bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Risk Organ Involvement on Outcome in Langerhans Cell Histiocytosis (LCH)
Authors: Katewa Satyendra , Sachdeva Anupam , Dinand Veronique , Yadav Satya Prakash
Published: April 2020

Objectives: To compare the outcome in children with low risk vs. risk LCH and to evaluate the impact of individual risk organ on outcome.

Material and Methods: Medical records of 52 patients were reviewed retrospectively (1992 to 2007). Patients were classified into 2 groups: A-low risk (bone, skin and lymph node involvement), B-risk patients (lungs, liver, spleen and bone marrow involvement). Group B further subdivided in B1-isolated lung involvement, B2-liver, bone marrow or both without lung, B3-lung involvement with liver and/or bone marrow. Patients were treated as per histiocytosis society protocols.

Results: There were 31 Males and 21 Females. Mean age at presentation was 4.8 yrs (range 1 month to 15.5 yrs). There were 21 patients in Group A and 31 in Group B with 0 (0%) and 6 (19%) deaths respectively. Overall mortality was 6/52 (11.5%). In group A, 6 patients had skin and lymph node involvement, 6 had unifocal bony involvement and 9 multifocal. Out of these 15 (bony involvement) patients 4 had otitis media and 5 diabetes insipidus. There were 9 patients in Group B1, 18 in Group B2 and 4 in Group B3 with mortality of 0 (0%), 4 (22.2%) and 2 (50%) respectively. Comparison of mortality in risk patients vs. low risk patients approached statistical significance (19.4% vs. 0%, p=0.069). Mortality difference within the 3-subgroups of risk patients was not statistically significant (p=0.097).

Conclusion: Outcome is worse if risk organs are involved. However, isolated lung involvement had trend towards better outcome.
The Value of Neutrophil CD64 in Early Diagnosis of Adult Sepsis
Authors: Xu Jingyan , Ouyang Jian
Published: April 2020

Purpose To observe the level of leukocyte surface antigen CD64 during adult sepsis, and to assess the value of the CD64 in early diagnosis of adult sepsis.

Methods We measured peripheral blood samples of neutrophil CD64 expression in 58 patients with suspected sepsis and 10 healthy controls.

Results The expression level of CD64 in sepsis group was significantly higher than healthy control group ( P < 0. 001). According to the ROC curve, when the best threshold in diagnosis of adult sepsis was 1.075%,the sensitivity for indentifing adult sepsis was 98. 3 % and the specificity was 90.0%.

Conclusions The expression level of CD64 will rise during adult sepsis. CD64 can be an early effective diagnosis marker in adult sepsis.
Virtual Microscopy System for Blood Morphology Evaluation and Their Applications in Hematology
Authors: Merino Anna , Gutierrez Gabriela , Garcia Jaime
Published: April 2020

We have explored the possibilities of a new technology for scanning peripheral blood slides converting the images to a digital slide that can be observed in a manner similar to a microscope.

Objectives: To evaluate the quality of peripheral blood (PB), bone marrow (BM) and body fluids (BF) images obtained in a Digital Slide Scanner in different hematological or non hematological diseases and to analyse the useful of this System in Hematology.

Material and Methods: We obtained PB samples from 13 patients with the following diagnoses: 3 acute myeloid leukemia (AML), 1 acute lymphoid leukemia (ALL), 2 B-cell neoplasm, 2 myeloproliferative disease (MPD): 1 chronic myeloid leukemia (CML) and 1 myelofibrosis, 3 Plasmodium infections (2 falciparum and 1 vivax), 1 sickle cell anemia and 1 congenital sideroblastic anemia (CSA). We also analysed 2 BM (leishmania and parvovirus infections) and 1 BF (melanoma) samples. We used a NanoZoomer Digital Pathology or NDP (Hamamatsu System, Japan) for scanning the different glass slides using 40X objective lens. A small fragment of the most appropriate area was selected and the focus was set up at different levels. Each one of the digital slides was saved in digital data and the images were analysed using the NDP software.

