutic goals (Pastores et al. Sem Hematol2004; 41(4 Suppl 5):4–14).
Results: Mean 4-week doses (U/kg) of Cerezyme were 70.4±24.9 for Q2 (n=33) and 69.7±21.3 for Q4 (n=62). The primary analysis endpoint was maintained in 80.8% of Q2 and 63.2% of Q4 patients at 24 months. Per the post-hoc analysis, 100% of Q2 and 88.5% of Q4 patients maintained therapeutic goals at 24 months. No Cerezyme-related serious adverse events were reported.
Conclusions: Infusing a total 4-week dose of imiglucerase every 4 weeks appears to be safe and well-tolerated in the majority of patients in this cohort and may be considered for some stable adult GD1 patients who have achieved therapeutic goals on Cerezyme therapy. The dosing frequency of Cerezyme, as with dose itself, must be individualized based upon physician judgment and patient needs as determined by regular and comprehensive monitoring. Long term follow-up of patients on a Q4 dosing regimen is necessary to determine other clinical outcomes, such as bone mineralization and marrow infiltration.
Are Hospitals Following Guidelines for VTE Prevention? the Veneous Thromoembolism Study to Assess the Rate of Thromboprophylaxis
Authors: Amin Alpesh , Spyropoulos Alex , Dobesh Paul , Shorr Andrew , Hussein Mohamed , Mozaffari Essy , Benner Joshua 
Published: April 2020

Abstract
OBJECTIVE: The 7th conference of the American College of Chest Physicians (ACCP7) provides evidence-based guidelines on the type, dose, and duration of thromboprophylaxis in hospitalized patients at risk of venous thromboembolism (VTE), but the extent to which hospitals follow these criteria has not been well studied.
METHODS: Discharge and billing records for patients admitted to any of 16 rural and urban U.S. acute-care hospitals from January 2005 to December 2006 were obtained. Patients 18 years or older who had an inpatient stay 32 days and no apparent contraindications for thromboprophylaxis were grouped into the categories of critical care, surgery and medically ill before being assessed for additional VTE risk factors based on the diagnostic criteria outlined in ACCP7. For patients considered “at risk”, the recommended type (mechanical or pharmacologic), dose, and duration of thromboprophylaxis was identified based on the guidelines and compared to the regimen actually received, if any.
RESULTS: Among the 258,556 hospitalized patients, 68,278 (26.4%) were determined to be at risk of VTE without apparent contraindications for thromboprophylaxis. The proportions of patients who received the appropriate type and dose and duration of thromboprophylaxis were 10.5%, 9.8%, and 17.9% for critical care, medical, and surgical patients, respectively. Of those at risk, 36.8% received no thromboprophylaxis and an additional 50.2% received thromboprophylaxis deemed inappropriate for one or more reasons.
CONCLUSIONS: The implementation of ACCP7 guidelines for type, dosage, and duration of thromboprophylaxis is low in patients at risk of VTE. Fewer than 1 in 7 patients received thromboprophylaxis that met criteria for recommended type, dose and duration. The findings demonstrate a worrisome gap in the performance of evidence-based thromboprophylaxis for hospitalized patients. There is a need for physicians and health systems to improve the levels of awareness and implementation of recommended thromboprophylaxis.
Erythropoiesis Stimulating Agents (ESA) for the Treatment of Chemotherapy Induced Anemia in Patients with Hemoglobin Levels (Hb) &lt;11g/Dl- a Systematic Review and Meta-Analysis
Authors: Paladini Luciano , Clark Otavio , Clark Luciana , Engel Tobias , Faleiros Eneas 
Published: April 2020

