bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Transforming Growth Factor Receptor Type II in Patients with Agnogenic Myeloid Metaplasia (AMM).
Authors: Hemavathy Kirugaval C. , Novetsky Allan D. , Wang Jen C. 
Published: October 2019

Abstract
We previously reported that blood TIMP-1 (Tissue Inhibitor of Metalloproteinase), OPG (Osteoprotegerin), and TPO (Thrombopoietin) levels are increased and blood MMP-3, and bone marrow SDF-1 levels are decreased in patients with AMM. Enhanced TGF-ß 1 activity with Transforming Growth Factor Receptor Type II (TGF ß RII) abnormalities is likely to be associated with these findings. It has been previously reported that TGF-ß RII levels are decreased in AMM patients. The current study was undertaken to explore the defects in the TGF-ß1 pathway in this disease. We cloned and sequenced the full length TGF ß RII cDNA from AMM patients and no mutations were detected on sequencing. We then determined the expression levels of TGF ß RII in the CD34+ progenitor cells by Real-Time RT-PCR and found it to be reduced in patients with AMM compared to normal volunteer CD34+ cells used as controls. Hence we cloned the promoter region of TGF ß RII (−1233 to +50) from the genomic DNA of Peripheral Blood Monocytes by PCR, using gene specific primers. On sequencing, no mutations were detected in the promoter regions. The same promoter region when cloned upstream of a luciferase gene, drove the transcription of the reporter gene similar to the levels observed with the promoter from controls. This augmented the absence of any structural abnormalities in the region of the TGF b RII promoter. We then analyzed the methylation status of TGF ß RII promoter using the technique, Sodium Bisulfite-PCR for Sequencing. Genomic DNA was modified using Sodium Bisulfite and the TGF ß RII promoter was amplified by PCR with Primers capable of amplifying both unmethylated and methylated DNA. The modified PCR products were cloned into TOPO TA sequencing vector from Invitrogen, CA and sequenced. The sequencing results revealed absence of any CpG hyper-methylation in the promoter region of TGF ß RII. To account for reduced TGF ß RII expression in the absence of coding region abnormalities, promoter abnormalities or hypermethylation, we determined Histone Deacetylase Activity HDAC activity in the CD34+ cells of AMM patients. There was an indirect correlation between HDAC activity and TGF ß RII mRNA levels implying HDAC mediated silencing of the TGF ß RII promoter in AMM patients.
Plasma and Intracellular Concentration of TNF and Its Receptors in Peripheral Blood and Bone Marrow in Different Stages of B-CLL.
Authors: Rolinski Jacek , Bojarska-Junak Agnieszka , Hus Iwona , Dmoszynska Anna 
Published: October 2019

Abstract
TNF has been proposed to play a role in the regulation of growth and death of leukemic B-CLL cells. However, the biological effects of TNF on leukemic cells, as well as its role as a prognostic factor need to be further investigated. The aim of the study was to eevaluate the correlation of TNF and its receptors in peripheral blood (PB) and bone marrow (BM) with the stage of B-CLL and some other clinical parameters. PB and BM were taken from 44 newly diagnosed, untreated B-CLL. patients. The control group consisted of 20 healthy subjects. We used flow cytometry technique to assess the capability of T and B lymphocytes to produce TNF and ELISA method to measure plasma levels of TNF and their soluble receptors. We found, that PB and BM plasma TNF concentration in the patients was significantly higher than in the healthy control (2.61 pg/ml. vs 0.62 pg/ml; and 2.91 pg/ml vs 0.75 pg/ml, respectively p<0.001). TNF concentration in PB and BM was significantly higher in Rai stage III–IV than in early stages (p<0.01). There was a correlation between the PB and BM TNF level and lymphocytosis (p<0.005) and the total tumor mass (TTM) (p<0.0001). The PB and BM TNF concentration positively correlated with the percentage of T CD3+ lymphocytes producing intracellular TNF (p<0.01).
The percentage of T cells from PB an BM expressing cytoplasmic TNF was significantly higher in patients (PB:39.11±16.97%; BM:40.73±18.19%) than in normal controls (PB:15.74±7.95%; BM:18.80±12.93%) (p< 0.00001; p<0.005, respectively). In PB and BM from B-CLL patients the percentage of CD3+ cells expressing intracellular TNF was significantly higher than the percentage of CD19+/TNF+ cells (p<0.0001). Besides, it was found that the percentage of T cells expressing cytoplasmic TNF positively correlated with the stage of disease (p<0.01). In PB positive correlation were found between the number of T CD3+/TNF+ cells and lymphocytosis (p<0.05) and TTM (p<0.001). The percentage of leukaemic B cells positive for TNF did not correlate with the stage of disease.
There was increased expression of TNF-RI and TNF-RII in leukaemic B cells in comparison to normal B-cells was observed (p<0.0001). We found positive correlation between the number of CD5+ B lymphocytes and the levels of soluble TNF-RII (sTNF-RII) (p< 0.05). The sTNF-RII levels in PB and BM significantly correlated with the stage of disease acc. Rai (p<0.0001). Furthermore, the sTNF-RII concentration positively correlated with lymphocytosis and TTM (p<0.0001). These results strongly support the key role TNF in B-CLL pathogenesis. Our results suggest that TNF may function as growth factor for B-CLL cells. CD3+T cells may be the important source of this cytokine in advanced B-CLL. It seems that changes in T cells capability to produce cytoplasmic TNF are associated with disease progression. However, further studies are required to confirm the key role of TNF in B-CLL pathogenesis.
Fludarabine-Induced Apoptosis in Chronic Lymphocytic Leukemia Cells Is Not Affected by Inhibitor of Apoptosis Proteins (IAPS).
Authors: Silva Karina Lani , de Paula Castro Eric Delfraro , Vasconcellos Deborah Vidal , Kwee Jolie Kiemlian , Moellman Arthur Coelho , Klumb Claudete Esteves , Maia Raquel Ciuvalschi 
Published: October 2019

Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the accumulation of mature lymphocytes due to defective apoptosis. Inhibition of apoptosis also contributes to chemoresistance, mainly in patients in advanced clinical stages. Fludarabine is a potent chemotherapeutic drug used for patients with B-CLL. Despite good initial clinical responses, some patients acquire resistance to this drug. One of the mechanisms through which malignant cells are believed to acquire resistance to apoptosis is by overexpression of inhibitor of apoptosis proteins (IAPs). IAPs are a family of proteins able to block apoptosis either by binding and inhibiting caspases or through caspases-independent mechanisms. To date, overexpression of IAPs has been detected in many types of malignant diseases, including B-CLL. As fludarabine can induce apoptosis trough caspases activation and this proteases could be inhibited by IAPs, we evaluated in this current study, the expression of three members of IAP family (c-IAP1, c-IAP2 and XIAP) by immunocitochemistry in 30 peripheral blood B-CLL specimens and correlated it to fludarabine-induced apoptosis. In parallel, we also evaluated the expression of these IAPs before and after in vitro fludarabine treatment. B-CLL cells were incubated with fludarabine25 m M for 12 hours and the induction of apoptosis were measured by annexin-V assay. The apoptosis index by fludarabine was extremely significant (p < 0.001). Different levels of c-IAP1, c-IAP2 and XIAP expression were commonly found in circulating B-CLL cells. High levels of c-IAP1, c-IAP2 and XIAP expression were observed in 20%, 60,7% and 60% of samples, respectively. However, the overexpression of these IAPs was not correlated with an in vitro fludarabine apoptosis-resistance. Besides, fludarabine was not able to change the levels of IAPs expression as described for other drugs in literature. These results suggest that fludarabine might cause apoptosis by a caspase-independent manner.
Gene Expression Profiling of Plasma Cell Dyscrasias: The Role of IGH Translocations in the Heterogeneity of Multiple Myeloma.
Authors: Neri Antonino , Mattioli Michela , Agnelli Luca , Fabris Sonia , Baldini Luca , Morabito Fortunato , Bicciato Silvio , Verdelli Donata , Intini Daniela , Nobili Lucia , Cro Lilla , Pruneri Giancarlo , Callea Vincenzo , Stelitano Caterina , Lombardi Luigia 
Published: October 2019

Abstract
Multiple Myeloma (MM) is the most common form of plasma cell dyscrasia, characterized by a marked heterogeneity of genetic lesions and clinical course. It may develop from a premalignant condition (monoclonal gammopathy ofundetermined significance, MGUS) or progress from intra-medullary to extra-medullaryforms (plasma cell leukemia, PCL). To provide insights into the molecular characterization of plasma cell dyscrasias and to investigate the contribution of specific genetic lesions to the biological and clinical heterogeneity of MM, we analyzed the gene expression profiles of plasma cells isolated from 7 MGUS, 39 MM and 6 PCL patients by means of DNA microarrays. MMs resulted highly heterogeneous at transcriptional level, whereas the differential expression of genes mainly involved in DNA metabolism and proliferation distinguished MGUS from PCLs and the majority of MM cases. The clustering of MM patients was mainly driven by the presence of the most recurrent translocations involving the immunoglobulin heavy-chain locus. Distinct signatures have been found to be associated with different lesions: the overexpression of CCND2 and genes involved in cell adhesion pathways was observed in cases with deregulated MAF and MAFB, whereas genes upregulated in cases with the t(4;14) showed apoptosis related functions. In addition, we identified a set of cancer germ-line antigens specifically expressed in a sub-group of MM patients characterized by an aggressive clinical evolution, a finding that could have implications for patient classification and immunotherapy.
Higher Expression Levels of the Adhesion Molecules VLA-4 and ICAM-1 on Multiple Myeloma Cells after Chemotherapy Are Associated with Survival of a Multidrug Resistant Subpopulation in Vivo.
Authors: Schmidmaier Ralf , Mörsdorf Kerstin , Baumann Philipp , Emmerich Bertold , Meinhardt Gerold 
Published: October 2019

