bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow Cytometric Characteristics of CD34+ Cells in Refractory Anemia: Their Diagnostic Value.
Authors: Ogata Kiyoyuki , Kishikawa Yoshifumi , Shimazaki Tomoko , Satoh Chikako , Tamura Hideto , Otsuka Yoshihiko , Dan Kazuo , Hayashi Akio , Shimazu Mitsunobu 
Published: April 2020

Abstract
(INTRODUCTION) The diagnosis of refractory anemia (RA), a subtype of myelodysplastic syndromes (MDS), is often problematic because RA often does not show cytogenetic abnormalities and dysplastic hematopoietic cells are not specific to MDS. Recent studies have suggested that the abnormalities of hematopoietic cells detected by flow cytometry (FCM) might be useful for MDS diagnosis. However, this concept has not yet been widely accepted. We reported that MDS blasts often show aberrant antigen expression. In this study, we examined the diagnostic utility of the flow cytometric characteristics of CD34+ cells in RA.
(METHODS) Whole bone marrow cells obtained from RA patients and controls (e.g., cytopenic individuals other than RA and other malignancies) were analyzed using four-color FCM. The cells were stained with CD45 and CD34, for gating CD34+ B-cell precursors (hematogones) and CD34+ myeloblasts, in combination with various other antibodies. We determined percentages of hematogones and CD34+ myeloblasts and relative mean florescence intensity (RMFI: MFI of each antigen staining ÷ MFI of isotype-matched negative control staining) of various antigens on CD34+ myeloblasts.
(RESULTS) From the data on 50 control subjects in the first subject cohort, we determined the reference range (RR) for each flow cytometric parameter. We scored data on each subject by giving 1 point to each of 10 parameters if the data of such parameters were outside RRs. These 10 parameters, which were adopted after comparing data from control and RA subjects in the first cohort, were percentages of hematogones and CD34+ myeloblasts and RMFI of CD4, CD56, CD11b, CD13, CD15, CD33, CD117 and CD133. In the first cohort, scores of all control subjects were less than 3 (40 subjects had a score of 0) and scores of 7/12 RA patients were 3 or higher. Using the same parameters and same RRs, we then prospectively analyzed the second subject cohort. Again, this approach identified 8/13 RA patients (score 3 or higher) with 100% specificity. The degree of dysplasia and presence or absence of cytogenetic abberations were not related to RA identification.
(CONCLUSION) Our method is reproducible and helpful in RA diagnosis.
Aberrant Cell Cycle and Expression Profiles of Cell Cycle Regulatory Genes in Myelodysplastic Syndrome.
Authors: Shao Zonghong , Shi Jun , Liu Hong , Sun Juan , Jia Hairong , Bai Jie , He Guangsheng , Cao Yanran , Tu Meifeng , Wang Huaquan , Xing Liming 
Published: April 2020

Abstract
Deregulation of the cell cycle machinery, especially G1/S phase deregulation, is known to contribute to the uncontrolled cell proliferation and malignant transformation in haematological malignances. However, little is known about the aberrant bone marrow cell cycle and its molecular mechanisms in myelodysplastic syndromes (MDS). In this study, we conducted the cell cycle analysis of bone marrow mononuclear cells (BMMC) before and after exposure to haematopoietic growth factors (HGFs) cocktail, in combination with the analysis of the expression of Ki-67 antigen in BMMC and CD34+ cells in twenty-seven MDS patients. In addition, we examined the expression profiles of cell cycle regulatory genes. We found that MDS patients had higher percentage of BMMC in G0/G1 phase and lower ratio of cells in S+G2/M, and higher proportion of CD34+ Ki-67+ cells. After exposure to HGFs cocktail in vitro, high proportion of G0/G1 phase cells decreased significantly in MDS. All types of four cyclins (cyclin D2, D3, E, and A1) were at higher level had high amount of mRNA transcripts in MDS, together with higher activation of CDK2, CDK6, and RB genes. Furthermore, we also demonstrated higher activation of p21 and p27 genes. These results demonstrated that G1 arrest occurred in MDS Ki-67+ hematopoietic cells. High activation of cyclins, CDKs, and RB genes are the molecular bases of large proportion of Ki-67+ proliferate BMMC population. High activation of p21 and p27 genes in MDS blocked hematopoietic cells entry into the following cell cycle.
