bluuebunny/crossref_metadata_2025_split huggingface dataset:
Plasma Adrenomedullin Levels Are Elevated in Pediatric Patients with Severe Sickle Cell Anemia
Authors: Piccin Andrea , Murphy Ciaran , Eakins Elva , Murphy William , Kunde Jan , McMahon Corrina , Smith Owen 
Published: April 2020

Abstract
Nitric Oxide (NO) and endothelin-1 (ET-1) are key modulators of endothelial vessels and actively participate in the pathophysiology of vaso-occlusive events. Sickle cell anemia (SCA) is an important model for the study of blood cells/endothelium interactions. In SCA vaso-occlusive events may be severe and cumulative. Adrenomedullin (ADM) is a strong hypotension-inducer and mediates diuresis, natriuresis and vasodilation. Adrenomedullin levels are raised in chronic cardiovascular conditions. The aim of this study was to investigate the role of adrenomedullin in SCA and its relationship to endothelin-1 and nitric oxide. A cohort of 111 children with HbSS SCA was studied with 18 age-matched controls of sub-Sahara African ethnicity. The patients were in four clinical groups:
steady state (n=55, mean age 5.3 years), patients undergoing chest crisis (n=9, mean age 5.3 years), on hydroxyurea (n=30, mean age 7.2 years), on transfusion therapy (n=17, mean age 6.5 years).
A third cohort of patients with HbSC genotype (n=13, mean age 6 years) all in steady state was also included. Pro-adrenomedullin and c-terminal endothelin-1 precursor (CT-proET-1) were measured by coated tube assays according to the method by Morgenthaler et al (Clin Chem.2005;51:1823–29) and Papassotiriou et al (Clin Chem.2006;52(6):1144–51), nitric oxide was quantified according to the Griess reaction. Results (median and interquartile range) are shown in table 1. Overall pro-adrenomedullin levels were raised in chest crisis: 0.38 nmol/l (0.31–0.49) vs steady state: 0.27 nmol/l (0.25–0.32; p&lt;0.01) and control: 0.28 nmol/l (0.27–0.31; p &lt;0.01). CT-proET-1 levels were reduced in patients on hydroxyurea therapy: 43.6 pmol/l (12.6–49.6) vs control: 55.1 pmol/l (45.2–63.9; p&lt;0.05). Nitric oxide levels were significantly lower in chest crisis (19.3 mmol/l plasma; 10.7–19.9) compared to hydroxyurea (22.2 mmol/l plasma; 18.3–28.4; p&lt;0.05), and HbSC (30.6 mmol/l plasma; 20.8–39.5; p&lt;0.05) and approach significance when compared to steady state (22.5mmol/l plasma; 16.9–28.2; p=0.07). This study indicates that during chest crisis plasma adrenomedullin levels are elevated compared to normal controls and SCA patients in steady state (see figure 1). Plasma nitric oxide levels are diminished. These observations indicate that markers of ongoing vascular damage are evident in children with SCA with chest crises, and suggest that treatments that prevent onset or progression of pulmonary hypertension, such as bosentan, may also have a role in the prevention of pulmonary hypertension in children with SCA.
Table 1. ADM, ET-1, NO and hematological/biochemical values in SCA patients and controls. IR= interquartile range.
Figure 1. ADM and NO levels in sickle cell patients and controls. Bars indicate median and interquartile range. Figure 1. ADM and NO levels in sickle cell patients and controls. Bars indicate median and interquartile range.
Increased Plasma Levels of NGAL and IL-18 Correlate with Reduced GFR Providing Two Novel Biomarkers for Kidney Injury Assessment: Evidence of Early Tubuloglomerular Defect in Patients with HbS/b-Thalassemia
Authors: Papassotiriou Ioannis , Margeli Alexandra , Hantzi Eugenia , Terpos Evangelos , Voskaridou Ersi 
Published: April 2020

