bluuebunny/crossref_metadata_2025_split huggingface dataset:
Defective Peripheral Blood Endothelial Cell Progenitors in Patients with Scleroderma and Psoriatic Arthritis
Authors: Kourelis Taxiarchis V , Manola Akrivi D , Adamidou Despoina , Sakkas Lazaros , Mperou Eyagelia , Karras George E , Vassilopoulos George , Stathakis Nikolaos E , Alexandrakis Michael G , Kyriakou Despoina S 
Published: April 2020

Abstract
Vasculogenesis is known to be defective in patients with scleroderma (SS) and psoriatic arthritis (PA) with vessel loss in the former and hypertrophic blood vessels in the latter in affected areas. We studied the number and function of peripheral blood endothelial progenitors (PBEP) in patients with SS and PA to elucidate the mechanism of EC dysfunction.
Materials and Methods: Eleven patients with SS, 13 patients with PA and 7 healthy individuals were studied. We measured CD133+/CD146+ cells in peripheral blood (PB) by immunofluorescence. We performed cell cultures of isolated CD34+ cells in endocult medium and examined the endothelial colonies (EPC). We also performed cocultures of CD34+ and autologous bone marrow stromal cells (BMSC) in double chambers. We also performed cocultures of BMSC from patients and normal endothelial cells from cord blood.
Results: The number of CD133+/CD146+ cell in PB was increased the 2 groups of patients compared to controls. The number of EC colonies in endocult did not differ in the 3 groups. The presence of autologous serum within the culture medium reduced the number of colonies in 3 patients with SS. The number and the size of EC colonies from SS patients in vitro were significantly reduced (p&lt;0.01) after co-cultures of autologous BMSC with CD34+ in culture plates with insert while they were increased (p&lt;0.01) from patients with PA. The same was true when cord blood CD34+ cells were cultured in endocult medium in the presence of BMSC of SS and PA patients.
Conclusion: EC progenitors from patients with SS and PA are increased in PB and develop normal EC colonies in vitro. They developed decreased colonies in SS and increased colonies in PA when cultured together with with autologous BMSC. This means that possible cell-cell or humoral interactions between EC and some cellular component within BMSC affect the survival and differentiation of EC.
HLA Mismatched MSC Suppress T Lymphocyte Allo responses in Vitro and Do Not Induce Immunological Memory in Recipients of MSC Infusion
Authors: Sundin Mikael , Barrett A. John , Ringdén Olle , Uzunel Mehmet , Lönnies Helena , Dackland Åsa-Lena , Christensson Birger , Le Blanc Katarina 
Published: April 2020

Abstract
Multipotent mesenchymal stromal cells (MSC) are increasingly used to treat refractory graft-versus-host-disease and other complications in HLA-matched and mismatched hematopoietic stem cell transplantation (HSCT) patients. We evaluated immunogenicity of human allogeneic MSC infused post-transplant to HLA-mismatched, i.e. patients undergoing HSCT. We compared recipient lympho cyte response to MSC and peripheral blood lymphocytes (PBL) from the MSC or third party donors before and after infusion, and lymphocyte responses to MSC and to PBL from the MSC donor in primary and secondary challenge using 3H-tymidine. MSC recipients displayed in median responses less than 500 counts per minute (CPM) to infused third party MSC 1 week to 6 months following infusion. However, the recipients responded normally to MSC donor lymphocytes, &gt;1000 CPM (p&lt;0.005), and third party lymphocytes, &gt;5000 CPM (p&lt;0.005). MSC failed to prime responder lymphocytes to rechallenge with PBL, as the response was &lt;10 000 versus &gt;30 000 CPM for the corresponding control (p&lt;0.05). On MSC rechallenge of lymphocytes primed with PBL from the MSC donor, only responses &lt;500 CPM occurred. MSC upregulated lymphocyte gene expression of CD25, IFN-γ, FoxP3, CTLA-4 and IL-10 upon MSC/PBL co-culture and MSC presence in mixed lymphocyte cultures. Unprimed and primed responder lymphocytes expanded and proliferated poorly to MSC as stimuli, evaluated by flow cytometry. The MSC failed to induce CD25+ (activated) or CD57+ (effector) CD4+ or CD8+ subsets and only inconsistently induced FoxP3+ regulatory T lymphocytes. These results confirm in vivo and in vitro that infused MSC are weakly immunogenic and do not induce significant immunological memory in HLA-disparate recipients.
Mesenchymal Stem Cells (MSC) Promote Aggressive Behavior of Human Breast Cancer Cells (MCF-7) in Vitro- the Role Cytokines (TNF-alpha) and Chemokines
Authors: Cerny Jan , Liu Yingwang , Moore Nathan , Bowen Glennice , Cerny Anna M , Kurt-Jones Evelyn , Erdman Susan , Houghton JeanMarie 
Published: April 2020

