bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunohistologic Identification and Quantification of Human Bone Marrow Hematopoietic Stem Cells.
Authors: Quaini Federico , Lazzaretti Mirca , Carranza Lorena , Ferraro Francesca , Bonomini Sabrina , Urbanek Konrad , Mancini Cristina , Giuliani Nicola , Anversa Piero , Rizzoli Vittorio 
Published: April 2020

Abstract
A recent experimental and clinical observation on the use of bone marrow-derived stem/progenitor cells (BM-SC) to treat non-hematologic degenerative diseases has renovated the long-standing knowledge of their biology and function. However, the paradoxical lack of histological studies limits the identification of SC within the human BM, constraining their purification and further obscuring the sites where SC fate is decided. In an attempt to identify human hematopoietic BM-SC and their relationships with the tissue environment, bone marrow biopsies from 31 subjects without BM involvement by hemo-lymphoproliferative diseases were analysed with stringent morphometric criteria for the quantification of cells expressing CD117 (c-kit) and CD34, as candidate SC markers, and/or CD45, CD3, CD20, myeloperoxidase (MPOX), CD68, Glycophorin A (GlyA) and Factor VIII (vWF), as lineage commitment. Immunofluorescence combined with confocal examination was also performed to allow the simultaneous detection of multiple surface/cytoplasmic antigens together with transcription factors. Hematopoietic cells constituted 54% of the tissue among which MPOX, GlyA and vWF positive cells represented 43.8%, 13.5% and 0.06% of marrow cells, respectively. The frequency of CD34pos cells was 93.5±43.6 each 105 total nucleated cells; c-kit involved 2.7±2.6/105 cells and c-kit-CD34 co-expression was present in 1.14±0.98/105cells. When the incidence of CD45 was evaluated among these two cell populations only 15% of c-kitpos and 49% of CD34pos cells showed hematopoietic lineage commitment, indicating the presence in the human bone marrow of undifferentiated SC. The analysis of the expression of Terminal Deoxynucleotidyl Transferase (TdT) in hematopoietic cells demonstrated that 10% of c-kitpos, 20% of CD34pos and 0.62% of CD45pos cells possessed the nuclear transcription factor. The distribution of BM-SC was predominantly scattered throughout the tissue although interstitial and paratrabecular clusters were detected and are currently under intense scrutiny in search of BM-SC sites of activation. In a subset of patients, the histological analysis was complemented by the cytofluorimetric estimation of the same BM cells populations. The results of this study may contribute to the improvement in the isolation and characterization of BM-SC aimed to the identification of their niches and to an appropriate clinical application in the field of regenerative medicine.
Abnormal Pulmonary Function in Adults and Children with Sickle Cell Disease.
Authors: Morris Claudia R. , Gardner Jennifer , Wen Andrew , Robertson Shanda , Hagar Ward , Vichinsky Elliott 
Published: April 2020

Abstract
Introduction: Chronic lung disease is a fatal complication in sickle cell disease (SCD) often undiagnosed until late stages. Early detection and treatment of risk factors may improve survival. Since asthma, pulmonary hypertension (PHT), and acute chest syndrome (ACS) are potential risk factors, in 1998, we began screening patients with history of ACS or wheezing with pulmonary function tests (PFT) and echocardiogram (echo).
Methods: A chart review was completed on the 362 adults and children evaluating history of asthma, lung function (PFT) and PHT (echo, tricuspid regurgitant jet velocity ≥ 2.5).