Bone Marrow Erythroid Maturation and Cytokine Production in Refractory Anemia Patients with Positive Mitogen-Stimulated-Direct Antiglobulin Test.
Authors: Zaninoni Anna , Imperiali Francesca G. , Iurlo Alessandra , Zanella Alberto , Barcellini Wilma 
Published: April 2020

Abstract
Autoimmune phenomena, particularly directed against RBC, are described in MDS. We already reported that the amount of anti-erythroblast antibodies was significantly greater in BM cultures of MDS patients versus controls, as detected by a new method named mitogen-stimulated-direct antiglobulin test (MS-DAT). We compared BM erythroid maturation and cytokine production in patients with refractory anemia (RA) and RA with ringed sideroblasts (RARS), with or without MS-DAT positive BM cultures. MS-DAT was performed by stimulating marrow cultures with PMA and PHA and anti-erythroblast antibodies were detected by competitive solid phase ELISA. Cytokine production (IL-2, IL-4, IL-10, TNF-alpha, TGF-beta and IFN-gamma) was investigated in BM cultures by ELISA assay. Eleven out of 23 patients showed positive MS-DAT in BM (cut off value 200 ng/ml+3SD), whereas none of them was positive in PB. By comparing patients with MS-DAT positive in BM versus negative patients, we found increased erythroblast counts and peripheral signs of hemolysis (i.e higher reticulocytes, indirect bilirubin, and LDH, and lower haptoglobin) in the former, in spite of a negative MS-DAT in BP. In BM MS-DAT positive patients TNF-alpha, IL-4, and IL-10 production was lower compared with MS-DAT negative patients. BM MS-DAT positivity suggests the existence of an anti-erythroblast autoimmunity and could support steroid therapy in some early MDS patients.
Anti-erythroblast antibodies (ng/mL) Retics (%) LDH (U/L) Haptoglobin (mg/dL) Erythroblasts (%) NS PMA PHA *denotes p&lt;0.05 MS-DAT positive vs. negative MS-DAT positive 565±130* 591±113* 696±159* 1.9±0.2* 404±22* 67.7±17 44.8±13.6* MS-DAT negative 114±18 114±17 104±17 1.2±0.1 332±18 130.4±7 26.6±3.5
Megakaryocytic Progenitors in Long Term Cultures from MDS Patients.
Authors: Majado Maria Juliana , Macizo María I. , González-García Consuelo , Salido Eduardo , Molina José A. , Monserrat Jorge , Morales Alfonso 
Published: April 2020

Abstract
Introduction: bone marrow long-term cultures (LTC) have been reported to produce very poor adherent stromal layer in MDS, but there are not many studies about the behaviour of the progenitor cells in supernatant culture medium. In this study we report the results of megakaryocytic progenitors (CFU-MK) in the supernatant of LTC of patients with refractory anemia (RA).
Material and method: LTCs were performed to 11 RA patients and to 13 normal multiorgan donors as control. Bone marrow red cells were sedimented with 1% methyl-cellulose and seeded for LTC in 10ml flasks (Nalge Nunc International), with Iscover medium supplemented with horse serum, foetal calf serum, hydrocortisone and carbonic air, at a final concentration of 1x106 per ml. Flasks were placed in an incubator for 8 weeks. Half culture medium volume was renovated weekly, cell counts and assays of CFU-MK (Megacult C, Stem Cell Technologies), were performed. CFU-MK colonies were separated in three groups, containing 3–20 cells, 21–50 cells and more than 50 cells. Student t-test was used for statistical comparisons.