Results: Most values of the file sizes were approximately 300 MB for PB and BM and 50 MB for body fluid. All digital slides showed a very high image quality. It was possible to evaluate the blood cell morphology using 20X, 40X, 63X and 100X magnifications. We selected 63X as the best one for the combination size of the cells and precision in the focus. Similar to microscopic observation, myeloid and lymphoid blast cells in digital slides showed different morphologic features (cytoplasmic inclusions, primary granules, prominent vacuolation…) allowing hematological diagnosis. Myeloproliferative disorders showed a high degree of morphological abnormalities in the erythroid, granulocytic and megakaryocytic lineages. B chronic haematological neoplasms were identified in the virtual slides by morphological criteria. With respect to Plasmodium infections, the virtual images allowed examination of morphological features of both the parasites and the host red cells, facilitating the species diagnosis. Sickle cells were easily identified. Pappenheimer bodies were visible in the erythrocytes of the CSA. With respect to BM and BF, digital slides showed the morphological abnormalities for the diagnosis. The software let to calculate the diameter and the area of the blood cells.

Conclusions: Our results showed that the virtual microscopy for blood morphology evaluation can be used to substitute the glass smear samples in Hematology External Quality Assessment. In addition, the application of the NDP software may be useful in morphometric analysis of pathological cells in different hematological diseases.
Risk Factors for Febrile Neutropenia among Breast, Lung, and Colorectal Cancer Patients: A Retrospective Analysis of Health Plan Databases
Authors: Ramsey Scott Davi d , McCune Jeannine S , Blough David K , Clarke Lauren C , McDermott Cara L , Sullivan Sean D
Published: April 2020

Chemotherapy regimens, patient factors, and the use of colony stimulating factor (CSF) influence cancer patients’ risk for febrile neutropenia (FN) when they receive myelosuppressive chemotherapy. The incidence of FN and patient factors influencing that risk are relatively unknown in community settings. Using claims from Medicare, Medicaid and two private health insurance plan enrollees linked to the Puget Sound SEER registry, we examined the incidence of FN among breast, lung and colorectal cancer patients diagnosed 2002–05 who received adjuvant chemotherapy. We used logistic regression models to determine factors influencing the risk for FN within the first chemotherapy cycle, controlling for cancer stage, age, sex, race, comorbidities, chemotherapy-regimen related FN risk (as designated by the National Comprehensive Cancer Network), CSF use, health insurance type, and surgery or radiation ≤30 days from administration of first chemotherapy. Over the time horizon, 1096 breast, 1142 lung, and 755 colorectal cancer patients received chemotherapy. The incidence of any FN in the first chemotherapy cycle was (counts per 100 recipients by high, intermediate, and low-risk myelosuppressive chemotherapy according to NCCN categories respectively) 7.36, 10.0, 4.70 for breast cancer, 17.12, 14.15, 12.22 for lung cancer, and 25.0, 8.96, 6.37 for colorectal cancer. Significant predictors (p<0.05) of any FN were: breast cancer—radiation ≤ 30 days from first chemotherapy administration (OR 2.90, 95% CI 1.21–6.94), other non-black race vs. white race (OR 2.82, 95% CI 1.29–6.17), or Medicaid insurance (OR 2.31, 95% CI 1.10–4.89); lung cancer—radiation ≤ 30 days from first chemotherapy administration (OR 1.63, 95% CI 1.01–2.61), surgery ≤ 30 days from first chemotherapy administration (OR 2.08, 95% CI 1.02–4.25), Medicaid insurance (OR 2.29, 95% CI 1.08–4.84), or a Charlson comorbidity score ≥ 2 (OR 2.56, 95% CI 1.11–5.91); colorectal cancer—female gender (OR 1.86, 95% CI 1.02–3.41) or high myelosuppressive risk chemotherapy regimen (OR 7.66, 95% CI 2.95–19.89). In this analysis, predictors of FN varied between cancers. Limitations of this analysis include lack of information about chemotherapy and CSF doses, as this is not captured in the SEER registry or claims data. These results indicate that several factors may interact to influence a patient’s likelihood of developing FN in the first cycle of adjuvant chemotherapy.
The Economic Burden of Chemotherapy-Induced Toxicity in Lymphoma and NSCLC Routine Hospital Care
Authors: Ihbe-Heffinger Angela , Paessens Bernadette JL , Berger Karin , Ehlken Birgit , Shlaen Margarita , Bernard Rudolf , Peschel Christian , von Schilling Christoph
Published: April 2020

