bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autologous Bone Marrow Cells for Treatment of Myocardial Infarction in Colombia.
Authors: Cuéllar-Ambrosi Francisco , Manuel Juán , Velásquez Oscar , Velásquez-Lopera Margarita , Navas Claudia , García Luis Fernando 
Published: October 2019

Abstract
Experimental studies in animals demonstrate the ability of the bone marrow stem cells to differentiate in cardiomyocytes, vascular endothelium cells and smooth muscle cells. There is evidence that these cells can regenerate infarcted myocardium and induce myogenesis and angiogenesis. Clinical studies in humans suggest the feasibility and safety of the utilization of the stem cells to recovery the ventricular function in patients with acute myocardial infarction. We report the first experience in Colombia using autologous bone marrow cells for cardiomyoplasty in isquemic heart disease.
This report shows the two months follow-up of four patients, three with acute myocardial infarction of the anterior myocardial wall and one patient with severe chronic isquemic heart disease. Extensive myocardial damage demonstrated by absence of viability in scintigraphic images and ejection fraction less than 40%. The patients received an optimum postinfarction medical treatment, successful coronary percutaneous intervention (three patients) or direct intramyocardial injection (one patient) to transfer of autologous bone marrow cells mobilised with granulocyte-colony stimulating factor during five days.
Demographics and results
The exercise capacity improve importantly, evidenced by increase in contractility, the six minutes test, the treadmill exercise time and the functional capacity in METS. There were not changes in the myocardial perfusion at two months follow-up, there were not complications related to the cellular transplant or the utilization of the granulocyte-colony stimulating factor. This is the first experience in Colombia with the bone marrow cells and selective intracoronary transplantation for myocardial regeneration and angiogenesis. We observed functional recovery of the left ventricle, improvement in the exercise capacity without adverse effects or complications related to the therapy.
Patient # 1 2 3 4 MI= myocardial infarction, IHD= isquemic heart disease, B/A= before/after 2 months, EF= eyection fraction, ESV=end systolic volume Age/sex 51/M 23/M 59/M 53/M Diagnosis acute MI acute MI acute MI chronic IHD Diagnosis to transplant 3 weeks 2 weeks 4 weeks > 1 year CD34 dose 19.7x10(6) 16.8x10(6) 19.5x10(6) 21.7x10(6) EF% B/A 36/43 26/40 40/41 45/55 ESV (cc) B/A 80/60 116/103 65/70 101/84 METS B/A 4/14 4/17 5/12 5/14 6-min test (mts) B/A 420/540 216/462 260/450 414/727
Characterization and Neural Differentiation of Fetal Lung Mesenchymal Stem Cells.
Authors: Han Zhong Chao , Gang Cun , Wu Feng , Jun Qing , Hong Shi , Ying Hai , Mao Zong , Liu Bin , Han Zhi Bo 
Published: October 2019

Abstract
Mesenchymal stem cells (MSCs) have been successfully isolated from a broad range of adult, fetal and other non-embryonic tissues. Fetal lung has been identified as a rich source of MSCs capable of differentiating into multilineage cells of mesenchymal origin. However, the biological characteristics and differentiation potential of fetal lung MSCs remain to be explored. In this study, we have established a series of methods for isolation and expansion of fetal lung MSCs. These MSCs could withstand 40 passages without obvious decline in proliferation ability, significant changes in morphology and expression of cell markers. Cell cycle analysis revealed that when the MSCs reached their log phase of growth, more than 90% of the cells were in G0-G1 phase while the proportion of cells in S phase and G2-M phase were about 5.56% and 2.08% cells individually. Flow cytometric analysis showed that fetal lung MSCs expressed CD13, CD29, CD44, CD90, CD105, D117, CD166 and HLA-ABC, but not CD14, CD31, CD34, CD38, CD41a, CD42b, CD45, CD49d, CD61, CD106, CD133 and HLA-DR. These MSCs could differentiate into neural cells in addition to their mesenchymal differentiation potential. Our data suggest that the fetal lung MSC population is an alternative source of stem cells for cell-based therapy of neurological defects or mesenchymal originated diseases.
Lysosomal Glycohydrolase Activities in Dendritic Cells: Is It a Function of Hematopoietic Stem Cells Differentiation Process?.
Authors: Berardi Anna C. , Manieri Pamela , Ciraci Elisa , Tribuzi Roberto , Di Girolamo Ilaria , Cavalieri Cristina , Isacchi Giancarlo , Emiliani Carla , Bottazzo Gianfranco , Orlacchio Aldo , Martino Sabata 
Published: October 2019