Abstract
The tumor microenvironment contributes to local tumor growth and may set up a metastatic niche for breast cancer cells. We tested the effects of spontaneously transformed murine mesenchymal stem cells (stMSC; Li et al. Cancer Res 2008) on the migration, proliferation and cell cycle of the MCF-7 human breast cancer cell line.
MCF-7 cells were collected from plates held at confluence to establish cell cycle arrest and then grown either in control media (50/50 mix of fresh and conditioned MCF-7 media) or modification of stMSC conditioned media (50/50 mix of fresh and stMSC conditioned media with TNF-alpha, anti-TNF-alpha or isotype control). MCF-7 cells were plated at equal concentrations into 24-well dishes to establish near confluent plates. The MCF-7 monolayer was wounded using a pipette tip, and the area evaluated/photographed at 0, 24 and 48 hours (in triplicate). Media was changed daily. Synchronized cultures were grown in control or MSC media for 0, 24 or 48 hours in triplicates and analyzed by FACS for cell cycle status and standard cell counts. MCF-7 cells grown in MSC cultured media healed 49% (based on wound size) while control wound size was 26% healed at 24 hrs (p=0.00591). At 48 hrs, MCF-7 cells exposed to stMSC media showed 95% healing while MCF-7 cells in control media still retained 50% of the original wound size (50% healing; p=0.00002). The exposure MCF-7 grown in control media and MSC media did not differ in cell cycle progression. MSC media increased proliferation of MCF-7 cells at 24, 48 and 72 hours compared to control media. (p&lt;0.002). In a separate experiments media from stMSC exposed to TNF-alpha led to increased healing of MCF-7 monolayer compared to stMSC conditioned media at 24 hrs (59.5% vs 44.5%; p=0.027). Healing was lower in media from stMSC cultured with anti-TNF blocking antibody compared to stMSC conditioned media at 24 hrs (66.6% vs 95.7%; p=0.002). We have also detected that stMSC conditioned media contains very high levels of TNF-alpha, MCP-1, CCL-5 and also high levels of NF-kB activity in stMSC conditioned media.
StMSCs produce factors (such as TNF-alpha) that provide breast cancer cells (MCF-7) with migration and growth advantage. Animal experiments assessing the ability stMSCs to contribute to invasion and migration of breast cancer cells are currently underway.
C/EBPalpha Is a Key Transcriptional Regulator of the Triggering Receptor on Myeloid Cells-2 Gene
Authors: Mancarelli Maria Michela , Frusciante Lily Catherine , Cauvi Gabrielle Francoise , De Rossi Charles , Torbett Bruce Edward 
Published: April 2020

Abstract
Triggering receptor on myeloid cells-2 (TREM-2) is a member of the innate immune signaling receptor TREM family. After differentiation of monocytes, TREM-2 is expressed on the cell surface of macrophages, monocyte-derived dendritic cells, microglia, and osteoclasts. TREM-2 is necessary for modulation of cellular activation and function, and loss of TREM-2 results in the decrease of phagocytosis of apoptotic neurons and cells and an increase in inflammation response of macrophage-lineage cells. To better understand TREM-2 gene regulation during monocyte differentiation we isolated and then analyzed the putative human TREM-2 promoter to identify transcriptional control. The 1131 bp TREM- 2 promoter contains binding sites for C/EBPα and PU.1 transcription factors, known for regulation of myeloid cell differentiation and function. A series of 5 promoter reporter deletion mutants were generated to dissect transcription regulation. We progressively deleted the full-length promoter, −1002/+129, of HIF1, STAT5, GATA1, YY1F, C/EBPα binding sites generating −866/+129, −632/+129, −526/+129, −258/+129, +37/+129 promoter reporters, respectively. All TREM-2 promoter reporters showed minimal transcriptional activity when transfected in 293T-HEK cells. In contrast, C/EBPα was necessary and sufficient for TREM-2 promoter reporter activity in co-transfection studies, demonstrating a 5-fold induction of transcriptional activity as compared to controls. Consistent with the TREM-2 reporter findings showing C/EBPα mediated activity, EMSA assays demonstrated specific C/EBPα binding to sites within the TREM-2 promoter. To determine if both C/EBPα binding sites in TREM-2 were necessary for activity we generated loss of C/EBPα binding site reporters and observed that only the −43/−28 C/EBPα binding site was essential for activity. To clarify if +73/+93 PU.1 binding site had a role in TREM-2 regulation, we co-transfected the full-length promoter with PU.1 and C/EBPα and observed a 10-fold increase of induction of TREM-2 transcriptional activity. However, PU.1 alone did not activate the full-length TREM-2 promoter or bind to the PU.1 consensus site, and loss of the PU.1 binding site did not alter transcriptional activity. These results suggest a key role for direct C/EBPα and indirect PU.1 participation in regulation of TREM-2. Studies are underway to precisely understand TREM-2 gene regulation and how TREM-2 controls macrophage-mediated phagocytosis and inflammation.
Characterization of Tissue-Specific HIF-1 and HIF-2 Regulatory Elements of the Erythropoietin Gene
Authors: Yoon Donghoon , Kim Hyojin , Jang Minyoung , Song Jihyun , Arnold Gregory E , Elliott Steve , Prchal Josef T. 
Published: April 2020