Results: Are in the following table, expressed as mean + standard desviation. After the third week no colony growth was observed in normal such as in MDS. Growth of CFU-MK colonies containing more than 50 cells was higher in basal control bone marrow, no statistical differences were found in the rest of the cultures.
Results CFU-MK(3–30 Cells) CFU-MK(20–50 Cells) CFU-MK (&gt;50 Cells) CFU-MK total w: week Control basal 15.75±15.79 3.33±5.42 4.88±7.44 22.42±26.43 RA basal 7.85±8.66 0.80±1.01 0.70±0.98 9.45±9.46 t-St ns ns 0.047 ns Control w 1 3.54±4.42 0.7±1.44 0.74±1.48 4.98±6.62 RA w 1 3.29±4.52 0.29±0.93 0.25±0.67 3.82±5.65 t-St ns ns ns ns Control w 2 0.55±1.14 0.07±0.24 0.05±0.25 0.67±1.43 RA w 2 0.18±0.32 0.00±0.00 0.04±0.13 0.21±0.38 t-St ns ns ns ns Control w 3 0.18±0.53 0.00±0.00 0.00±0.00 0.18±0.53 RA w 3 0.00±0.21 0.00±0.00 0.00±0.00 0.00±0.21 t-St ns ns
Discussion: No important difference was found in LTC supernatant CFU-MK in our RA patients, and this supports the idea that the stromal damage is more important, in the pathophysyology of MDS, than that of the stem cells.
Marked Elliptocytosis in Myelodysplastic Syndromes (MDS) Is Associated to Deletion of Chromosome 20q.
Authors: Baweja Mansoina , Moreno-Aspitia Alvaro , Menke David M. , Roy Vivek , Zubair Abba 
Published: April 2020

Abstract
Most common abnormalities in MDS involve the loss of chromosomal material resulting from the loss of an entire chromosome or a part of it, due to deletions or unbalanced translocations. Among partial chromosome losses, del (5q) was found to be the most common followed by del (20q). MDS can also be associated with multiple morphologic RBC abnormalities including macrocytes, ovalocytes, dacrocytes, fragmented cells and basophilic stippling. Limited cases of MDS with elliptocytosis have been reported in the literature. Elliptocytosis encompasses a heterogeneous group of erythrocyte disorders. Although acquired cases of elliptocytosis have been reported, these are most often hereditary disorders with a variable clinical presentation from asymptomatic carriers to patients with severe hemolytic anemia. This disorder is quite rare in the Unites States (US). Mutations in protein 4.1, alpha spectrin, beta spectrin, glycophorin C and band 3 have been reported in hereditary elliptocytosis. Only 7 well reported cases of marked elliptocytosis associated with MDS have been reported in the literature. Of those 7 cases, 4 of them were associated to del(20q). All of them were elderly males and all of them had ≤ 5% bone marrow blasts. Anemia is usually moderate (median 9.4 g/dL; range 8.1–12) and thrombocytopenia is mild to moderate (median platelet count 116 x 109/L; range 46–149 x 109/L). All cases were described either in Europe (n= 1; Rummens et al. Acta Haematologica1986; 75:174–177) or in Asia (n= 3; Ideguchi et al. Br J Haematol.1993, 75:387–92; Ishida et al. Cancer Genetics and Cytogenetics1997, 108:162–165; Hur et al. Clin Lab Haematology, 2004, 26: 69–72). Of these 7 cases, 3 of them were associated with protein 4.1 deficiency. To the best of our knowledge these are the first 2 cases of patients with marked elliptocytosis and MDS with del 20q abnormality reported in the US:
Patient 1 Patient 2 Age/sex 82/male 79/male FAB classification RA RA Year of diagnosis of MDS 1998 1999 WBC 3.0 x 109/L 11.5 x 109/L Platelets 82 x 109/L 79 x 109/L Hemoglobin 11.8 g/dL 10.7 g/dL Hematocrit 34.4% 31.9% Reticulocyte 4.3% NA LDH 350 u/L NA Direct Coombs Negative NA BM Cytogenetics 46 XY, del (20)(q11.2) 46 XY, del (20)(q11.2) Prior Therapy supportive care only Erythropoeitin
Evaluation for protein 4.1 deficiency in one of these 2 patients is currently pending.