Abstract
A key mechanism responsible for processing of peptide-MHC class II complexes in mature Dendritic Cells (DCs) is the generalized activation of lysosomal function. Mechanisms underlie these developmental changes are controversial. Thus, it is unclear whether immature DCs can present self antigens, and which are the checkpoints that regulate antigen presentation in immature and mature DCs. Here we generated in-vitro human DCs from peripheral blood CD34+ hematopoietic stem cells (HSCs), by adding to the medium culture Flt-3, GM-CSF, IL-4, and TNF-a (cytokine cocktail, CC) at 37°C for 14 days, and analysed the lysosomal glycohydrolases production and function. Lysosomal enzymes, b-N-Acetyl-Hexosaminidase, a-Mannosidase, b-Galactosidase and b-Glucoronidase are highly increased in a wide range in DCs (14 days of culture) with respect to the CD34+HSCs. All the glycohydrolases activities measured at 3 and 7 days in-vitro culture, were similar and four times more than CD34+HSCs (day 0) respectively. Interestingly, no activities increase were observed, even when SCF, an early acting cytokine, promoting cellular proliferation, were added to the CC medium, indicating that this phenomenon is independent from the proliferation process. Moreover, LPS treatment, to induce DCs maturation, slightly enhance the specific activities of all enzymes that we tested as respect to the untreated cells. and support the evidence that the lysosomal glycohydrolases activation is up-stream to DCs maturation process. Furthermore, for the first time, this date indicated that lysosomal glycohydrolases are regulated during the stem cell differentiation process. Understanding the key mechanism leading this phenomenon is critical for therapeutic application in immunologic or neoplastic disease.
Rapid In Vivo Mapping of Hematopoietic Cis-Regulatory Elements.
Authors: Stamatoyannopouylos John A. , Hawrylycz Michael , Humbert Richard , Wallace James C. , Yu Man , McArthur Michael , Dorschner Michael O. 
Published: October 2019

Abstract
Cis-acting sequences play defining roles in the control of genes involved in hematopoiesis. However, for the vast majority of such genes, regulatory sequences remain to be defined. We describe a powerful, generic approach to identification of cis-regulatory sequences that may be applied to any gene locus in the context of any cell type. We used densely tiled primers and a novel real-time PCR-based assay to create continuous, high-resolution quantitative profiles of in vivo chromatin structure across entire gene domains. Such profiles can be analyzed using robust statistical algorithms to pinpoint disruptions in chromatin structure that are characteristic of cis-regulatory elements. We analyzed >1Mb of human genomic terrain from diverse gene loci in the context of several hematopoietic cell lines and cleanly delineated a spectrum of classical cis-regulatory activities including enhancers, promoters, insulators, and locus control regions. The approach displayed outstanding sensitivity (100%) and specificity (>99.6%) for known elements and was successful in defining novel elements even in heavily-explored terrain such as the alpha- and beta-globin loci. Since only small quantities of cells are required, the approach can be used readily in the context of hematopoietic progenitors. Systematic application of quantitative chromatin profiling to relevant genes promises to expand dramatically our understanding of the regulation of hematopoiesis.
Myelosuppresive Effects of Interferon [slpha] and Transforming Growth Factor β Can Be Reversed by Novel p38 MAPK Inhibitor SD-282 through Inhibition of Cell Cycle Arrest.
Authors: Mohindru Mani , Pahanish Perry , Collins Robert , Navas Tony , Higgins Linda , Verma Amit 
Published: October 2019