The majority of chemotherapy (CT) patients (pts) receive care outside of clinical trial settings. In contrast little is known about the economic consequences of CT-induced toxicity in routine care. This was a prospective, multi-centre, observational, longitudinal cost-of-illness study. Lymphoma and non small cell lung cancer (NSCLC) pts were enrolled consecutively from 1/2005–12/2006 at the start of first or second line (immuno) CT treatment in 4 German hospitals. Patients receiving myeloablative CT with peripheral blood stem cells were excluded. Clinical data and resource use were collected from pre-planned chart reviews. German tariffs & prices in €2007 and hospital databases were used to allocate costs to health care resources. Toxicity related costs are presented from the hospital provider perspective. 273 pts undergoing 286 treatment courses with a total of 1004 CT-cycles were evaluable. 153 pts had lymphoma (47% of courses were CHOP-like) and 120 NSCLC (78% of courses were platinum-based). Mean age was 60.1 years (SD 13.0); age≥65 years 40%; female 36%; ECOG≥2 11%; tumour stage≥3 56%; history of co-morbidity 80%. The table shows a comparison of lymphoma and NSCLC patient characteristics. 208 of treatment courses (73%) were associated with at least one hospital stay (lymphoma 69%, NSCLC 78%). Mean±SD number of inpatient days was 13.1±17.7 (lymphoma 13.5±20.7, NSCLC 12.8±14.0). Mean (median) toxicity related costs amounted to€3,624 (€1,035) per treatment course with€3,366 (€1,406) for NSCLC and€3,838 (€684) for lymphoma. 8% of CT-courses (lymphoma n=12, NSCLC n= 11) were associated with costs ≥€10,000 and accounted for 50% of total expenses. In this high cost group mean toxicity associated costs nearly doubled for lymphoma pts (lymphoma€28,607, NSCLC€15,533). Hospital basic services and personnel represented 74% of total costs (lymphoma 70%, NSCLC 80%), followed by expenses for drugs (lymphoma 15%, NSCLC 9%). Our findings highlight that toxicity management in NSCLC and lymphoma pts induces significant resource use and associated costs. Cost drivers are hospitalization and drugs. Frequency distribution of costs is asymmetric with less than 10% of CT-courses contributing to half of total economic burden. Treatment courses with mean toxicity related costs of €10,000 or more are twice as expensive for lymphoma patients as for NSCLC.

Lymphoma (n=153) NSCLC (n=120) Age: mean (SD) 58.2 (15.4) 63.0 (8.4) Age ≥ 65 58 (38%) 52 (43%) Male 99 (65%) 77 (64%) ECOG≥2 10 (7%) 20 (17%) Tumour stage ≥3 72 (47%) 102 (85%) History of co-morbidity 107 (70%) 112 (93%)
Repression of Mir-106b Family Members Does Not Alter Cell Cycle Progression in Hodgkin Lymphoma
Authors: Gibcus Johan H , Tan Lu Ping , Schakel Rikst Nynke , Harms Geert , Moeller Peter , Poppema Sibrand , Kroesen Bart-Jan , Van Den Berg Anke
Published: April 2020

MicroRNAs (miRNAs) are 19–25 nucleotide long RNA molecules derived from precursor genes that inhibit the expression of target genes by binding to their 3′ UTR region. Expression of miRNAs is often tissue specific and miRNA profiling has shown specific miRNA expression patterns in both B-cell development and lymphomagenesis.