The elliptocytosis noted in these MDS patients were most likely derived from acquired clones with abnormal development of the erythroid lineage and were associated with deletion of chromosome 20q. Patients with acquired elliptocytosis and associated MDS with del 20q chromosomal abnormality seem to have an indolent course with prolonged history of mild to moderate blood cytopenias and low risk to transformation to acute leukemia. Further research is needed to deduce the clinical significance of this subtype of MDS with this unusual RBC morphology
Pericyte Coverage of Abnormal Blood Vessels in Myelofibrosis Bone Marrows.
Authors: Zetterberg Eva , Vannucchi Alessandro M. , Migliaccio Anna Rita , Vainchenker William , Tulliez Micheline , Dickie Renee , Rogers Rick , Palmblad Jan 
Published: April 2020

Abstract
Neoangiogenesis, manifested as, for example high microvascular density (MVD), is an important feature of most solid tumors and also in the bone marrow of hematological malignancies. Very high MVD has been demonstrated in chronic myeloid leukemia and myelofibrosis (MF). In addition, MF marrows, as well as solid tumor vessels, display highly tortuous and branched vessels with caliber changes. However, the pathogenic mechanisms are poorly understood. Since pericyte (PC) deficiency has been described on similarly abnormal solid tumor vessels we studied vessel morphology and PC coverage in bone marrows from nine MF patients and nine controls as well as in three mouse models of MF. - In human MF marrows mean MVD was 6.2-fold higher compared to controls (P&lt;0.001) and MF vessels displayed larger mean perimeters (89±61 μm vs. 19±9 μm; P&lt;0.001). The cellular expression of VEGF-A was 6.6-fold higher in MF marrows than in controls (P&lt;0.001). Unexpectedly, we found that 99±3 % of all MF vessels were PC-coated (i.e. postive for α-smooth muscle actin) compared with only 51±20 % in controls (P&lt;0.001). To study whether these findings could be attributed to the abnormal megakaryocytopoiesis, typical for MF, we used mouse models of MF with genetic alterations affecting megakaryocyte differentiation:
low expression of the transcription factor GATA-1 (GATA-1low), over-expression of thrombopoietin, or TPO over-expressing mice lacking expression of TGF-β1 (however, fibrosis does not develop in model 3).
In all 3 models wide and morphologically aberrant vessels were detected and microvascular density was significantly higher in marrows as well as in spleens (P&lt;0.001 compared with wild type animals). All models displayed PC-coated bone marrow vessels, similar to the findings in human MF. Thus, processes leading to fibrosis, including TGF-β1, are not sufficient to deregulate angiogenesis - We conclude that human and mice MF marrow vessels display similar pathologic blood vessel architecture, with intense PC coverage, and genetic modifications of two molecules, viz GATA-1 and TPO, important for the regulation of megakaryocytopoiesis, might be of significance for this process.
Essential Thrombocythemia: Analysis of Risk Factors for Thrombotic Events in a Series of 306 Patients.