Abstract
Hypoxia regulates erythropoiesis and other essential processes via hypoxia-inducible transcription factors (HIFs). HIFs are heterodimers that consist of an α subunit (3 isotypes with significant homology; HIF-1α, HIF-2α, HIF-3α), and a common b-subunit; HIF-1 and HIF-2, in some instances exhibiting tissue- and gene-specific gene regulation. Erythropoietin (EPO) was the first identified HIF-1 target gene with the defined HIF-1 binding sequence. However, subsequent works suggested that HIF-2 also regulates EPO transcription and that there are other regulatory elements of EPO gene (i.e. Kidney Inducible Element KIE, Negative Regulatory Element NRE, and Negative Regulatory Liver specific Element NRLE). In silico analysis of the human EPO genome found two additional potential HIF-binding elements in the KIE and NRE regions. The comparative analysis of phylogenically conserved sequences of human, mouse, dog, and rat Epo genes further refined these mouse Epo gene HIF-binding elements as mKIE, mNRE1, mNRE2, and mNRLE2.
We treated mice in hypoxia chamber (8% O2) and monitored changes of Epo mRNA levels in liver, kidney, brain, spleen, and bone marrow. All tested tissues increased Epo transcription during hypoxia. Bone marrow, spleen, kidney, and brain showed a peak of induction of Epo transcript at 3 hours of hypoxia treatment, while liver reached the highest level at 6 hours. Mice were sacrificed and organs were harvested, and in vivo chromatin immunoprecipitation (ChIPs) was performed with antibodies against HIF-1α and HIF- 2α and tissue-specific binding regions were defined. The results from these studies are summarized below.
HIF-1 mKIE rnNRE mNRE2 mNRLE2 Norm Hyp Norm Hyp Norm Hyp Norm Hyp Liver − + − − + − ? ? Kidney − + − − + − + − Brain − + − − − + − + BM − + − − − − − + Splsen − + − − − − − + HIF-2 mKIE mNRE mNRE2 mNRLE2 Norm Hyp Norm Hyp Norm Hyp Norm Hyp “+” denotes presence and “-” absence of binding of HIF-1 and HIF-2, “?” – indicates inconclusive results. “Norm” - normoxia, “Hyp” - hypoxia. Liver − + − − − + − + Kidney + − − − + − ? ? Brain − − − − − − − + BM − − − − − − + − Spleen − + − − − − − +
In conclusion, we demonstrate the differential hypoxia-induced binding of HIF-1 and HIF-2 at different HIF binding elements in the tissues known to express Epo. Further studies will be required to define the function of these HIF-1 and HIF-2 binding elements in tissue specific Epo expression and their role in health and disease.
The Prognostic Factor Evaluation of Febrile Neutropenia Before Chemotherapy to Hematologic Malignancies
Authors: Kim Byung Soo , Choi Chul Won , Kim Seok Jin 
Published: April 2020