Hodgkin lymphoma is derived from pre-apoptotic germinal center B-cells, although a general loss of B cell phenotype is noted. Using quantitative RT-PCR and miRNA microarray, we determined the miRNA profile of HL and compared this with the profile of a panel of B-cell non-Hodgkin lymphomas (NHL). The two methods showed a very good correlation for the expression levels of the individual miRNAs. Using a large panel of cell lines, we confirmed differential expression between HL and other B-cell lymphoma derived cell lines for 27 miRNAs. The HL specific miRNAs included miR-155, miR-21 and miR-106b seed family members miR-17-5p, miR-20a, miR-93, miR-106a and miR- 106b. Next, we performed target gene validation of predicted target genes for miR-17-5p, which is highly expressed in HL. Using luciferase reporter assays with stabilized anti-sense miR17-5p oligonucleotides, we showed that GPR137B, RAB12 and RBJ are likely miR-17-5p target genes in two different HL cell lines.

Previous publications indicated that miR-106b seed family members negatively regulate the cyclin-dependent kinase inhibitor 1A (p21/CIP1) resulting in cell cycle arrest at G1. Consistent with these findings, we show that the miR-106b family members are highly expressed in L428, whereas p21 is not. However, inhibition of the miR-106b seed family members in L428 does not result in elevated p21 protein expression. Furthermore, there is no cell cycle arrest, growth reduction or increase in cell death and apoptosis after inhibition of the miR-106b seed family members. Thus, we conclude that blocking of the miR-106b seed family members does not necessarily lead to indiction of p21 protein. This suggests an additional regulatory layer of p21 expression in L428 cells.
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

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<0.01) after co-cultures of autologous BMSC with CD34+ in culture plates with insert while they were increased (p<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

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, >1000 CPM (p<0.005), and third party lymphocytes, >5000 CPM (p<0.005). MSC failed to prime responder lymphocytes to rechallenge with PBL, as the response was <10 000 versus >30 000 CPM for the corresponding control (p<0.05). On MSC rechallenge of lymphocytes primed with PBL from the MSC donor, only responses <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

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<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

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

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

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 < 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

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<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<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 > median, n=27) versus 0.47 ± 0.13 (< 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

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

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.
The Comparison of Efficacy Between MVEP Chemotherapy and R- MVEP Chemotherapy in Patients of Refractory and Relapsed Non-Hodgkin Lymphoma
Authors: Qian Meihua , Wang Wensong
Published: April 2020

To observe the efficacy and toxicity of two groups of MVEP Chemotherapy and R-MVEP Chemotherapy in patients of Refractory and relapsed Non-Hodgkin Lymphoma. 19 cases patients of refractory and relapsed Non-Hodgkin Lymphoma, 9 females and 10 male, between 17–75 years of age (mean age 45.5 years), 2 cases IIa,, 11caseIII (9IIIa and 2IIIb), 6 cases IV(4 IVa and 2 IVb).Most of patients showed the enlargement of superficial and/or deep multiple lymph nodes, 4 cases had fever and the enlargement of spleen and/or liver, 2 cases with hydrothorax and ascites, 3 cases with bone marrow infiltration and 1 case with multiple lumps in liver, other 1 case with autoimmune hemolytic anemia (AIHA)and one with pericardial effusion. All patients were treated with chemotherapy included COP (or COPP), CHOP, BCAOP (or BECAOP), etc. and 2 patients of them Had been treated by radiotherapy before they were treated by MVEP Chemotherapy and R- MVEP Chemotherapy. In this experiment, A group of 13 patients were treated by MVEP Chemotherapy and G-CSF ( MVEP, Mitoxantrone 8mg/m2/d1, VP16 O.1/d1 ‘d5 (or,Wumon 0.1/d1 ‘d3),Vinorelbinum 20mg/m2/d1,Prednison 60mg/d1 ‘d7,or DXM 8mg/m2d1 ‘d7), other group of 6 patients were therapied by Rituximab plus MEVP Chemotherapy and G-CSF,( R-MVEP, Rituximab 375 mg/m2/d1,Mitoxantrone 8mg/m2/d3, Wumon 0.1/d3 ‘d5 (or VP16 O.1/d3 ‘d7),Vinorelbinum 20mg/m2/d3,Prednison 60mg/d3 ‘d9 (or DXM 8mg/m2d3 ‘d9 ), G-CSF 100 ‘300/d were injected when the number of WBC was <2.0×109/L after chemotherapy. Each cycle was 28–35 days and assessment was given after 2 cycles. The result is, The overall response rate and complete remission was 61.5 % and 38.4% respectively in treat group of MVEP, The overall response rate and complete remission was 83.3% and 50 % respectively in group of R- MVEP. One of patients of NHL with AIHA after a treating cycle achieved complete remission(all of enlarged lymph nodes disappeared and Hemoglobin return to normal).The main toxicity was mylosuppression, the minimum of WBC after chemotherapy reached 0.5 ‘1.4*109/L, few patients vomited during chemotherapy, All toxicity was no different in two groups of MVEP and R-MVEP BOur study result indicates: MEVP Chemotherapy and G-CSF is an Effective, safety and economy on patients of Refractory and relapsed Non-Hodgkin Lymphoma, but Rituximab in Combination with MEVP Chemotherapy and G-CSF is could improve the efficacy of MEVP Chemotherapy only in patients of Refractory and relapsed Non-Hodgkin Lymphoma. The efficacy of Rituximab in Combination with MEVP Chemotherapy is much better than that of MVEP Chemotherapy only on patients of Refractory and relapsed Non-Hodgkin Lymphoma, the treatment method of R-MVEP are suitable for old patients.
Characterization of Antibodies Direct against the Idiotypic VK3 Chain of HCV-Related Type-II Mixed Cryoglobulinemia and B-Cell Proliferations
Authors: De Re Valli , Simula Maria Paola , Pavan Alessandro , Marin Dolores , Caggiari Laura , Toffoli Giuseppe
Published: April 2020

