bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transfusion Independence Achieved in a Case of Diamond Blackfan Anaemia by Essential Amino Acid Supplementation
Authors: Wright Diana , Ball Sarah , de la Fuente Josu 
Published: April 2020

Abstract
Diamond Blackfan anemia (DBA) is an inherited bone marrow failure syndrome usually presenting as pure red cell aplasia in the first few months of life, which can be associated with systemic features like short stature, skeletal, cardiac and renal abnormalities, and has an increased risk of aplasia and malignancy. The genetic defect is now known in 50% of the cases affecting ribosomal proteins and although the pathophysiological mechanism has not been elucidated, it is known that they have inefficient translation, which a recent report suggests that in some cases can be overcome by leucine supplementation both in vitro and in vivo.
We report an 8 years old boy diagnosed at 2 years of age with an Hb 3.4 g/dL requiring monthly blood transfusions. During the first year following the diagnosis he received trials of both steroids and ciclosporin, which he failed to respond to and remained transfusion dependent at the same frequency. At 3 years and 10 months of age the parents sought the advice of a nutritionist because his height was below the 2nd centile (92 cm) and weight in the 9th centile (13.9 kg). High performance liquid chromatography measurement of amino acids in blood revealed deficiency of isoleucin 40 μM (NR &gt;44) and threonine 34 μM (NR &gt;31). He received an essential aminoacid supplementation formula containing the following daily amounts: L-5-Hydroxytryptophan 0.01669 g/day, L-Arginine 0.3373 g/day, L-Histidine 0.3625 g/day, L-Isoleucine 0.6043 g/day, L-Leucine 0.6964 g/day, L-Lysine 0.3539 g/day, L-Methionine 0.5265 g/day, L-Phenylalanine 0.4628 g/day, L-Taurine 0.1669 g/day, L-Threonine 0.4908 g/day, L-Valine 0.6652 g/day, Pyridoxal-5-phosphate 0.0106 g/day, Alpha-ketoglutaric acid 0.3049 g/day. Six months after the initiation of the essential amino acid supplementation the interval between transfusions started to become longer and he became transfusion independent at 12 months. The Hb has increased steadily from 10 g/dL to 13.6 g/dL. His height has steadily improved to the 9th centile and the weight to the 25th–50th centile.
Whilst we cannot exclude that this child may have entered into spontaneous remission as it is known to happen in 5–10% of the patients with DBA, this case supports the recent report that essential amino acid supplementation may overcome the terminal differentiation defects found in the erythropoiesis of patients with DBA. A controlled trial of essential amino acid supplementation is warranted in patients with DBA.
Genetic Disruption of □pdl and □pdlr Phospholipases in Mice Leads to Impaired Platelet Adhesion and Aggregation
Authors: Wang DingYan , Reheman Adili , Connelly Phil , Wang Youdong , Maguire Graham , He Feng , Keating Armand , Ni Heyu , Wen Xiao-Yan 
Published: April 2020

Abstract
Platelet adhesion and aggregation are critical events in thrombosis. Bioactive phospholipid LPA (lysophosphatidic acid) has been identified as an important agonist for platelet aggregation. Plasma LPA can be generated from phospholipid substrates by phospholipase A1 (PLA1) and phospholipase D (PLD). We previously identified two novel PLA1 enzymes, designated lpdl (lpd lipase) and lpdlr (lpdl related lipase). Together with phosphatidylserine phospholipase A1 (PS-PLA1), these three phospholipases form a unique PLA1 lipase subfamily. Phospholipids are important structural components of cellular membranes. Recent studies demonstrate that lipid raft microdomains on the platelet membrane contribute to platelet activation. The gathering of membrane lipid rafts is necessary for ADP-mediated platelet activation. Disruption of lipid raft results in a reduction of ADP-induced platelet aggregation. In addition, it is reported that alteration of membrane lipid composition also affects the function of platelet β3 integrin (GPIIbIIIa). Thus, phospholipases may play an important role in platelet function. However, the roles of lpdl and lpdlr in platelet activation and thrombosis are unknown. To study the function of lpdl and lpdlr, we have recently knocked out both lpdl and lpdlr genes in mice by deleting their exons I and II, including their translation start codon ATG. RT-PCR confirmed that neither lpdl nor lpdlr gene is expressed in homozygous lpdl−/− or lpdlr−/− mice but expressed in wild-type (WT) tissues, indicating successful knockout of these phospholipases. Next, optical platelet aggregometry was used to assess platelet aggregation in platelet rich plasma (PRP). Platelet aggregation was induced with 5, 10 and 20 μM ADP in pooled PRP (3×108 platelets/mL) from age- and sex-matched WT, heterozygous and homozygous mice. Our results demonstrated that both the lpdl−/− and lpdlr−/− mice have decreased platelet aggregation after ADP stimulation. We further studied thrombus formation in lpdl−/− mice using an ex vivo perfusion chamber model on collagen type I-coated rectangular glass microcapillary tubes. Our preliminary data showed that both platelet adhesion and aggregation were impaired in the lpdl−/− mice. We are repeating these experiments and are examining lpdlr−/− mice using the same perfusion chamber model. In vivo thrombosis models with intravital microscopy and the mechanisms of how lpdl and lpdlr enzymes regulate platelet activation will also be investigated. These studies could potentially identify a novel pathway for regulation of platelet function, which may lead to development of new diagnostic and/or therapeutic methods for atherothrombosis. (Drs. D. Wang and A. Reheman contribute equally to this work)
ATG Plus Cyclosporine-a Should Be Used as Immunosuppressive Therapy in Patients with Severe Aplastic Anemia-Systematic Review and Meta-Analysis
Authors: Gafter-Gvili Anat , Ram Ron , Gurion Ronit , Paul Mical , Yeshurun Moshe , Raanani Pia , Shpilberg Ofer 
Published: April 2020