Results:
PFT Findings All Ages Adults Children PHT (All Ages) Study performed 34% (124/362) 41% (95/232) 22% (29/130) Echo: 90% (111/124) Abnormal 85% (106/124) 87% (83/95) 79% (23/29) 51% (54/106) Obstructive (OBS) only 31% (33/106) 31% (26/83) 30% (7/23) 42% (14/33) Restrictive (REST) only 30% (32/106) 28% (23/83) 39%(9/23) 38% (12/32) Both OBS + REST pattern 27% (29/106) 29% (24/83) 22% (5/23) 69% (20/29) “Abnormal,” not specified 11% (12/106) 12% (10/83) 9% (2/23) 67% (8/12) DLCO &lt; 65% predicted 48% (45/94) 53% (42/80) 21% (3/14) 54% (26/48) Diagnosis & Treatment Clinical asthma diagnosis 13% (47/362) 14% (33/232) 11% (14/130) 43% (20/47) Receiving asthma treatment 51% (24/47) 45% (15/33) 64% (9/14) 50% (12/24) Bronchodilator only 23% (11/47) 24% (8/33) 21% (3/14) 45% (5/11) Inhaled steroid 23% (11/47) 18% (6/33) 36% (5/14) 55% (6/11) Singulair 4% (2/47) 3% (1/33) 7% (1/14) 50% (1/2) PFT suggests asthma 50% (62/124) 53% (50/95) 41% (12/29) 53% (33/62) +PFT, + asthma treatment 16% (10/62) 14% (7/50) 25% (3/12) 80% (8/10)
One hundred twenty-four patients underwent PFTs, of which 111 (85%) were abnormal. Obstructive and/or restrictive disease with abnormal diffusion capacity were widespread. Of patients with an obstructive pattern, only 16% were receiving any asthma treatment. Forty-seven patients (33 adults and 14 children) were diagnosed with asthma. However, only half were receiving any treatment: 23% bronchodilators, and 23% inhaled steroids. Echo was performed on 90% of patients with PFT data. Half of all patients with abnormal PFTs, and 69% of those with obstructive/restrictive patterns, had PHT.
Conclusion: While a more severe population may have been tested, this data suggests abnormal lung function is prevalent in SCD and is associated with PHT. Recent data suggests abnormal NO metabolism may link asthma and PHT in SCD. In conclusion, our data suggests patient morbidity will be decreased by regular screening with PFT and echo followed by early treatment for asthma and PHT.
Lactadherin Enhances the Adhesion of Sickle Cells to Vascular Endothelial Cells.
Authors: Guchhait Prasenjit , Dasgupta Swapan K. , Lopez Jose A. , Thiagarajan Perumal 
Published: April 2020

Abstract
Adherence of red blood cells to the endothelium initiates vaso-occlusion in sickle cell anemia. The increased adhesiveness of sickle erythrocytes is accompanied by several changes in the lipids of the erythrocyte membrane, including increased expression of phosphatidylserine (PS). One important PS-binding protein is lactadherin (also known as milk fat globule-EGF factor 8), a 45-kDa glycoprotein containing an Arg-Gly-Asp (RGD) sequence. Lactadherin is secreted by many cell types, including macrophages, and is known to have a role in the clearance of apoptotic lymphocytes by binding PS on the cell surface and thus presenting a binding site for macrophage integrins, including αvβ3. We recently showed that lactadherin, added exogenously to both normal and sickle erythrocytes, enhances phagocytosis of these cells by macrophages. Here, we investigated the effect of bovine lactadherin on the adhesion of sickle erythrocytes to the endothelium in flowing blood. Lactadherin (100 nM) promoted the adhesion of sickle erythrocytes to histamine-stimulated human umbilical vein endothelial cells (HUVECs) at shear stresses from 2.5 to 10 dyne/cm2. Adhesion was inhibited significantly by abciximab (10μ g/ml), an antibody against αvβ3 (P&lt;0.05). Normal erythrocytes were induced to adhere to stimulated HUVEC in a lactadherin-dependent manner by treatment with 10 mM N-ethylmaleamide and 4 μ M calcium ionophore A23187—treatment that exposes phosphatidylserine on the erythrocyte surface. These results indicate that lactadherin mediates sickle cell adhesion to the endothelium by bridging PS on erythrocytes to αvβ3 integrin on the endothelium. We propose that sickle erythrocytes acquire lactadherin as they traverse macrophage-rich zones such as spleen, liver, and lymph nodes. Those that are not ingested immediately will become more adhesive for endothelium. Thus, lactadherin appears to be involved in both hemolysis and vaso-occlusion in sickle cell anemia.
Parallel Analyses Disclose Novel Genomic Imbalances in Acute Myeloid Leukemia with Complex Karyotypes.