Autoimmune type-II cryoglobulinemia (MC) is sustained by clonal/oligoclonal B-cell populations such the disease may be considered an “indolent B-cell lymphoma (NHL) “ and may favor overt NHL development. HCV-antigen driven mechanism induces B-cell proliferations. Clonal B-cells demonstrate a restricted used of variable genes to construct the B-cell receptor (BCR) and a homology between BCR functional regions and autoimmune rheumatoid factor (RF) activity. We underline the BCR unique repertoire with frequent rheumatoid factor activity also in other autoimmune disorders associated to NHL development as Sjogren’s syndrome and rheumatoid arthritis. All together these BCRs are characterized by their high degree of idiotypic (Id) cross reactivity. Particularly the K chain V3-20/15 is frequently found in subject with positive HCV antibody. Id is a clonamarker expressedby B cells, thus is an ideal target for immunotherapy. The evidence of few Id presented in the NHL subgroup above reported constitute the rational for the development of antibodies and recombinant proteins that use shared Id among different NHL. Five monoclonal antibodies have been produced in our laboratory toward the VK3-20 region of a subject HCV+ with NHL. Epitopes recognized has been performed using epitope excision approach. A fine determination of the antibodies activities toward specific amino acids, possibly common to different individuals, are in progress. Monoclonal antibodies reactivity has been tested in vitro in ELISA, western blot and cytofluorimetry. Apoptosis and SyK/ERK phosphorylation pathways induced following BCR cross-linking of antiId murine IgG will be performed. All the antibodies were reactive in Elisa against the VK3-Ig light chain. Two of these antibodies showed a reactivity against the light chain of cryoprecipitated immunocomplexes from several patients in western blot, and by cytofluorimter they recognised two lymphoma B-cell lines.
Rituximab Therapy for HIV-Negative Multicentric Castleman’s Disease: Seven Years’ Experience at a Single Institute
Authors: Ide Makoto , Kawachi Yasunori , Izumi Yoichiro
Published: April 2020

Background: Multicentric Castleman’s disease (MCD) is an indolent lymphoproliferative disorder of unknown etiology, and is classified into three types: hyaline-vascular type, plasma-cell type, and mixed type. Recently, we reported the use of rituximab therapy for HIV-negative patients with MCD, after obtaining the informed consent of the patient and the approval of the Institutional Review Board (Br J Haematology 121, 818–819, 2003, Eur J Haematology 76. 119–123. 2006). Therefore, we tried to review the long-term follow-up (nine to 75 months) of all patients at a single institute.