Abstract
Background: Immunosuppressive therapy (IST) is the therapeutic alternative for patients ineligible for allogeneic transplant. Although the combination of anti-thymocyte globulin (ATG) and cyclosporine-A (CsA) is considered the “gold standard” for these patients, the essentiality of using both drugs can be challenged.
Objectives: This study aims to compare between treatment with ATG + CsA and ATG alone for patients with severe aplastic anemia (SAA), and with non-severe aplastic anemia (NSAA).
Methods: Systematic review and meta-analysis of randomized controlled trials of patients with aplastic anemia comparing ATG and CsA with ATG alone. The Cochrane Library, MEDLINE, conference proceedings and references were searched until 2008. Outcomes assessed were: all-cause mortality, overall hematological failure, refractory disease. Relative risks (RR) with 95% confidence intervals (CIs) were estimated and pooled.
Results: Our search yielded 4 trials. For patients with SAA there was a significant reduction in all-cause mortality at 3 months in the ATG+CsA arm compared with the ATG alone arm (RR 0.50 [95%CI 0.29–0.85]). This effect was also shown at 1 year (RR 0.54 [95%CI 0.30–0.99]) (Figure) and at 5 years (RR 0.58 [95%CI 0.36–0.93]). There was also a reduction in overall hematological failure (RR 0.67 [95% CI 0.49–0.90]) and in the number of patients with refractory disease (RR 0.51 [95%CI 0.33–0.81]) in the ATG+CsA arm.
In patients with NSAA, there was no difference in mortality at 6 months (RR 1.03 [95%CI 0.07–15.78]) and at 5 years (RR 1.03 [95%CI 0.07–15.78]) or in refractory disease (RR 0.89 [95% CI 0.40–1.99]), when ATG+CsA was compared to ATG alone. However, there was a reduction in overall hematological failure in the ATG+CsA arm (RR 0.70 [95% CI 0.42–0.88]).
Conclusions: Our review demonstrates that the combination of ATG with CsA significantly reduces short and long term mortality in patients with severe but not with non severe aplastic anemia. The combination of both drugs should therefore be considered the gold standard only for patients with SAA.
Figure Figure
Evaluation of Pegfilgrastim Efficacy in Peripheral Blood Stem Cell Mobilization of Patients with Lymphoma
Authors: Bouabdallah Krimo , Bijou Fontanet , Dilhuydy Marie-Sarah , Banos Anne , Milpied Noel-Jean 
Published: April 2020