Abstract
Background: Anemia is a frequent condition in patients receiving chemotherapy. ESA are effective to control this condition, but a recently published meta-analysis and warnings from regulatory agencies have pointed to a possible increase in mortality in these patients.
Objective: Our aim was to evaluate the safety of ESA when used according to label indications, that is, for patients with chemotherapy induced anemia (rather than cancer induced anemia) with Hb&lt;11g/dl.
Methods: We performed a systematic review and meta-analysis of all randomized controlled trials comparing the use of ESA versus placebo (or no treatment) in patients with chemotherapy induced anemia, for whom ESAs where indicated if Hb dropped below 11g/dl. The primary end-point was mortality. We searched several databases, including MEDLINE, EMBASE, LILACS and CENTRAL, among others. All suitable papers were retrived and data regarding the quality of the studies and mortality rates were extracted. Then, we performed a meta-analysis of these trials, using RevMan 5.0 software.
Results: □There were 17 studies, with 3788 patients, meeting our inclusion criteria. There were 566 deaths among the 2208 patients that received ESA and 488 among the 1580 on control groups. The meta-analysis showed no increase in mortality rates associated with the use of ESA (Relative risk (RR) = 0.95; IC 95% 0.88 to 1.03; P=0.22). Also, there was no heterogeneity in the analysis (I2=0%), showing that the results were consistent among the studies. When we analised the Hb cut-off point in 10g/dl, the meta-analysis results were similar (RR=0.97; IC 95% 0.87 to 1.07; P 0.52; I2=0%).
Conclusion: When used as indicated on label, that is, for patients with chemotherapy induced anemia with Hb&lt;11g/dl ESA are NOT associated with higher mortality rates and remain a safe option for these patients.
Preventing Infections in Cancer Patients: Antibiotics, Hematopoetic Growth Factors or Both? a Network Analysis
Authors: Herbst Christine , Naumann Frauke , Herbst Olaf , Bohlius Julia , Jüni Peter , Engert Andreas 
Published: April 2020

Abstract
Background: Infectious complications such as febrile neutropenia belong to the most serious complications of chemotherapy including stem cell transplantation for cancer. The mortality rate associated with febrile neutropenia is approximately 10%. Meta-analyses have shown that both hematological growth factors (CSFs) and antibiotics prevent infections. A comparison of these two agents or their combination has not been examined in systematic reviews. We therefore conducted a network analysis to determine the role of antibiotics and CSFs for the prophylaxis of infections in cancer patients receiving myelosuppressive chemotherapy or hematopoetic stem cell transplantation (SCT).
Methods: The network analysis included all randomized controlled trials that compared two or more of the following prophylaxis options: no prophylaxis, prophylaxis with antibiotics alone, prophylaxis using CSFs alone or prophylaxis using combinations of both, CSFs and antibiotics. Trails that did report overall survival, on study mortality, infection-related mortality, clinically documented infections, microbiologically documented infections or fever/febrile neutropenia were excluded. A Bayesian network analysis with four treatment options was performed; results are presented as odds ratios (OR) with 95% credibility intervals (CI).
Results: The best prophylaxis in the network with regard to preventing infections and infection-related mortality is the combination of antibiotics and CSFs. Comparing antibiotics alone to CSFs alone yields an OR of 1.00 (95% CI 0.69–1.52) for clinically documented infections. For the combination of both antibiotics and CSFs compared to only antibiotics or only CSFs, the incidence of clinically documented infections was also reduced (OR=0.61, 95% CI (0.46–0.77)). This is also true for the subgroup of patients receiving SCT or patients with acute leukemia.
Conclusions: For patients receiving myelosuppressive chemotherapy, antibiotics and CSFs are similarly efficient in reducing infections. The choice of agent can therefore be based on preferences of the treating physician and the adverse events expected. The best prophylactic regimen however is the prophylaxis using both, antibiotics and CSFs. The combination should be considered for patients at high risk of developing infections such as those receiving SCT or patients with acute leukemia.
Assessment of Nutrition Status in Children with Leukemia: A Study from a Developing Country
Authors: Mukhopadhyay Soma , Dutta Tusi , Dey Sonali , Datta Somnath , Mukhopadhyay Ashis 
Published: April 2020

Abstract
Background: In developing countries 40 % children suffer from mal-nutrition. It has been shown that a good nutritional status can reduce complications of treatment, strengthen the immune system and contribute to the patient’s general well being throughout treatment. A good nutritional status is therefore essential for optimal treatment of a leukemic child. The aim of our study was to see the nutritional status of leukemic children on diagnosis and effect of nutrition on outcome of therapy.
Material & Methods: During period from January 2004 to December 2007 we prospectively analyzed the nutritional status of 400 paediatric patients in Netaji Subhash Chandra Bose Cancer Research Institute, a tertiary cancer center of Eastern India. The age of the patients were 1 to 18 years (median age 12.5 years). The parameter analyzed were weight for age, height for age, total protein, serum albumin and skin fold thickness. The weight for age, height for age and skin fold thickness were taken as normal if they were between 3rd and 97th percentile curve of the growth chart recommended by the Indian Council of Medical Research. The albumin level and the total protein was considered normal if the value is equal to or more than 3gm% and 5.8gm% respectively.
Result: It was seen that total 56 patients (14% children) were low weight for age, 40 patients (10% children) were low height and 67 patients (16.75% children) had low mid arm circumference. Total 44 patients (11% children) had low serum albumin while 80 patients (20% children) had low serum protein. It was found that low weight for age, low serum albumin and low mid arm circumference were significance factors in remission induction, disease free survival and toxicity of chemotherapy (p &lt; 0.005).
Conclusion: We conclude that mal-nutrition is a major finding in cancer patients with leukemia in developing country like ours. The patient with mal-nutrition had less remission in induction, disease free survival and more toxicities during therapy as compared to well-nourished children.
A Psychometric Analysis of Parents of Children with Leukemia
Authors: Zheng Huyong , Wu Xinyi , Zhang Ruidong , Huo Liqin , Liu Huaqing , Wang Xiumei , Hu Yamei , Lu Xiaoyan , Gao Huifang 
Published: April 2020