Authors: Rucker Frank G. , Bullinger Lars , Kestler Hans A. , Lichter Peter , Dohner Konstanze , Dohner Hartmut 
Published: April 2020

Abstract
Approximately 10 to 15 % of acute myeloid leukemia (AML) cases exhibit complex karyotypes, i.e., three or more chromosome abnormalities without presence of a specific fusion transcript. To identify novel genomic regions of interest in AML with complex karyotypes we applied comparative genomic hybridization to microarrays (matrix-CGH) allowing high-resolution genome-wide screening of genomic imbalances. We designed a microarray consisting of 2799 different BAC- or PAC-vectors. A set of 1500 of these clones covers the whole human genome with a physical distance of approximately 1.5 Mb. The remaining 1299 clones either contiguously span genomic regions known to be frequently involved in hematologic malignancies (e.g., 1p, 2p, 3q, 7q, 9p, 11q, 12q, 13q, 17p, 18q) (n=600) or contain oncogenes or tumor suppressor genes (n=699).
Using this microarray platform, 83 AML cases with complex karyotypes were analyzed. Genomic losses were found more frequently than gains; the most frequent losses were deletions of 5q (71%), 17p (53%), 7q (48%); followed by deletions of 18q (30%), 16q (28%), 3p and 12q (20% each), 12p (18%), 20q (17%), and 11q (12%). The most frequent genomic gains were trisomies of 11q (39%) and 8q (31%); followed by trisomies of 1p (22%), 21q (20%), 9p (14%), 22q (13%), 13q (12%), and 6p (10%). In part, some critical segments were delineated to genomic fragments of 0.8 to a few megabase pairs in size. In lost/gained regions parallel analysis of gene expression using microarray technology detected a gene dosage effect with significant lower/higher average gene expression levels across the genes located in the respective regions. Furthermore, 47 high-level DNA amplifications in 19 different regions were identified; amplifications occurring in at least two cases mapped to (candidate genes in the amplicon) 11q23.3-q24.1 (n=10; ETS, FLI1); 11q23.3 (n=8; MLL, DDX6); 21q22 (n=5; ERG, ETS2); 13q12 (n=3; CDX2, FLT1, FLT3, PAN3); 8q24 (n=3; C8FW, MYC); 9p24 (n=2; JAK2); 12p13 (n=2; FGF6, CCND2); and 20q11 (n=2; ID1, BCL2L1). Parallel analysis displayed overexpressed candidate genes in critical amplified region, e.g., C8FW and MYC in 8q24.
In conclusion, using high-resolution genome-wide screening tools such as matrix-CGH allows to unravel the enormous genetic diversity of AML cases with complex karyotypes, and correlation with global gene expression studies facilitates the delineation of disease-related candidate genes located in the critical regions.
SCL Oncogene Perturbs HEB/E2A Function on Thymocyte Differentiation.
Authors: Tremblay Mathieu , Herblot Sabine , Desrosiers Marianne , Todorova Margarita , Paquet Eric , Major Francois , Hoang Trang 
Published: April 2020

Abstract
The SCL and LMO oncogenes are frequently activated in childhood T cell acute leukemia (T-ALL). SCL is a transcription factor of the basic helix-loop-helix (bHLH) family that forms heterodimers with other members of the family, specifically HEB and E2A. SCL can activate or repress transcription but the mechanism through which SCL functions as an oncogene remains to be clarified. Ectopic expression of SCL and LMO in the thymus of transgenic mice causes thymocyte differentiation arrest during the preleukemic phase with aberrant differentiation at the DN3-DN4 stage, prior to the acquisition of CD4 and CD8. We therefore took several approaches to define the mechanism underlying differentiation arrest in these cells. We first analyzed global gene expression of pre-leukemic DN3 thymocytes from SCLtg/LMOtg mice against their wild type littermates. We found that in this context, these oncogenes act as global transcriptional repressor as 90% of the genes with more than two fold differences are repressed when compared to wild type controls. Furthermore, we identify the HEB/E2A pathway as being targeted by SCL/LMO and used different approaches to show that the HEB/E2A activity is repressed by these oncogenes. First, using real-time quantitative PCR, we confirmed the repression of known HEB/E2A target genes (pTa and Rag1/2). In addition, we identified new HEB/E2A target genes that were confirmed as direct target through chromatin immunoprecipitation of primary thymocytes. This array also reveals that SCL associates with HEB and/or E2A on DNA to repress their function. Furthermore, we took a genetic approach to show that SCL/LMO collaborates with HEB haploinsufficiency in inducing leukemia. Our observations therefore reveal that the repression of the HEB pathway is crucial for T cell transformation. The importance of this repression is underscored by the fact that the E2A/HEB target genes that we identify here are expressed at low levels in primary leukemic cells from T-ALL patients when compared to B-ALL or AML samples. Together, these results show that SCL/LMO repress HEB/E2A activity to block T cell differentiation, an important step for T cell leukemia.