Abstract
Twenty four patients (pts) with planned autologous stem cell transplantation for lymphoma diseases (Hodgkin’s disease=4; non-Hodgkin’s lymphoma=20) received chemotherapy (CT) (Induction CT=3 and salvage regimen= 21) followed by a fixed single dose (6 mg) administration of Pegfilgrastim (PF) after the last day of CT for peripheral blood stem cell collection (PBSC) (target cell dose of 3 2×106 CD34+/kg). Median age was 53 yrs (24–68) and median weight was 72, 5 kg (45–98). Among the 24 pts, 7 received more than 2 lines of CT regimens. The injection of PF was well tolerated. Median time interval between day 1(D1) of the cycle of CT mobilization and first leukapheresis session was 14 days (10–18) while the median time interval between injection of PF and first leukapheresis session was 9 days (6–13). Stem cell collection was started when the absolute number of circulating CD34+ cells was &gt;10×106/L and performed with standard volume leukapheresis. Median CD34+ cells level at D1 of leukapheresis was 35, 5/mm3 (11–320) and interestingly, more than 35 % of pts could reach this median level of CD34+ early after PF injection (around D6). Notably, 22 pts reached the target cell dose in 2 sessions of leukapheresis or less (10 pts after 1 session, 10 other pts after 2 sessions, 2 pts after 3 and 4 sessions respectively). The median number of leukapheresis sessions was 2(1–4) and the median CD34+ cells harvested was 4×106/kg (0,8–26,6). Two pts (DLBCL = 1 and FL = 1) could not reach the level of CD34+ required to start leukapheresis and both became secondary refractory to CT. In univariate analysis, PBSC collection of &gt; 4×106/kg was highly correlated with pts who started their collection at D9 of PF administration (P=0,01) and with those presenting a CD34+ cells level &gt; 35.5/mm3 at D1 of leukapheresis (P=0,033). White blood cells level higher than 9 G/l was also predictive of circulating CD34+ cells &gt;35,5/mm3 (P=0,033). These data suggest that PF may represent an attractive option for PBSC mobilization particularly for pts with lymphoma when optimal compliance of frequent sequential regimens of CT is required. We also emphasize that stem cell mobilization is effective even in pts in second or subsequent salvage CT regimen. Importantly, the circulating CD34+ count should be performed from D6 of PF administration. The presentation will include the updated data.
The Philadelphia Chromosome Is Not a Stable Structure and Is Vulnerable to Further Rearrangements during Disease Progression in CML
Authors: Virgili Anna , Brazma Diana , Chanalaris Anastasios , Grace Colin , Nacheva Elisabeth 
Published: April 2020

Abstract
Chronic myeloid leukaemia (CML) is a pluripotent haematopoietic stem cell disorder characterized by the expression of the BCR/ABL1 fusion gene, which commonly results from formation of the Philadelphia chromosome (Ph) after a t(9;22)(q34;q11) or related variant rearrangement. BCR/ABL1 is a constitutively activated tyrosine kinase and its amplification has been described in association with resistance to imatinib in CML patients. BAC array CGH analysis on CML patients and CML cell lines (Brazma et al., 2007) revealed unexpected genomic imbalances in form of duplications and high copy number gains affecting the region immediately downstream of the ABL1 gene at the Philadelphia (Ph) chromosome in patients at the blast crisis stage. We aimed to confirm and map these amplifications by fluorescence in situ hybridization (FISH) on 19 CML patients in accelerated phase/blast crisis and 10 CML cell lines (KU812, K562, MEG-01, MC3, BV173, EM-2, LAMA-84, KCL-22, JK-1 and CML-T1) with more than 1 copy of the BCR/ABL1 fusion gene. We used a range of BAC probes and 9q and 22q sub-telomeric probes in order to do the FISH mapping. While the majority of the analysed patients and cell lines (12/19 patients and 6/10 CML cell lines) had 2 identical Ph chromosomes, 2 main groups of abnormalities were identified. Firstly, gains of the Ph chromosome taking the form of ider(22)t(9;22) chromosome were detected in 1 or more copies in a subset of bone marrow cells of 5/19 patients and, secondly, high copy number gains were seen in 2/19 patients and 2/10 cell lines (K562 and KU812). The amplified region was variable in size spanning from 400 Kb up to 1.5 Mb downstream of the ABL1 gene. In 1 patient, 7 different cell sub-clones harbouring increasing levels of amplification were found. The gains resulted in formation of different chromosome structures-from an ider(22)t(9;22) to markers with tandem amplifications, which included the BCR/ABL1 fusion with variable in length sequences downstream of the ABL1. Duplication of some 571 Kb downstream of ABL1 was also detected in 1 of the 2 apparently normal Ph chromosomes in the MC3 cell line, while a larger duplication (5.16 Mb) was found in another cell line (MEG-01). These findings confirm the presence of duplications and high level amplifications at the der(22) t(9;22) in CML patients and that the sequences involved are variable in length, indicating that the Ph chromosome is an unstable structure and vulnerable to further rearrangements during disease progression.
Ikaros Is a Frequently Affected Hematopoietic Differentiation Factor in Pediatric Relapse-Prone Precursor B-Cell Acute Lymphoblastic Leukemia
Authors: Kuiper Roland P. , van Leeuwen Frank N. , van der Velden Vincent , van Reijmersdal Simon V. , de Vries Jeltje , Keijzers-Vloet Suzanne T.M. , Hehir-Kwa Jayne , van Wering E. , van Dongen Jacques JM , Hoogerbrugge Peter M. , van Kessel Ad Geurts 
Published: April 2020