Abstract
Cytokines play important roles in the regulation of normal hematopoiesis and a balance between the actions of hematopoietic growth factors and myelosuppressive factors is required for optimal production of cells of different hematopoietic lineages. Even though the effects of Type I Interferons (IFNs α,β) and Transforming Growth Factor βs (TGF βs) as negative regulators of hematopoiesis are well documented, the exact molecular mechanisms by which such effects occur remain unknown. Our previous studies had shown that pharmacological inhibition of the p38 MAPK with commercially available inhibitors SB203580 and SB 202190 was able to reverse the myelosuppresion caused by IFN and TGF β. These inhibitors cannot be used in human studies due to toxicity and are also questioned for their selectivity in inhibiting the p38 MAPK. Thus, to confirm the role of p38 MAPK in regulating hematopoeisis, we conducted experiments with SD-282, a more potent and selective inhibitor of p38 α.
SD-282 also performs very similarly in animal and cell models to a p38 inhibitor now in the clinic. Our results show that SD-282 is able to inhibit p38 MAPK selectively in primary human erythroid progenitors (at CFU-E stage of maturation) and suppress activation of downstream kinase MapKapK-2 after IFN α stimulation. In methycellulose clonogenic assays with mobilized CD34+ cells, IFN α treatment resulted in marked suppression of both erythroid (BFU-E) and myeloid (CFU-GM) colonies, which could be reversed in the presence of p38 inhibitor SD-282. In a similar manner TGF-β2 was not able to effectively inhibit both erythroid and myeloid colonies in the presence of p38 blockade by SD-282. In further studies, we demonstrate that the primary mechanism by which the p38 MAPK pathway mediates IFN mediated hematopoietic suppression is by regulation of cell cycle progression and is unrelated to induction of apoptosis. Treatment with p38 inhibitors led to significantly lesser numbers of cells in G0/G1 phase of cell cycle arrest induced by exposure to IFN α. Altogether, these findings confirm that the p38 MAPK signalling pathway is a common effector for type I IFN and TGF beta signaling in human hematopoietic progenitors and plays a critical role in the induction of the suppressive effects of these cytokines on normal hematopoiesis. Our studies also provide a rationale for the use of SD-282 and other p38 inhibitors in cytokine mediated hematological diseases.
Berbamine-Induced Apoptosis in Human Leukemia K562 Cells.
Authors: Zhao Xiaoying , Xu Lei , Wu Dong , Xu Rongzhen 
Published: October 2019

Abstract
Purpose: To investigate apoptosis-inducing effects of Berbamine on human leukemia cells and to explore the underlying mechanism.
Materials and methods: Berbamine was dissolved in 0.9% sodium chloride to an initial concentration of 1mg/ml and subsequently diluted to desired concentrations with cell culture medium. MTT was used to examine the effect of Berbamine on cell proliferation of K562 cells. Characteristic cellular morphological changes were used as indicators of apoptosis in K562 cells while the rate of apoptosis was measured by flow cytometry assay. Expression levels of apoptosis related genes bcl-2 and bax were determined by RT-PCR and the levels of bcr/abl were evaluated by nested-PCR. Levels of Caspase 3 were measured by flow cytometry assay.
Results: Berbamine inhibited the cell proliferation significantly and in a dose-dependent manner in tested K562 cells. Its IC50 value was 5.23ug/ml. As determined by morphological observations and flow cytometry assay, Berbamine was able to induce apoptosis of K562 cells within 6 hours. The apoptosis rate of K562 was also dose-dependent. Steady-state transcript levels of bcr/abl decreased dramatically (half-quantity ratio from 1.284 to 0.506 within 72 hours following 8mg/ml Berbamine treatment. On the other hand, the protein levels of Caspase 3 surged from 18.36% to 38.25% (p<0.001) within 24 hours after treatment of 12mg/ml Berbamine. During the same period, no changes of bcl-2 or bax transcript levels were detected in the cells that were treated with 8mg/ml Berbamine.
Conclusions: Our results suggest that Berbamine is a potent inhibitor of cell proliferation and a strong inducer of apoptosis in human K562 cells. The Berbamine-induced apoptosis pathway involves down regulation of bcr/abl and up regulation of Caspase 3 expressions. Neither bcl-2 nor bax plays substantial roles in Berbamine-induced K562 cell apoptosis.
Cyclin D1 Alternative Spliced Forms Expressed in B-Cells Display Different Effects on Apoptosis and Cell Cycle.
Authors: Sola Brigitte , Gauduchon Juliette , Krieger Sophie , Roussel Mikel , Lesage Denis , Troussard Xavier 
Published: October 2019