Abstract
Promising novel biomarkers for the assessment of the acute kidney injury (AKI) or chronic kidney injury (CKI) include plasma levels of cystatin C, Neutrophil Gelatinase-associated Lipocalin (NGAL) and Interleukin-18 (IL-18) and urinary levels of NGAL, IL-18 and Kidney Injury Molecule-1 (KIM-1). These biochemical indices have been reported to be useful for evaluating the duration and severity of kidney disease and predicting progression and adverse clinical outcomes. Furthermore, these markers have given promising results in differentiating the various causes of AKI or CKI. Progressive renal failure is one of the main complications in HbS/β-thalassemia (HbS/β-thal). Early identification of patients at high risk of developing renal failure is of great importance as it may allow specific measures to delay the progression of renal damage and thus to reduce the incidence of end-stage renal failure and mortality. Continuing our previous studies (Voskaridou et al, Kidney Int2006;69:2037–42), we explored the activation of tubuloglomerular feedback in 58 adult patients (age 42±17years, males/females 20/58) with HbS/b-thal by measuring the above mentioned parameters, along with the more classical ones such us b2-microglobulin and N-acetyl-b-D-glucosaminidase (NAG) in blood and urine using standard methodology. GFR values were calculated according to the recently proposed cystatin C–based prediction equation using only each concentration in mg/L (nephelometric assay, Dade Behring Siemens Healthcare Diagnostics): GFR [mL/min/1.73m2] = 76.7 x Cystatin C−1.18. The main results of the study were:
impairment of GFR in 21/58 patients (36%); increased plasma concentrations of NGAL and IL-18 in 34/58 (58%) and 58/58 (100%) patients, respectively; increased and/or detectable urine concentrations of NGAL and IL-18 in 54/58 (93%) and 46/58 (79%) patients, respectively; significant negative correlations between GFR and plasma NGAL and IL-18 (r=−0.735, p&lt;0.0001 and r=−0.350, p&lt;0.007, respectively) and significant positive correlations between cystatin C and urine NGAL and IL-18 (binomial, p&lt;0.005 and r=0.412, p&lt;0.03, respectively).
These findings suggest that tubuloglomerular feedback is activated in almost all studied patients with HbS/b-thalassemia. Measurements of plasma and urine NGAL and IL-18 contribute significantly to the early detection and risk stratification of renal disease in these patients. However, prior to their clinical use, these biomarkers must undergo through rigorous validation in multiple cohorts.
Hydroxyurea Therapy Increases Expression of BCAM/LU and Adhesion to Laminin in Children with Sickle Cell Disease
Authors: Johnson Clarissa E , Telen Marilyn J. 
Published: April 2020

Abstract
Vaso-occlusion is the major cause of morbidity and mortality in sickle cell disease. The tendency of red blood cells (RBCs) to adhere to extracellular matrix molecules and the vascular endothelium is believed to be a significant contributor to the vaso-occlusive process. Some published studies have shown that hydroxyurea decreases sickle (SS) RBC adhesion to some ligands, although the mechanism by which this occurs is not completely understood. SS RBCs demonstrate increased expression of several adhesion molecules, especially BCAM/LU, and also conserve functional signaling pathways that are associated with upregulation of adhesion. BCAM/LU mediates adhesion to the extracellular matrix protein laminin. We hypothesized that patients responsive to hydroxyurea (HU) therapy would exhibit reduced adhesion to laminin as well as a decrease in adhesion molecule expression. Our subjects included patients with Hb SS between the ages of 5 to 18. They were divided into three groups:
children not receiving HU therapy (n = 3); children receiving HU therapy for over 6 months (n = 5), and children initially not receiving HU but who were initiating therapy at the time of study enrollment (n = 5).
Adhesion to laminin was examined using a graduated height flow chamber to quantitate the adhesion of SS RBCs. Expression of adhesion molecules was analyzed by western blot and densitometry, using monoclonal antibody to BCAM/LU. We found that HU therapy was associated with significantly increased expression of BCAM/LU (HU: 145.8 ± 14.0 SEM; no HU: 60.8 ± 11.0 SEM densitometry units, p = .0014). This somewhat unexpected finding confirms results published earlier this year by Odievre et al. (2008). Adhesion to laminin was also increased for patients on HU (HU: 9.3 ± 5.9; no HU: .3 ± .3, p=.2), although this increase was not significant, given the variability in adhesion seen among patients and the small number of subjects. Nevertheless, the increase in adhesion corresponded to the increase in BCAM/LU expression. In contrast, adhesion to endothelial cells was decreased, although not significantly, in patients on HU (HU: 38.1 ± 38; no HU: 127.2 ± 122.5, p=.6). Our findings thus confirm earlier published data showing that HU increases the expression of BCAM/LU measured by flow cytometry and further shows that this increased expression is associated with increased adhesion to laminin but not to endothelial cells. Potential mechanisms by which HU affects adhesion molecule expression and activity merit further investigation, as does the physiologic role of these alterations. Comparison of results from patient-matched pre-treatment and post-treatment samples should also help define the effects of HU.
Figure Figure
Dermatofibrosarcoma Protuberans in 3 Patients with ADA-SCID
Authors: Kesserwan Chimene , Sokolic Robert A. , Heselmeyer Kerstin , Ortiz-Melendez Clarymar , Cohen Jeffrey , Jagadeesh Jaya , Garabedian Elizabeth , Baird Kristin , Wayne Alan S. , Cowen Edward W. , Pittaluga Stefania , Lee Chyi-Chia Richard , Sherry Richard , Bridge Julia , Finlayson Laura , Walsh Noreen , Issekutz Andrew , Candotti Fabio 
Published: April 2020