Abstract
Relapse is the most common cause of treatment failure in childhood acute lymphoblastic leukemia (ALL), and is difficult to predict in the majority of cases. Here, we performed genome-wide copy number profiling of 34 paired diagnosis-relapse samples from children diagnosed with precursor-B cell ALL. The majority of the copy number abnormalities were preserved between matched diagnosis and relapse samples, but lesions unique in either of the two samples were observed in 82% of the cases. In 68% of the cases lesions present at diagnosis were no longer detected in relapse samples (including recurrent lesions affecting the PAX5, CDKN2A, and EBF genes), indicating that these lesions were secondary events, absent in the original therapy-resistant progenitor clone. Deletions in the IKZF1 gene encoding the hematopoietic differentiation factor Ikaros were observed in 38% of the diagnosis samples, which is &gt;6-fold the amount detected in an unselected group of pediatric ALLs. Tiling-resolution oligo arrays were used to map the breakpoints, which demonstrated that the protein-coding exons 3–6, encoding the DNA-binding Zn-finger domains, were most commonly deleted. Sequence analysis revealed that point mutations in IKZF1 do occur but are less frequent. Furthermore, IKZF1 deletions were always preserved in relapse. Together, we conclude that IKZF1 deletions are frequent events in therapy-resistant clones of relapse-prone pediatric precursor B-ALL.
Circulating Angiogenic Cytokines in Chronic Lymphocytic Leukemia: Relationship to Modern Prognostic Factors and Effect of Fludarabine-Based Treatment
Authors: Smolej Lukas , Andrys Ctirad , Pekova Sona , Kucerova Lenka , Hrudkova Monika , Belada David , Maly Jaroslav 
Published: April 2020

Abstract
Angiogenesis has been studied in chronic lymphocytic leukemia (CLL) since mid 1990s. Given the extraordinary heterogeneity of clinical course of CLL patients, angiogenesis assessment may potentially improve prognostic stratification. There are numerous reports on increased markers of angiogenesis in CLL; however, data on possible association of angiogenic markers with modern prognostic factors are very scarce. Therefore, we evaluated prognostic significance of angiogenic cytokines, namely basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), endostatin, and angiopoietin-2 (Ang-2) in patients with untreated CLL. Plasma levels of angiogenic factors were quantified using enzyme-linked immunosorbent assay (bFGF, VEGF, n=106; endostatin, n=83, Ang-2, n=55). Eighty healthy donors served as control group. Relationship of angiogenic markers to modern prognostic factors (IgVH mutation status, cytogenetic aberrations detected by fluorescent in situ hybridization [FISH]) was also analyzed. Results: bFGF, VEGF, and endostatin were significantly elevated in CLL patients when compared to controls (bFGF, VEGF: p&lt;0.0001, endostatin: p=0.039); Ang-2 levels were not different from the control (p=0.192). There were no differences in angiogenic factors with regard to gender, clinical course or modified Rai stage. Similarly, no association of angiogenic factors with IgVH or FISH was identified. Interestingly, in a pilot subgroup of 16 patients with serial measurements before and after successful fludarabine-based combination treatment, levels of bFGF as well as VEGF decreased significantly (p&lt;0.0001 and p=0.0027, respectively). In conclusion, our study confirms that angiogenic signaling is intensive in CLL. Fludarabine-based treatment may have intrinsic antiangiogenic properties. Further research is needed for better understanding of angiogenic processes, especially with respect to possible refinement of individual patient’s prognosis in this disease.
Prognostic Value of β2-Microglobulin (β2M) Expression on Lymphocyte Surface in Chronic B-Lymphocytic Leukemia (B-CLL)
Authors: Volkova Maya , Molodyk Alvina A. , Bialik Tatiana E. , Frenkel Marina A. 
Published: April 2020