Authors: Radaelli Franca , Bramanti Stefania , Colombi Mariangela , Iurlo Alessandra , Zanella Alberto 
Published: April 2020

Abstract
Essential thrombocythemia (ET) is a chronic myeloproliferative disorder characterized by peripheral thrombocytosis and abnormal proliferation of megakariocytes in the bone marrow. Even thought thrombosis is frequently associated to ET, the risk factors of this clinical complication are still controversial. The aim of this retrospective, single institution study was to investigate clinical and laboratory characteristics associated with the occurrence of thrombotic events, with the purpose of identifying subgroups of patients who could benefit from antiaggregant and/or cytostatic treatment. 306 consecutive ET patients (109 men and 197 females, median age 58 yr) diagnosed between January 1979 and December 2002 were included in the study. At the time of analysis, 196 patients were still alive with a median follow up of 96 months. The following variables were investigated for the association with thrombotic complications: age, platelet count, previous history of thrombotic events, time from diagnosis, treatment with antiaggregant/cytostatic drugs, and cardiovascular risk factors such as arterial hypertension, obesity, hypercolesterolemia, diabetes, cigarette smoking. At the time of last follow up, 46 patients (15%) experienced at least one thrombotic event. The occurrence of thrombotic events was observed in 26/64 (40.6%) patients with previous history of thrombosis and in 20/242 (8.3%) patients with no previous history of thrombosis (p&lt;0.0001 Fisher’s exact test, odd ratio 7.6). A significant difference between the two groups of patients was also confirmed when Kaplan Meier estimates of thrombosis-free survival were compared by log-rank test (p&lt;0.0001). By logistic regression, platelet number at diagnosis did not associate with occurrence of thrombosis in the whole patient population. When patients without previous history of thrombosis were stratified according to the number of cardiovascular risk factors (none vs one vs more than one), a significant correlation with occurrence of thrombotic events was observed (Mantel-Haenszel Chi-square 5.47, p&lt;0.05). This study confirms that history of thrombosis is strongly related with risk of further thrombotic events in patients with ET, whereas platelet number at diagnosis does not seem to represent a prognostic factor. In patients with no previous history of thrombosis, the presence of other cardiovascular risk factors has to be taken into account when establishing the therapeutic approach.
Combined Muliparameter Approach to the Diagnosis of Polycythemia Vera and Essential Thrombocythemia.
Authors: Teofili Luciana , Martini Maurizio , Cenci Tonia , Larocca Luigi M. , Leone Giuseppe 
Published: April 2020

Abstract
The diagnosis of Polycythemia Vera (PV) and Essential Thrombocythemia (ET) is currently performed by excluding possible causes of secondary erythrocytosis and thrombocythosis. Recently, new typical findings of Ph-negative myeloprolipherative syndromes have been described, such as the PRV-1 RNA over-expression and the Val617Phe JAK-2 mutation. In this study we evaluated the diagnostic value of endogenous erythroid colony-assay (EECs), PRV-1 RNA over-expression and JAK-2 mutation, in the screening of patients with thrombocytosys and erythrocytosis which are candidate to a more extensive diagnostic approach including bone marrow biopsy and red cell mass evaluation. Peripheral blood samples were obtained from 56 patients referring to the hematological day hospital of our institution for erythrocythosis or thrombocythosis. EECs assay and PRV-1 expression were evaluated as prevoiusly described (Teofili L. et al, JCO 2002); Jak-2 mutation was investigated by the method described by Baxter et al. (Lancet 2005). The diagnosis of PV and ET was performed according to the PV and ET Study Group criteria. Twenty-five patients were affected by PV (19 males and 6 females, median age 68 years, range 34–80), 20 by ET, (5 males and 15 females, median age 64 years, range 31–85) 5 patients (3 males and 2 females, median age 56 years, range 35–69) and 6 patients (1 male and 5 females, median age 31 years, range 12–83) were diagnosed affected by secondary erythrocytosis and thrombocytosis, respectively. Results obtained are shown in Table 1.