Abstract
Dermatofibrosarcoma protuberans (DFSP) is a rare malignant skin tumor with characteristic cytogenetic findings (t(17;22)(q22;q13)) resulting in a fusion COL1A-PDGFB gene. Here we report an association between DFSP and Adenosine Deaminase Deficient Severe Combined Immunodeficiency (ADA-SCID), a rare congenital immunodeficiency syndrome. We observed DFSP in 3 ADA-SCID patients. The first patient is a 3.6-year-old boy who developed a 1.5 cm soft tissue mass on the right medial thigh. The second patient is a 2.4-year-old girl with 12 plaques (0.4– 0.8 cm) distributed on the trunk, thigh and lower extremities. The third patient is a 6-year-old boy with 5 depressed plaques ranging from 1–2 cm. The diagnosis in the first two patients was confirmed by immunohistochemistry (CD34 positive/Factor XIIIa negative) and demonstration of the COL1A1-PDGFB fusion by Fluorescent In Situ Hybridization (FISH). In the third patient, lesional tissue was positive for CD34 and negative for Factor XIIIa. FISH was inconclusive and further molecular analysis (RT-PCR) is ongoing. We came across two additional ADA-SCID patients in the literature who developed DFSP and we conclude therefore that this unique and unreported association between two rare disease entities merits further study. Importantly, our observations describe the first 2 cases presenting with multiple multicentric DFSPs. DFSP may be overlooked particularly in children who may present with an indolent plaque-like lesion mistaken as birthmark or benign vascular proliferation. DFSP in ADA-SCID may be caused by an unknown virus in the setting of defective immune surveillance. The search for a viral etiology is ongoing. Alternatively, the increased DNA breakage characteristic of ADA-SCID may play a role in the onset of this malignancy.
Analysis of Chromosomal Aberrations and DNA Mutations in Bone Marrow Mesenchymal Stroma Cells from Patients with Myelodysplastic Syndrome Amd Acute Myeloid Leukemia
Authors: Blau Olga , Hofmann Wolf-Karsten , Blau Igor W. , Baldus Claudia D , Nolte Florian , Thiel Gundula , Schuemann Elke , Benlasfer Ouidad , Thiel Eckhard 
Published: April 2020

Abstract
Introduction: The biology of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) is not well understood. In MDS, ineffective hematopoiesis may result from disturbed interactions between hematopoietic cells (HC) and the hematopoietic microenvironment. Bone marrow mesenchymal stroma cells (BMSC) are key components of the hematopoietic microenvironment. BMSC from patients with hematological disorders display functional and quantitative alterations. However, the question whether BMSC in MDS/AML have cytogenetic abnormalities is discussed controversially.
Methods: In the present study, we have collected BMSC from 51 MDS and 42 AML patients at the time of initial diagnosis. Chromosome preparation was performed after cell culture for 30 days and analyzed by conventional cytogenetic (G-banding) and by different types of FISH-techniques. Furthermore, FLT3 and NPM1 mutation analysis was performed in HC and BMSC. As a control we have studied BMSC from 25 healthy individuals.
Results: Cytogenetic analysis of BMSC was successfully performed in 90 of the 93 cases. Clonal structural chromosomal aberrations, including t(1;7), t(1;10), t(1;2), t(7;9), i(1q), inv(X), del(7q), del(13q), del(17p), and others, were detectable in BMSC of 15% of patients. All cytogenetic markers were confirmed by FISH with specific probes and M-FISH. Interestingly, cytogenetic markers in BMSC differed from the aberrations in HC from the same individual. We have found cytogenetic abnormalities in BMSC from patients presenting with cytogenetic alterations in their HC as well as from those with normal karyotype. In BMSC we could not detect specific mutations of NPM1 and FLT3 (ITD and TKD), independent from the mutation status of HC. For control analysis, BMSC cultures from 25 healthy donors were prepared under the same conditions (time of culture, number and frequency of passages). BMSC from healthy donors did show normal diploid karyotypes and absence of DNA-mutations of NPM1 and FLT3.
Conclusions: Our data indicate that BMSC from MDS and AML patients are characterized by genetic instability. Lack of aberrations as detected in HC and appearance of novel clonal rearrangements in BMSC may suggest enhanced genetic susceptibility and potential involvement of BMSC in the pathogenesis of MDS and AML.
Prognostic Value of Minimal Residual Disease (MRD) Detection in Adult Acute Lymphoblastic Leukemia (ALL) – HYPER-CVAD Protocol
Authors: Tsagarakis Nikolaos , Kentrou Nektaria , Pergaminou Mirsini , Marinakis Theodore , Papadhimitriou Stefanos I , Gortzolidis Georgios , Galanopoulos Athanasios , Michali Evridiki , Papadimitriou Konstantinos , Skoumi Dimitra , Anagnostopoulos Nikolaos , Paterakis Georgios 
Published: April 2020