Abstract
Purpose Febrile neutropenia is one of the major toxicities-associated with chemotherapy especially in hematologic disorders. Thus, the occurrence of febrile neutropenia significantly can affect their treatment outcomes. But, it is difficult to predict their clinical courses and treatment outcomes. In this study, we analyzed the prechemothepeutic clinical characteristics of febrile neutropenia and their relationship with the prognosis of febrile neutropenia in the patients with hematologic malignancies.
Methods We retrospectively analyzed 259 cases of febrile neutropenia developed after the chemotherapy for their hematologic malignacies from January 2006 to July 2008 at the Korea University Medical Center to identify the potential prognosis predicting factors. For the early detection of the high risk of febrile neutropenia, we focused on the findings before chemotherapy.
Results With univariate analysis, age, underlying disease, recovery of neutropenia, onset time of febrile neutropenia, onset location, the duration of fever, infection sites, and type of cultured organism were significantly associated with mortality (p &lt; 0.05). Also, the risk of mortality was significantly associated with laboratory findings (hemoglobin, platelet count, BUN, AST, albumin, sodium, bicarbonate, PT, ESR, CRP) of prechemotherapy. In multivariate analysis, only three variables of laboratory findings were associated with poor outcomes: albumin, bicarbonate and C-reactive protein of prechemotherapy. The recoveries of neutropenia and respiratory tract infection were also the significant risk factors in multivariate analysis. The complication rate in the patients who had three poor variables (low albumin & bicarbonate level and high CRP level) was 82.8%, while in the patient who had no poor variables was only 5.7%.
Conclusion The levels of albumin, bicarbonate and CRP, in prechemotherapic condition were associated with the prognosis of febrile neutropenia in our study. Therefore, the further prospective studies will be warranted to confirm the result of this study.
High Expression of IL-15 Correlates with Disease Recurrence of Adult B-Lineage Acute Lymphoblastic Leukemia
Authors: Wu Shuling , Fischer Lars , Burmeister Thomas , Schwartz Stefan , Notter Michael , Gökbuget Nicola , Hoelzer Dieter , Hofmann Wolf-Karsten , Thiel Eckhard 
Published: April 2020

Abstract
Cytokines are involved in the response to therapy and resistance of leukemic cells as well as their biological behaviour. Interleukin (IL)-15 has been shown to enhance the proliferation of both normal and malignant lymphocytes, thus suggesting a potential role in leukemogenesis and disease progression. In this study, we analyzed the expression of IL-15 in leukemic cells from 87 patients with adult acute lymphoblastic leukemia (ALL) at initial diagnosis. Expression of IL-15 was measured by real-time RT-PCR and normalized to the internal standard b-actin. mRNA levels were correlated with protein expression by western blot analysis. We found that expression of IL-15 correlated significantly with immunophenotype: T-lineage ALL (n=23) had a more than 4-fold higher IL-15 mRNA expression as compared to B-cell precursor (BCP)-ALL (n=64, P&lt;0.001). IL- 15 expression was higher in common and pre-B as compared to pro-B-ALL, and CD1+ cortical T-ALL expressed IL-15 much higher than pre-T or mature T-ALL (median values: 0.152, 0.025, and 0.026, respectively). Patients with BCR-ABL+ BCP-ALL (n=18) had lower IL-15 expression compared to BCR-ABL− BCP-ALL (n=38, P=0.041). Furthermore, lower expression values of IL-15 were significantly associated with central nervous system involvement (P&lt;0.001). In contrast, higher expression of IL-15 was significantly correlated with mediastinal (P=0.001) and lymph node infiltration (P=0.051), but not with hepatomegaly and splenomegaly. Notably, high IL-15 expression in BCP-ALL correlated strongly with inferior probability of relapse free survival (pRFS) at five years: pRFS 0.17 ± 0.13 (IL-15 expression &gt; median, n=27) versus 0.47 ± 0.13 (&lt; median, n=37) (P=0.008). However, only a weak correlation of the probability of overall survival (pOS) and IL-15 expression was observed (pOS: 0.41 ± 0.11 versus 0 ± 0, P=0.188).
In conclusion, differential expression of IL-15 in adult ALL at diagnosis was associated with clinical features and outcome, in particular relapse free survival. It remains to be evaluated whether IL-15 might be a relevant therapy target, or might be used for risk stratification.
The Novel Human Ph+ Acute Lymphoblastic Leukemia Cell Line PhALL3.1 Is Amenable to Retroviral Gene Transfer and Mimics Human Disease Progression in SCID-Beige Mice
Authors: Frattini Mark G. , Santos Elmer , Heaney Mark Lawrence , Maslak Peter , Jhanwar Suresh C , Larson Steven M , De Stanchina Elisa , Clarkson Bayard , Brentjens Renier J. 
Published: April 2020