Abstract
Background: Modern therapy diminished prognostic significance of serum β2M level.
Aims: This study was designed to investigate the correlation between the number of lymphocytes expressing β2M, serum β2M level, and the results of treatment.
Methods: 45 patients (pts) were included (Binet stage B, 36 and C, 9). β2M expression on the lymphocyte surface was detected with immunocytochemical method by means of Dako-Cytomation LSAB2 System HRP, β2M serum level by ELISA.
Results: In 29 pts β2M was expressed on 80–100% of lymphocytes (group I), in 16 pts on 35–79% (group II). Number of lymphocytes expressing β2M did not correlate with lumphocyte dimension, form of nucleus, and presence of prolymphocytes in peripheral blood (PB). β2M serum level ranged from 3,0 to 10,5 mg/L. Number of leukocytes and lymphocytes in PB and bone marrow in both groups had no difference. Number of pts with normal β2M serum level (3,5 mg/L or less) was practically the same (group I, 13,8%; group II, 18,8%). Among pts with high β2M serum level (&gt; 6 mg/L) there were 28% in group I and 69,0% in group II. Increased level of LDG (&gt;450 MU/L) was detected in 10% of pts of group I and in 50% in group II. Treatment in both groups was similar: regimens with fludarabine–gr.I, 23 of 29 pts and gr.II, 12 of 16; R-COP–gr.I, 3 pts and gr.II, 2 pts. Two pts in gr.II received leukeran; 3 pts in gr.I didn’t need any treatment. Of 42 pts treated 20 pts achieved complete remission; 13, partial remission, and 9 pts failed. In gr.I complete remission was achieved in 17 of 26 treated pts (65,4%), in gr.II, in 3 of 16 (18,8%), p=0,0038. Partial remission was received in 6 pts of gr.I (23,1%) and in 7 pts (43,8%) of gr.II, p=0,14. Three pts of gr.I (11,5%) and 6 pts of gr.II (37,2%) failed (p=0,05).
Conclusion: Comparison of the therapy results showed that β2M expression on large number (80–100%) of lymphocytes in B-CLL can be considered as the favourable prognostic sign. It was associated with lower level (&lt;6 mg/L) of serum β2M (in 71,2% vs. 31% of pts, p=0,009) and LDG (90% vs. 50% of pts, p=0,0051). Disclosure: No relevant conflicts of interest to declare.
Serum Folate Measurement by Chemiluminescence in Patients with Paraproteinaemia
Authors: Wakeman Lisa J , Lewis John O , Morris Keith , Benton Ann , Munro Roger C , Al-Ismail Saad 
Published: April 2020

Abstract
Introduction: Paraproteins cause interference in many assays systems due to increased viscosity and/or by non-specific binding to either analytes or reagents which may variably affect the results. There is abundant evidence to indicate that performance characteristics of automated, competitive protein-binding assays for folate assays are normally sound, but there is a paucity of data comparing folate assays in patients with paraproteinaemia compared to normals.
Method: Serum samples were prepared from venous blood taken into Greiner Vacuette containers (code: 456018) in fifty patients with paraproteinaemia (IgG Kappa, n = 20: IgG Lambda, n = 9: IgM Kappa, n = 6: IgM Lambda, n = 4: IgA Kappa, n = 5: IgA Lambda, n = 3: multiclonal, n = 3). Serum folate assays were measured according to manufacturer’s instructions on Beckman-Coulter Access and Abbott Architect haematinics analysers on the day of sample collection.
Results: The ratio of means of serum protein concentrations in the paraprotein sub-groups compared to the mean values of age/gender related reference ranges were: IgG = 1.57: IgM = 7.8: IgA = 6.3. Serum albumin concentration was normal in all patients. Mean serum folate results (±1SD) for paraprotein patients and normals controls (laboratory personnel) are shown in the table below:
Serum Folate (μg/L) Access Architect paired t-test Normal controls (n = 50) 5.98 (±3.57) 5.01 (±3.14) p&lt;0.05 Paraprotein patients (n = 50) 5.08 (±2.79) 5.01 (±3.03) p0.700
When analysed using the paired-t-test serum folate was significantly different in normal subjects using the Access compared to the Architect. However, no such significant difference between the two systems was observed in the paraproteinaemia group. Although the mean serum folate in normals was higher compared to the paraprotein group as analysed by the Access, this difference was not significant when analysed using the two-sample t-test. Interestingly the mean serum folate when analysed in the two groups using the Architect gave identical mean values. There were no statistically significant differences between the two methods in any of the paraprotein sub-types (p&gt;0.05 in each case). Conclusions: The complex and variable mixture of proteins present in paraproteinaemias pose potential erroneous measurement in immunoassay systems. This is because they may interfere with the associated antigen-antibody reactions which are dependant on complimentary matching shapes being assumed by the antibody variable regions of the immunoglobulin. Despite this, our data using two distinct chemiluminescence analysers for folate measurement indicate no statistically significant difference for this parameter between patients with paraproteinaemia compared to normal controls. However, it has yet to be determined whether differing results between these two groups are generated using different analytical techniques.