Abstract
Objective: HYPER-CVAD is an intensive treatment protocol of short duration, for ALL and other lymphoid neoplasms. It is consisted of 8 alternating cycles (parts A and B): in part A (cycles 1, 3, 5, 7) fractionated doses of cyclophosphamide, vincristine, doxorubicin and dexamethazone are administrated, while in part B (cycles 2, 4, 6, 8) high doses of methotrexate and aracytin. The two first cycles compose the induction therapy, while the next 6 cycles constitute the consolidation therapy, followed by two years of maintenance therapy. Protection of CNS is achieved with intradorsal injections, whereas in Ph+ ALL patients, imatinib is also administered. The aim of this study was the clinical evaluation of MRD detection in adult patients with ALL, during chemotherapy with HYPER-CVAD.
Patients/Methods: During the period 1999–2008, 30 patients were hospitalized in our hospital for ALL and were treated with HYPER-CVAD therapeutic protocol. Among them, 14/30 (46,7%) were males and 16/30 (53,3%) females (median age 43,5 years, range 16–70). Median follow-up time was 12,8 months (range 0,5–100). ALL of T-origin had 8/30 patients and of B-origin, 22/30 (1 B1-EGIL/pro-B, 17 B2-EGIL/B-common, 4 B3-EGIL/pre-B). Caryotypic analysis and FISH was done in all patients (7/30 bcr/abl+). According to classical prognostic markers of ALL: 21/30 were classified as high, 4/30 as medium and 5/30 low risk, respectively. MRD presence was detected in bone marrow samples, with flow cytometric panels, at three particular treatment time-points: completion of induction therapy (T1), completion of consolidation therapy (T2) and at the end of maintenance therapy (T3). Overall survival (OS) and disease free survival (DFS) were investigated, especially in relation to the influence of MRD presence in (OS) and (DFS), respectively. For statistical analysis, Kaplan-Meier was used.
Results: At treatment time-point (T1), MRD was detected in 10/28 (35,7%) patients, at time-point (T2) in 7/23 (30,4%) and at treatment-point (T3) in 9/17 (52,94%). The influence of MRD detection in (OS) was statistically significant (p&lt;0,05) at (T2) and (T3), while in (DFS) the detection of MRD presence in any of the standardized treatment time-points resulted in decreased DFS (T1/p&lt;0,05, T2 and T3/p&lt;0,001).
Conclusions: Our results suggest that MRD detection during therapy of adult ALL is a negative prognostic indicator for (OS) and (DFS), the independency of which has to be confirmed after prolonged follow-up time and increased number of cases.
Automated Classification of Normal and Leukemic B Cells on the ImageStream® Imaging Flow Cytometer Using Immunocytomorphological Image Characteristics
Authors: Venkatachalam Vidya , Martin Catherine Claude , Jayat-Vignoles Chantal , Faucher Jean-Luc , Feuillard Jean , Morrissey Phil , George Thaddeus 
Published: April 2020

Abstract
Leukemic B cells can be distinguished from normal B cells based on their surface staining patterns and their morphology, but the differences can be subtle and the abnormal events rare. In order to reliably differentiate normal from leukemic B cells, objective quantitation of large data sets is required. This is accomplished with the ImageStream cytometer, which automatically acquires multispectral images of thousands of individual cells in flow at very high speeds. The associated IDEAS data analysis software measures hundreds of photometric and morphometric features per cell, thereby enabling simultaneous immunophenotyping and morphology-based measurements. The features quantify individual image characteristics such as shape, size, signal strength and texture. We have leveraged these features by implementing an automated classification routine that identifies the feature combination that best separates different cell types of interest. Our automated classification routine utilizes a modification of Fisher’s Linear Discriminant criterion to measure discrimination power between two or more populations of cells. The classifier generates a composite feature consisting of a weighted linear combination of basic image features that provides maximum discrimination. Thus, our method transforms a multi-parameter classification task that can be difficult to conceptualize into a classification based on a single composite feature that can be readily conceptualized and visualized using a simple histogram output. Since the composite feature is generated from, and tested and optimized on, large numbers of cells, we obtain results that are highly objective, repeatable, statistically significant and scalable with data size. We used this classification approach to study normal and tumor cells of the hematopoietic lineage with the goal of improving diagnosis of hematological disorders. Here we show the results of using automated classification to distinguish the subtle differences between B cells of CLL patients and those of normal patients. We also used the classifier to distinguish B-cell ALL from B-cell CLL. In both cases, we used a combination of shape, size, texture and signal strength features from the SSC, brightfield and nuclear imagery generated by the ImageStream system. The data presented demonstrate the application of a novel automated cell classification method to the discrimination of normal and tumor cells within large and heterogeneous cell samples.
Genomic Modifications Occur during Maturation in Myeloid Cells
Authors: Shirasaki Ryosuke , Tashiro Haruko , Akiyama Nobu , Kawasugi Kazuo , Shirafuji Naoki 
Published: April 2020