Abstract
Despite recent treatment advances, patients with Philadelphia (Ph) chromosome positive acute lymphoblastic leukemia (ALL) have a dismal prognosis. For this reason, a better understanding of the biology of Ph+ ALL, as well as novel approaches to treat this malignancy are needed. To date, there exist few primary Ph+ ALL tumor cell lines amenable to preclinical investigation of either the biology or treatment sensitivity of Ph+ ALL. In this report we describe the isolation and characterization of a novel Ph+ ALL cell line, designated PhALL3.1, obtained from a 62 year old patient who relapsed in the pleural space while receiving imatinib therapy. Tumor cells obtained from this malignant pleural effusion proliferated in standard complete RPMI media without supplementation with exogenous growth factors. The PhALL3.1 tumor expressed an unmutated p190 bcr-abl transcript, and demonstrated persistent sensitivity to both imatinib and desatinib in in vitro assays. Furthermore, we found that the PhALL3.1 cell line was readily amenable to retroviral gene transfer allowing for the modification of this line to express the green fluorescent protein-fire fly luciferase enzyme (PhALL3.1/GFP-FFL). When injected systemically into immune-compromised SCID-Beige mice, PhALL3.1/GFP-FFL tumor cells mimicked a clinical disease pattern with tumor involvement seen predominantly in the bone marrow, CNS, and spleen. This pattern of disease progression, as demonstrated by bioluminescent imaging, was further confirmed by histologic analysis. Likewise, when injected subcutaneously into immune-compromised mice, this cell line readily and reproducibly generated subcutaneous tumors. We conclude that the PhALL3.1 cell line is a novel, well characterized, cell line which readily proliferates both in vitro and in vivo, and therefore has significant value in future pre-clinical investigations into the biology of Ph+ ALL tumors as well as novel treatment approaches for this disease.
Clinical Trial of NPI-0052 (2nd generation proteasome inhibitor) in Patients Having Advanced Malignancies with Expanded RP2D Cohorts in Lymphoma and CLL
Authors: Price Timothy , Padrik Peeter , Townsend Amanda , Mainwaring Paul , Catley Lawrence , Longenecker Angie M. , Cropp Gillian , Palladino Michael A. , Lloyd G. Kenneth , Spear Matthew A. , Millward Michael 
Published: April 2020

Abstract
Background: NPI-0052 is a novel proteasome inhibitor that produces prolonged inhibition of the 20S proteasome. NPI-0052 has a novel structure leading to a unique proteasome inhibition, toxicology and effect profile. Preclinical research suggests an improved therapeutic ratio and activity in hematologic and solid tumor malignancy models. Secondary to these findings, clinical trials are being conducted in patients with myeloma, lymphomas, leukemias and solid tumors.
Materials and Methods: In this study patients with solid tumor, lymphoma or leukemia diagnoses were treated with NPI-0052 administered weekly, for 3 weeks in 4-week cycles in a 3+3 design dose escalation. The dose of NPI-0052 was escalated in 50–100% increments dependent on observed adverse events (AE). In addition to regular safety monitoring, proteasome inhibition (PI) (baseline, D1 & D15) and pharmacokinetics PK (D1 & D15) were assayed in blood. Once a Recommended Phase 2 Dose (RP2D) is identified, RP2D cohorts of 10 patients in each lymphoma and CLL are enrolled.
Results: 25 patients have been treated at doses ranging from 0.1 mg/m2 to 0.7 mg/m2. The AE profile has been very tolerable with fatigue, transient peri-infusion site discomfort and lymphopenia being commonly ascribed to NPI-0052. Whole blood pharmacokinetics were calculated for all patients on study. At the highest dose assessed PK parameters were (mean ± SD) AUCtot =215±129 ng/mL*min; Cmax =22.8 ±14 ng/mL; t1/2 =13.5 ± 9.2 mins; clearance= 7.8 ± 8.2 L/min and Vss = 132 ± 192 L. AUC and Cmax increased linearly with dose and the kinetics are apparently not dose dependant. PI has been assayed in blood, indicating a dose:response relationship with inhibition of chymotrypsin-like activity up to 100% observed and mean Day 1 inhibition of 78%. This level of proteasome inhibition is higher than that reported with standard doses of bortezomib, yet the profile of adverse drug reactions associated with bortezomib has not been observed. A total of 7 patients (33%) have had stable disease for at least 2 cycles (8 weeks; 2months), including one each with mantle cell lymphoma (4 cycles), Hodgkin’s lymphoma (4 cycles), follicular lymphoma (4 cycles) and sarcoma (5 cycles) and prostate adenocarcinoma, and two with melanoma (4 cycles).
Conclusions: NPI-0052 produces dose-dependent pharmacologic effects through the predicted efficacious range while producing a toxicity profile that is tolerable and dissimilar to that of the standard of care proteasome inhibitor bortezomib. These data have supported additional studies being initiated in hematologic malignancies and solid tumors.