Immunotherapy of Primary Ovarian Cancer Using Autologous Cytokine Induced Killer Cells Retargeted with Bispecific Antibodies: A Preclinical Study.
Authors: Chan John K. , Hamilton Chad A. , Cheung Michael K. , Karimi Mobin , Baker Jeanette , Gall Jonathan M. , Lum Lawrence G. , Schulz Stephan , Thorne Steve H. , Teng Nelson N. , Contag Christopher H. , Negrin Robert S. 
Published: April 2020

Abstract
Cytokine induced killer (CIK) cells are ex-vivo activated and expanded CD8+ natural killer T cells that have been shown to have anti-tumor activity. This is the first study exploring cell killing of primary ovarian carcinoma with and without bispecific antibodies (BSAbs). Primary cancer cells and autologous CIK cells were collected from women with epithelial ovarian cancer. BSAbs against CA125 (BSAbxCA125) and Her2 (BSAbxHer2) were developed using chemical heteroconjugation. On FACS analysis, the expansion and stimulation of CIK cells resulted in a significant increase of CD3+CD8+ and CD3+CD56+ T cells. With enhancement by BSAbs, the mean percent lysis in a 51Cr release assay of fresh ovarian cancer cells exposed to autologous CIK cells increased from 22% (±0.3) to 89% (±2.1) at an effector to target ratio (E:T) of 100:1. Anti-NKG2D antibodies significantly attenuated the CIK activity by 57% on primary cells. In a xenograft SCID mouse model, real-time tumor regression and progression was visualized using a non-invasive in vivo bioluminescence imaging system. Four hours after CIK cell injection, we were able to visualize CD8+NKG2D+ CIK cells infiltrating Her2-expressing cancer cells on fluorescence microscopy. Mice that underwent adoptive transfer of CIK cells redirected with BSAbxCA125 (p=0.0002) and BSAbxHer2 (p=0.0002) had a significant reduction in tumor burden and improvement in survival versus those treated with CIK cells alone (p=0.03). BSAbs significantly enhanced the cytotoxicity of CIK cells in primary ovarian cancer cells and in our in vivo mouse model. The mechanism of cytolysis appears to be mediated in part by the NKG2D receptor.
Pyrimethamine Does Not Diminish Lymphoproliferation in MRL/lpr−/− Mice and Patients with Autoimmune Lymphoproliferative Syndrome (ALPS).
Authors: Rao V. Koneti , Dowdell Kennichi C. , Dugan Faith , Dale Janet K. , Pesnicak Lesley , Bi Lilia , Penzak Scott , Avila Nilo A. , Fleisher Thomas A. , Puck Jennifer M. , Straus Stephen E. 