Abstract
Objective: A chromosomal translocation plays an important role on the leukemia-generation from the normal hematopoietic cells. Various kinds of translocation-patterns and -partners have been reported; however, a typical translocation has been reported like AML-1 and MTG8 in acute myelogenous leukemia (AML) (M2), and PML and RARα in AML (M3). This implies that some common mechanism exists on the chromosomal double strand breaks and the generation of translocations. We make a hypothesis that, when normal immature cells come into a maturation process, a specific event may occur on DNA level to induce irreversible changes toward cell-death. And a target region for the irreversible change may be the near site to chromosomal translocations. We compared the digested fragment-size for restriction-endonucleases using southern blotting among hematopoietic cell lines, normal bone marrow immature progenitor cells and normal blood mature neutrophils.
Materials and Methods: Bone marrows and bloods were obtained from informed healthy persons, and mononuclear cells were prepared after Ficoll-Paque sedimentation method, and neutrophil-rich fractions were prepared with density gravity centrifugation and hypotonic RBC lyses method. Various kinds of human cell lines, cultured in DMEM with 10% FCS, were also prepared. High molecular weight DNA was extracted from these cells, and they were digested with several kinds of restriction endonucleases. After electrophoresis, the digested DNA was transferred to a nitrocellulose membrane, and southern blot-hybridization was examined. The probe used was AML-1, MTG-8, PML, and RARα, in which the 5′ and 3′ breakpoint regions observed in chromosomal translocations were selected, respectively. Results and Discussion: No changes on the blotted fragment-size were observed among in the cultured cell lines, bone marrow immature mononuclear cells, and blood mononuclear cells; however, when 5′-PML probe was hybridized after DNA was digested with HindIII, the observed blot size was changed between normal immature myeloid progenitor cells and normal blood mature neutrophilic granulocytes. This change was not due to the personal polymorphism. These observations indicate that when myeloid cells become mature to neutrophilic granulocytes, some change occurs at DNA level. PML works on the maintenance of nucleobody for cell division, and it is very important to determine whether PML gene is maintained in neutrophilic granulocytes. We now prepare a genomic library from blood neutrophils to isolate the target clone, and determine DNA sequence near at the HindIII site to identify the modification of DNA during maturation of myeloid cells.
A Case of Acute Myeloid Leukemia with Maturation (AML-M2) with C-MYC Amplification in Homogenous Staining Region (hsr) and Double Minutes (dmin)
Authors: Lee Seungok , Kim Myungshin , Lim Jihyang , Kim Yonggoo , Han Kyungja , Kim Heeje , Min Woo-Sung 
Published: April 2020

Abstract
Homogeneous staining region (hsr) and double minutes (dmin), cytogenetic indicators of gene amplification, are rare findings in hematologic malignancies, and most frequently associated with acute myeloid leukemias (AML). Herein, we report a 74-year-old female presenting AML with maturation (AML-M2) with c-MYC amplification in hsr and dmin. Conventional cytogenetic techniques showed 46, XX, hsr(4)(q25)[15]/46, XX,~20dmin[5]. The c-MYC amplification in hsr was suspected by multicolor fluorescence in situ hybridization (mFISH) using 24XCyte Human mFISH Painting Kit. Amplification of c-MYC in hsr and dmin were confirmed by fluorescence in situ hybridization (FISH) using LSI MYC(8q24) Dual color, Break Apart Rearrangement Probe. The presentation of both dmin and hsr was rare in AML cases. Majority of AML cases with c-MYC or MLL amplification in dmin or hsr were women older than 50 years, had de novo AML with complex karyotypes and had a short survival. But, this case had no chromosomal aberration besides c-MYC amplification and showed a good response to chemotherapy with a complete remission for 21 months.