Abstract
Cyclin D1 is an essential regulator of cell cycle progression. Over-expression of cyclin D1 is associated with tumorigenesis both in solid cancers and in hematological malignancies such as mantle cell lymphoma (MCL), hairy cell leukemia and multiple myeloma (MM). The CCND1 gene coding for cyclin D1 exhibit an A/G polymorphism at the nucleotide 870 which is responsible for the generation of an alternative splice variant called form b. The cyclin D1b is identical to the typical form a, except in the C-terminal part. The cyclin D1b has lost the PEST domain responsible for protein degradation and the threonine 286, the site of phosphorylation by the glycogen synthase kinase-3 essential for the nuclear export. Cyclin D1b has been recently described as a nuclear oncogene (Lu et al., Cancer Res63, 7056, 2003; Solomon et al., J Biol Chem278, 30339, 2003). Using real time quantitative RT-PCR, we have detected cyclin D1b mRNA in all cyclin D1a-expressing MM and MCL cell lines and in MCL patients. The cyclin D1b protein, detected with an antibody specific of the form b (a generous gift of Dr Alan Diehl) is present in a limited number of samples (1/6 MM and 1/1 MCL cell lines) and localized both in the nucleus and in the cytoplasm. We have generated B-cells conditionally (ecdysone system) expressing cyclin D1a or cyclin D1b proteins. B-cells were stimulated with ponasterone A to induce cyclin D1 synthesis and then with cycloheximide. The two alternative proteins forms displayed similar stability: calculated half-lives are 35 min and 50 min for a and b respectively. By co-immunoprecipitation and western blots experiments, we determined that, although cyclin D1b was able to bind efficiently CDK4 (to the same extent than cyclin D1a), cyclin D1b/CDK4 complexes weakly catalyzed the phosphorylation of pRB. We further showed by cytofluorometric analysis of propidium iodide-stained cells, that cyclin D1b did not allow the entry of quiescent cells within the cell cycle and did not allow the progression through the G1 phase. This implies that cyclin D1b plays a minor role in cell cycle regulation. However, cyclin D1b, such as cyclin D1a (Duquesne et al., Cell Death Different8, 51, 2001) potentiated the apoptosis induced by growth factor deprivation. It us tempting to speculate that the contribution of cyclin D1b for B-cell hemopathies does not occur through deregulation of the pRb pathway but more likely through deregulation of apoptotic pathways. We will study these aspects in the future.
Ex Vivo Expansion of Human Marrow Mesenchymal Stem Cells: A Serum-Free Medium Compared to Classical α-MEM Medium.
Authors: Meuleman Nathalie , Tondreau Tatiana , Delforge Alain , Dejeneffe Marielle , Massy Martine , Bron Dominique , Lagneaux Laurence 
Published: October 2019

Abstract
BACKGROUND: Bone Marrow mesenchymal stem cells (MSC) are pluripotent cells that have the capacity to differentiate into several tissue lineages and also the ability to support hematopoiesis. Their use in gene and cell therapy requires their in vitro expansion. Maintenance and proliferation of MSC are strongly dependent on the culture conditions such as properties of the bovine serum added in the culture medium. However, duration and culture conditions are critical for the successful clinical use of MSC.
OBJECTIVE: We have evaluated the efficiency of a commercial serum-free medium (UltraCulture, Bio-Whittaker, Walkersville, MD) supplemented with a serum substitute (Ultroser, BioSepra, Cergy-Saint-Christophe, France) in order to work without FBS and to have a more constant composition. This medium (UC) was compared to the classical medium a-MEM containing 10% FBS (Invitrogen, Merelbeke, Belgium).
METHODS: BM-mononuclear cells collected from 11 healthy donors were plated in petri dishes at a concentration of 105/cm2. After 3 days, non adherent cells were removed and culture media were added to adherent cells which were maintained at 37°C until they reached confluence. MSC expansion was analysed after the primoculture (PM) and after the first passage (P1). CFU-F (colony forming units-fibroblastic) number, phenotypic analysis and differentiation potential were also evaluated.
RESULTS::The mean culture duration was 13±2 and 8±2 days respectively for PM and P1 but the confluence was reached more rapidly for cells cultured in UC. After PM, 0.35x106 and 1.21x106 cells were obtained for MEM and UC respectively (p<0.005). Moreover, around 20% of cells cultured in MEM were CD45+ while the level of CD45+ cells was frequently < 5% in UC indicating that UC medium favored the rapid elimination of hematopoietic cells. After P1, the expansion rate was significantly higher in UC than MEM: respectively 5.13 ± 1.2 and 22.6 ± 3 (p<0.0005). The numbers of CFU-F were always higher in UC demonstrating the enhanced proliferation in serum-free medium. The phenotype of MSC was similar in the both media: SH2+, SH3+, CD44+, CD45−, CD34−, HLA-DR−. More importantly, these cells remained able to differentiate into osteocytes, chondrocytes, adipocytes and neuron-like cells in the both media confirming their multipotentiality.
CONCLUSIONS: Our data strongly support UltraCulture medium supplemented with a serum substitute as an optimal medium for MSC culture. Indeed, it allows a better cell expansion, better proliferation and preserves multipotentiality. This medium, reducing culture period and containing low concentration of serum substitute, is of major interest for clinical production of MSC.
SHP-2 Tyrosine Phosphatase Directly Controls the Proliferative Potential of Neoplasm Cells of Human Leukemia.
Authors: Xu Rongzhen , Yu Yingzi , Zhao Xiaoying , He Zhiwen , Liang Yun , Zhang Xiaohong 
Published: October 2019