Published: April 2020

Abstract
ALPS is an inherited disorder of lymphocyte apoptosis leading to childhood onset of chronic lymphadenopathy, hepatosplenomegaly, autoimmunity, and an increased risk of lymphoma in a subset of patients with mutations in the intracellular domain of Fas (ALPS Type Ia). Similarly, MRL/lpr −/− mice are homozygous for Fas mutations and develop massive lymphadenopathy and splenomegaly associated with hypergammaglobulinemia, glomerulonephritis, and expansion of TCR αβ+, CD4−/CD8−double-negative (DN) T cells that are pathognomonic of ALPS. There remain no proven therapies for the lymphoproliferation underlying ALPS itself, although investigators in Europe reported that some children with ALPS showed reductions in spleen and lymph node sizes while on a weekly regimen of pyrimethamine/sulfadoxine (Fansidar@) (Br J Haematol.2002; 117:176). However, in vitro studies using stimulated human PBMCs, revealed that lymphocyte apoptosis is induced by the antifolate agent, pyrimethamine (Pyr) at an EC50 of 2.5ug/mL, and not sulfadoxine. This suggested that Pyr alone might prove clinically beneficial, thus avoiding risks of allergy and hypersensitivity associated with sulfa drugs. Hence, studies were conducted in the MRL/lpr−/− murine model of ALPS with Pyr alone given 2mg/kg twice weekly by gavage for 8 weeks, and in combination with sulfadoxine. Neither Pyr nor Fansidar yielded significant shrinkage of lymphoid mass in mice. A pilot study of the safety and efficacy of Pyr was conducted under an FDA IND in 6 children with ALPS Type 1a and one with ALPS Type III (with no identifiable mutation). Aged 6 to 17 yrs (median; 13 y), these 6 males and 1 female patient were enrolled following informed consent to be treated with oral Pyr at escalating doses (25–50mg) given twice weekly for 12 weeks. Computerized tomography was done before and after treatment to determine whether these patients would show 40 and 50% reductions in the sizes of their lymphadenopathy and/or splenomegaly, respectively. Lymph node sizes were assessed as sums of the bi-dimensional products of 3 to 5 nodes. Spleen sizes were measured as products of their craniocaudal and anterioposterior dimensions at the superior mesenteric artery level, as well as spleen volumes computed by digital image processing. The effects of treatment on other well-defined laboratory features of ALPS were also assessed. Pyr was discontinued after 11 days in one patient due to skin rash. Dose escalation was interrupted transiently in 3 others because of neutropenia, aphthous ulcer, or leucopenia. There were no significant changes in study outcome measures. The aggregate experience with Pyr and Fansidar in mice, as well as with Pyr alone in ALPS patients (Table) lead us to conclude that these agents do not manifest sufficient effectiveness to warrant larger trials.
The Novel, Orally Active Proteasome Inhibitor, NPI-0052, Induces Apoptosis in Leukemia Lymphoma Cell Lines and Patient Specimens.
Authors: Miller Claudia P. , Ban Kechen , Ruiz Stacey L. , Neuteboom Saskia , Palladino Michael , McConkey David , Chandra Joya 
Published: April 2020

Abstract
NPI-0052 is the second proteasome inhibitor with potential for clinical use since the FDA approval of bortezomib. NPI-0052 is a novel, orally active, non-peptide small molecule inhibitor discovered by Nereus Pharmaceuticals during the fermentation of a new marine Gram-positive actinomycete, Salinospora sp. In human erythrocyte derived 20S proteasomes, with EC50 values in the picomolar and nanomolar range, NPI-0052 inhibits all three proteolytic activities: the chymotrypsin-like, the trypsin-like and caspase-like activities. In the present study, exposure of 1uM NPI-0052 for 1 h inhibited the chymotryptic and caspase-like activities by greater than 90% in Jurkat and ML-1 cells. The trypsin-like activity was also inhibited to a lesser extent. NPI-0052 demonstrated varying degrees of apoptosis in cell lines representative of AML (ML-1), ALL (Jurkat), and Burkitt lymphoma (BL-41) and in mononuclear cells isolated from CLL and ALL patients, as measured by propidium iodide staining and subsequent FACS analysis. In addition, treatment of Jurkat cells with NPI-0052 resulted in activation of caspase-3 and cleavage of poly ADP-ribose polymerase (PARP). Further experiments revealed that caspase activity might be initiated differently in myeloid versus lymphoid leukemia cell lines. NPI-0052 caused cleavage of caspase-8 as demonstrated by SDS-PAGE analysis and when combined with an inhibitor specific for caspase-8 (IETD-fmk), Jurkat cells (of lymphoid origin) were protected against NPI-0052 induced apoptosis whereas ML-1 (of myeloid origin) were not. The cleaved product of Bid was detected by immunoblotting in NPI-0052 treated Jurkat cells, suggesting amplification of caspase-8 activity through mitochondria. NPI-0052 induced loss of mitochondrial membrane potential and release of cytochrome c in Jurkat cells. Cell lines of lymphocytic origin exposed to 4h of NPI-0052 resulted in increased levels of peroxide and superoxide prior to cell death. Furthermore, the antioxidant, N-acetyl cysteine (NAC), conferred protection in Jurkat cells against NPI-0052 induced apoptosis. CLL and Ph+ ALL patient material confirmed that lymphocytes from these patients are protected from NPI-0052 induced apoptosis by antioxidants. In summary, NPI-0052 inhibits all three major proteolytic activities of 20S proteasome in leukemia cells and induces apoptosis in leukemic cells and patient samples. The cytotoxic effects of NPI-0052 in leukemia and lymphoma cells warrant further testing to determine if this compound is clinically effective.