In our series all PV cases were identified by means at least one positive test, whilst 20% of ET patients resulted negative to all three assays. Moreover, among PV patients, only 1 patient was positive at one single assay (JAK-2 mutation), and the vast majority of them resulted positive for two or three assays. On the contrary, among patients affected by ET, a high proportion of patients (35 %) showed only Jak-2 mutation (10%) or PRV-1 over-expression (25%).. Importantly, no false positive cases were detectable among patients with secondary erythrocytosis and thrombocythosis. In conclusion, these data confirm that PRV-1 over-expression is a feature typical not only of PV but also of ET. Nevertheless the combination of different used assays is more usefull in the differential diagnosis of erythrocythosis than of thrombocythosis.
TABLE 1 PV (%) ET (%) SE (%) ST (%) EECs pos. 18 (72) 8 (40) 0 0 PRV-1 pos. 24 (96) 14 (70) 0 0 Jak-2 mutated 17 (68) 7 (35) 0 0 Full positive 14 (56) 4 (20) 0 0 Full negative 0 4 (20) 5 (100) 6 (100)
Anagrelide Treatment and Cardiovascular Evaluation in 130 Patients with Essential Thrombocythemia (ET): Preliminary Report of the Registro Italiano Trombocitemia (RIT).
Authors: Gugliotta Luigi , Tieghi A. , Bulgarelli S. , Tortorella G. , Ciancia R. , Scalzulli P. R. , Cacciola E. , Cacciola R. , Rossi D. , Usala E. , Crugnola M. , Candoni A. , Giordano M. , Andriani A. , Santoro C. , Vimercati R. , Liberati M. , Sciorio A. , Specchia G. , Martinelli V. , Gaidano G. , Mazzucconi M. G. 
Published: April 2020

Abstract
Anagrelide treatment is considered potentially responsible of cardiovascular toxicity and side effects. For this reason, we have retrospectively evaluated 130 patients (pts) with Essential Thrombocythemia (ET) treated with Anagrelide in 10 Italian Hematological Centres belonging to the GIMEMA Group. The pts, 55 males and 75 females, had at diagnosis a mean age of 43 years (range 13–82) and a mean platelet (PLT) count of 1086x10(e)9/L. The Anagrelide treatment was started after a mean of 47 months from diagnosis(range 0–196). A previous treatment with cytoreductive drugs (mainly HU and IFN) and antiplatelet agents (mainly ASA), was performed in 65% and 98% of cases, respectively. The mean Anagrelide dose was 1 mg/day at start and 1.7 mg/day in the maintenance phase (mean PLT count below 500x10(e)9/L). Twenty-five patients (19%) discontinued the treatment after a mean of 27 months (range 1–85) for the following reasons: compliance loss (n8), no satisfactory response (n7), palpitations and tachycardia (n6), other side effects (n4), anemia (n1), renal toxicity (n1), thromboembolism (n1). During the follow-up (median 28 months) 4 thrombotic complications (1.15/100 pt-yrs) and 2 minor hemorrhages (0.57/100 pt-yrs) were registered. Moreover, 15 pts showed non lethal cardiovascular events: angina (n6), cardiomiopathy (n4), arrhythmia (n3), AMI (n2). An instrumental cardiovascular evaluation with Echocardiography (ECHO) and ECG was performed at Anagrelide start in 58 pts: the Ejection Fraction (EF) was abnormal in only one pt (EF 47%). The instrumental monitoring was done in 28 pts every 6–12 months. Four out of these 28 pts showed a decrease of the EF below the level of 50% (43%,45%,47%, and 49%), and only 1 pt discontinued Anagrelide for this reason (see table). The clinical and instrumental cardiovascular evaluation in ET patients receiving Anagrelide is becaming of routinary application and an observational prospective study of the Registro Italiano Trombocitemia (RIT) is now in progress.
ECHO EF&lt;50% IN 4 OUT OF THE 28 ET PATIENTS MONITORED DURING ANAGRELIDE TREATMENT CASES AGE EF % BASELINE EF % 1ST YEAR EF% 2ND YEAR *Anagrelide discontinuation because tachycardia and EF&lt;50% 1 44 60 45 61 2 50 50 43 50 3 51 51 49* / 4 47 47 47 /