Abstract
Our previous studies have indicated a stringent requirement of SHP-2 tyrosine phosphatase for normal hematopoiesis, but little is known whether SHP-2 plays a role in human leukemogenesis. Here, we show that SHP-2 plays a pivotal role in regulating the proliferating potential of human leukemia cells. We identified three types of SHP-2 proteins: m-, n- and c-SHP-2 proteins from both leukemic and normal hematopoietic cells. The m- and n-SHP-2 proteins (termed as active SHP-2) are derived from the c-SHP-2 protein (inactive SHP-2), and highly expressed on the internal membrane of rapidly proliferating cells at S/G2 phase, and in nucleus of mitotic cells at prophase and prometaphase, respectively, whereas the c-SHP-2 protein is ubiquitously expressed in the cytoplasm of both proliferating and resting cells. Intriguingly, we found that both active SHP-2 protein and its mRNA were constitutively overexpressed in leukemic blasts from various human leukemia cell lines and nearly all cases of various subtypes of human leukemia tested, relative to normal hematopoietic progenitors. Moreover, the expression level of active SHP-2 protein is positively correlated with the hyperproliferative phenotype of leukemia patients, and inversely associated with differentiation degree of hematopoietic cells. Most importantly, block of SHP-2 expression induces apoptosis and growth inhibition of leukemic clonogenic cells both in vitro and in vivo. In addition, we found that both PI3k/Akt and Ras/Erk signaling pathways are constitutively activated in human leukemias. Finally, we identified no mutation in PTPN11 among all tested leukemia patients. Based on these findings, we propose that SHP-2 tyrosine phosphatase plays an essential role in human leukemogenesis in which it may directly controls the proliferative potential of neoplastic cells of human leukemia via regulating signaling pathways involved in survival, growth and apoptosis of leukemia cells such as PI3k/Akt and Ras/Erk signaling pathways.
Implications of Gene and p53 Protein Alterations in Determining Progression of Chronic Myeloid Leukemia Phases.
Authors: Cavalcanti Geraldo Barroso , Pereira Eliane , Sheiner Marcos Antônio Mauricio , da Cunha Vasconcelos Flavia , Klumb Claudete Esteves , Maia Raquel Ciuvalschi 
Published: October 2019

Abstract
TP53 gene and p53 protein play an important role in the control of the cell cycle and DNA repair. Alterations in the TP53 gene and in the p53 protein expression have been considered an unfavorable prognostic factor for certain forms of human cancer. Mutations in the TP53 gene in patients with chronic myeloid leukemia (CML) were found in up to 30 %, specially among those in blastic crisis. At present, the only widely available technology that reliable detects and defines all mutations is DNA sequencing. However, the routine of sequencing the entire TP53 gene, in all suggestive cases of mutation, in the laboratorial routine is prohibitively costly, complex, and time consuming. To screening of those genetic alterations for p53 protein expression and TP53 gene abnormalities, in CML patients, flow cytometry (FC) and single strand conformation polymorphism of polymerase chain reaction products (PCR-SSCP) techniques are proposed. We report the results of an analysis of 72 blood samples from CML patients: 54 in chronic phase (33 in initial chronic phase (ICP) and 21 in late chronic phase (LCP)), 7 in accelerated phase (AP) and 11 in blastic crisis (BC). DNA structure for exons 5, 6, 7, 8 and 8–9 of the TP53 gene and p53 protein expression using the monoclonal antibody DO7 by FC was performed. By PCR-SSCP analysis, shift in eletrophoretic mobility of the TP53 gene, were detected in 11 patients and p53 protein were expressed in 17 out of 72 CML patients. The abnormal PCR-SSCP pattern were showed in one case of ICP (exon 7), five cases of LCP (two in the exon 7, one in the exon 8 and two in the exons 8–9), one in AP (exon 8) and four in BC (two in the exon 5, one in the exon 6 and one in the exon 8–9). In these cases, the p53 protein were expressed in seven cases (in all cases of CB and AC and 2 out of 5 LCP samples of CML patients). The statistic analysis by qui-square test showed significance between the results of the two methods (p= 0,002). These results suggest that the two concomitant methods can increase the sensibility for screening the p53 protein expression and TP53 gene abnormalities in CML patients. The presence of abnormal profile of PCR-SSCP associated to the p53 protein expression, in advanced phases of this disease, might be considered an important indicator of the phase progression of CML.