Patients and Methods: Over seven years, 5 HIV-negative patients with MCD (3 men and 2 women; median age: 39, range: 26–61 years) were referred to Takamatsu Red Cross hospital. After clinical evaluation (chest and abdominal computed tomography scans, magnetic resonance imaging and Ga67 citrate scintigrams), lymphnode biopsy was performed for histological diagnosis. Rituximab was administered intravenously at a standard dose of 375 mg/m2 weekly, for 4 or 8 times without chemotherapy, and we analyzed the clinical course of MCD after rituximab therapy. During the follow-up, clinical symptoms were recorded. At a median follow-up of 49 months, biologic parameters (complete blood count, C-reactive protein, immunoglobulin G), and cytokines (soluble interleukin-2 receptor, Interleukin-6) were determined at different time points by standard procedures.

Results: Three of the five patients had histological evidence of hyaline-vascular type MCD, and two patients had plasma-cell type MCD. Two (both with hyaline vascular type) of the five MCD patients (40%) with MCD achieved almost complete remission with rituximab therapy when followed up for 50 to 75 months. Three (one with hyaline vascular type and two with plasma-cell type) of the five patients did not show clinical remission after riruximab administration.

Conclusion;Rituximab treatment provides a durable response in a some MCD (especially those with hyaline-vascular type) patients, but is not always successful.
A Novel Clinical Syndrome Associating Severe Congenital Neutropenia and Complex Developmental Aberrations Caused by Deficiency of G6PC3
Authors: Boztug Kaan , Appaswamy Giridharan , Ashikov Angel , Schäffer Alejandro A , Salzer Ulrich , Diestelhorst Jana , Germeshausen Manuela , Brandes Gudrun , Lee-Gossler Jacqueline , Noyan Fatih , Gatzke Anna-Katherina , Minkov Milen , Greil Johann , Kratz Christian , Petropoulou Theoni , Pellier Isabelle , Bellanne-Chantelot Christine , Rezaei Nima , Mönkemöller Kirsten , Irani-Hakimeh Noha , Bakker Hans , Gerardy-Schahn Rita , Zeidler Cornelia , Grimbacher Bodo , Welte Karl , Klein Christoph
Published: April 2020

We here describe a previously unrecognized nosological entity in 12 patients from 8 unrelated pedigrees. All patients presented with severe congenital neutropenia and severe invasive bacterial infections. In addition, patients had a variety of additional syndromic features such as congenital heart disease (8/12), urogenital malformations (5/12), inner ear hearing loss (2/12), and myopathy (1/12). Furthermore, most patients (10/12) showed increased visibility/angiectasia of subcutaneous veins. The bone marrow smear was characterized by a typical “maturation arrest” due to premature apoptosis of mature neutrophils. Similar to Kostmann’s disease secondary to mutations in HAX1, myeloid cells from patients with this novel syndrome showed increased susceptibility to apoptosis. Myeloid progenitor cells revealed an abnormally enlarged rough endoplasmic reticulum and increased endoplasmic reticulum stress evidenced by increased expression of BiP. A genome-wide linkage study, performed in two consanguineous pedigrees, gave statistical evidene of a linkage interval on chromosome 17q21 (LOD score 5.74). We identified homozygous missense mutations in G6PC3, a ubiquitously expressed paralog of glucose-6-phosphatase. Biochemical studies confirmed deficient enzymatic activity. Using retroviral G6PC3-gene transfer into primary hematopoietic stem cells and in vitro differentiation into myeloid cells, the phenotype of increased susceptibility to apoptosis could be reverted. Eight distinct biallelic mutations were found, including missense and nonsense mutations. G6PC3-deficient myeloid cells showed a predominance of the unphosphorylated form of GSK3beta, a key molecule controlling cellular differentiation and apoptosis. As a consequence of increased GSK3beta activity, increased phosphorylation of the antiapoptotic molecule Mcl1 was detected, explaining increased susceptibility to apoptosis in neutrophils. In summary, our study describes a novel disease, determines its molecular etiology, and sheds light on the role of glucose-dependent pathways in controlling the homeostasis of the endoplasmic reticulum and control of apoptosis.