Abstract
To investigate the anxiety and depression of parents whose children suffered from leukemia and to develop a set of theoretical guidance for later psychologic intervention, parents of leukemia children treated in Beijing Children’s Hospital (BCH) from March of 2004 to March of 2006 were evaluated with Family Environment Scale (FES) and Hospital Anxiety and Depression (HAD) questionnaires. They were tested in three phases relating to three groups: parents of newly-diagnosed-children group, parents with or without psychological intervention groups. Parents in intervention group were given individual psychotherapy, group therapy or systemic family therapy. A total of 353 questionnaires were analyzed, including 138 in newly-diagnosed-children group, 103 in intervention group and 112 in non-intervention group. Results showed:
The family environment scores of expressiveness, independence, achievement, cultural, recreational orientations and organization were significantly lower, but the scores of conflict, ethics religion and control were much higher in parents of newly-diagnosed-children group than the normative population. For example, the scores of expressiveness and conflict were 4.9 and 2.3 in normative population, but the scores were 2.3 and 3.9 in parents of leukemia children. The HAD scores of parents of newly-diagnosed-children group showed that the scores of anxiety and depression were 38.4% and 24.1% higher than the baseline value respectively. The scores of parents’ cultural, recreational orientations and independence increased to normal level in six months after their children received treatment; Fathers’ conflict score decreased to normal level in intervention group (3.0) compared to non-intervention group (4.4). The scores of parents’ anxiety and depression reduced to baseline six months after their children received treatment.
We concluded that the psychologic distress of parents was influenced tremendously by their leukemia children. At the beginning of diagnosis their anxiety, depression and conflict were significantly elevated. Most parents could cope with their leukemia children by timely and effective psychological supports. Professional psychological intervention is most helpful in reducing family conflicts.
Cdk1-Dependent Phosphorylation ofNPM Overrides G2/M Checkpoint and Increases Leukemic Blasts in Mice
Authors: Du Wei , Zhou Yun , Pike Suzette , Pang Qishen 
Published: April 2020

Abstract
An elevated level of nucleophosmin (NPM) is often found in actively proliferative cells including human tumors. To identify the regulatory role for NPM phosphorylation in proliferation and cell cycle control, a series of mutants targeting the consensus cyclin-dependent kinase (CKD) phosphorylation sites was created to mimic or abrogate either single-site or multi-site phosphorylation. Cells expressing the phosphomimetic NPM mutants showed enhanced proliferation and G2/M cell-cycle transition; whereas nonphosphorylatable mutants induced G2/M cell-cycle arrest. Simultaneous inactivation of two CKD phosphorylation sites at Ser10 and Ser70 (S10A/S70A, NPM-AA) induced phosphorylation of Cdk1 at Tyr15 (Cdc2Tyr15) and increased cytoplasmic accumulation of Cdc25C. Strikingly, stress-induced Cdk1Tyr15 and Cdc25C sequestration were completely suppressed by expression of a double phosphomimetic NPM mutant (S10E/S70E, NPM-EE). Further analysis revealed that phosphorylation of NPM at both Ser10 and Ser70 sites were required for proper interaction between Cdk1 and Cdc25C in mitotic cells. Moreover, the NPM-EE mutant directly bound to Cdc25C and prevented phosphorylation of Cdc25C at Ser216 during mitosis. Finally, NPM-EE overrided stress-induced G2/M arrest, increased peripheral-blood blasts and splenomegaly in a NOD/SCID xenograft model, and promoted leukemia development in Fanconi mouse hematopoietic stem/progenitor cells. Thus, these findings reveal a novel function of NPM on regulation of cell-cycle progression, in which Cdk1-dependent phosphorylation of NPM controls cell-cycle progression at G2/M transition through modulation of Cdc25C activity.