Inhibition of AKT/mTOR Signaling Induces Cell Cycle Arrest and Apoptosis in Mantle Cell Lymphoma.
Authors: Peponi Evangelia , Drakos Elias , Reyes Guadalupe , Leventaki Vassiliki , Medeiros L. Jeffrey , Rassidakis George Z. 
Published: April 2020

Abstract
Mantle cell lymphoma (MCL) is a distinct type of B-cell lymphoma associated with aggressive clinical course. The PI3K/AKT pathway is activated in a variety of human malignancies including hematopoeitic tumors. Activated (serine 473-phosphorylated) AKT (pAKT) is believed to mediate its effects, at least in part, via phosphorylation/activation of the mammalian target of rapamycin (mTOR). However, the biologic significance of AKT/mTOR signaling in MCL is unknown. We hypothesized that the PI3K/AKT signaling pathway is activated in MCL and contributes to tumor cell survival through activation of mTOR and its downstream effectors 4EBP1, p70S6K, ribosomal protein (rp) S6 (rpS6) and eIF-4E. We used 3 mantle cell lymphoma cell lines (Mino, Jeko and Z-138), a PI3K inhibitor (LY294002), and small interfering RNA (siRNA) specific for mTOR gene to investigate the biologic effects after inhibition of AKT/mTOR signaling in MCL. Treatment of MCL cells with LY294002 resulted in apoptotic cell death in a dose-dependent manner that was associated with downregultion of the anti-apoptotic proteins cFLIP and Mcl-1. Inhibition of PI3K also resulted in decreased cyclin D1 levels and cell cycle arrest as shown by decreased S-phase fraction assessed by BrdU incorporation. Changes in cell cycle and apoptosis were associated with decreased phosphorylation (activation) of mTOR and its downstream targets 4EBP1, p70S6K, and rpS6. Silencing of mTOR expression, using mTOR-specific siRNA also decreased phosphorylation of mTOR signaling proteins and induced cell cycle arrest and apoptosis in MCL cell lines. These biologic effects were associated with downregulation of cyclin D1 and modulation of apoptosis regulators. In addition, expression of activated AKT, and effectors downstream of AKT/mTOR pathway (p-p70S6K, p-rpS6, 4EBP1, eIF-4E) was assessed in MCL tumors. Using a 10% cutoff to define positivity, 10/20 (33%) expressed Ser473pAKT, 22/22 (100%) p-p70S6K, 5/20 (25%)p- rpS6, 13/14 (93%) 4E-BP1, and 16/29 (55%) eIF-4E. p-p70S6K phosphoprotein was detected predominantly in the nucleus of tumor cells. All other phosphoproteins were expressed with a cytoplasmic pattern. We conclude that activation of PI3K/AKT may contribute to cell cycle progression and tumor cell survival in MCL cells through activation of the mTOR signaling pathway. Our data suggest that inhibition of mTOR with rapamycin analogues currently used in clinical trials may be a novel therapeutic strategy for patients with MCL.