Abstract
Objectives: Primary drug resistance is a major problem in multiple myeloma (MM), an incurable disease of the bone marrow. Adhesion of multiple myeloma cells to bone marrow stromal cells (BMSC) has been shown to cause strong primary resistance. The adhesion molecules LFA-1 and VLA-4 are upregulated upon treatment with cytotoxic agents. Furthermore, we have shown that the corresponding ligands on HS-5 BMSCs, VCAM-1 and ICAM-1, are upregulated after incubation with melphalan, suggesting increase of adhesion mediated drug resistance after chemotherapy. In this context, the expression levels of important adhesion molecules on MM cells of consecutive MM patients before and after chemotherapy have been determined in this study.
Methods: The expression levels of VLA-1, VLA-4, VLA-5, LFA-1, VCAM, ICAM-1, CD138, CD38, and CD56 were determined on MM cell lines, HS-5 stromal cells, and primary myeloma cells of 20 consecutive patients by flow cytometry in comparison to isotype control. 9 patients had been pre-treated (mostly induction chemotherapy and high dose melphalan with stem cell rescue) and 11 patients had been at diagnosis without treatment. Interpatient comparison of treated and untreated patients was performed. Intrapatient analysis (before and after high dose chemotherapy) will be performed in the follow up.
Results: VLA-4 and ICAM-1 are upregulated after chemotherapy by 54% and 64%, respectively. Similar upregulation of CD38 could be observed (62%), whereas CD138 shows downregulation by about 50%. CD56, VCAM, and LFA-1 expression was not significantly altered after chemotherapy.
Conclusion: The adhesion molecules VLA-4 and ICAM-1, which are essential for MM-BMSC interaction, are upregulated after chemotherapy. This finding supports our preclinical data and the hypothesis, that adhered, primary drug resistant MM cells are selected by chemotherapy and herewith contribute to multidrug resistance in multiple myeloma.
Serum Soluble Syndecan-1 Levels Correlate with Erythropoietin and Inversely with Hemoglobin Levels in Untreated Anaemic Myeloma Patients: Possible Role of Syndecan-1 in the Mechanisms of Chronic Anaemia.
Authors: Kyrtsonis Marie-Christine , Vassilakopoulos Theodoros P. , Papadogiannis Argyro , Tzenou Tatiana , Dimopoulou Maria N. , Siakantaris Marina P. , Kokoris Styliani I. , Dimitriadou Evangelia , Angelopoulou Maria K. , Kalpadaki Christine , Pangalis Gerassimos A. 
Published: October 2019

Abstract
Impaired erythropoiesis with inappropriate erythropoietin (EPO) production is seen in about 40% of symptomatic multiple myeloma (MM) patients at diagnosis and worsens during terminal refractory disease. Up-regulation of proinflammatory and proapoptotic factors, namely TNF-alpha, INF-gamma, TGF-beta1, FAS-Ligand and TRAIL are involved in the pathogenesis of defective erythroid maturation. The involvement of these factors in the progression of MM has been studied and no correlation was found for TNF-alpha, INF-gamma and TGF-beta1, while it was shown that the up-regulation of FAS-L/TRAIL in MM cells contributes to MM aggressiveness. Increased soluble syndecan-1 (s-synd-1) serum levels have been shown to constitute an adverse prognostic factor for survival in MM. Furthermore at least 3 studies, including ours, also found a negative correlation of s-synd-1 levels with hemoglobin. The purpose of this study was to investigate the correlations between s-synd-1, hemoglobin (Hb), endogenous EPO levels and other parameters of disease in anaemic MM patients before any treatment including rHu-EPO administration. 41 MM patients (19 males and 22 females) with variable degree of anaemia (Hb <13.5 g/dl for men and <12.5 g/dlfor women) were studied. Median Hb was 10.5 g/dl (range 6.9–13.5); 32% had Hb below 10 g/dl. Median age was 68y. 7 patients had Durie and Salmon stage I, 21 stage II and 13 stage III. Serum EPO levels were determined by RIA and s-synd by ELISA (Diaclone Research, Besancon, France) in frozen sera of MM patients and 24 healthy individuals. Normal serum EPO levels range from 4.9 to 52 mU/ml in our laboratory. Median s-synd-1 was 36 ng/ml (9–75) in healthy individuals.The median value of EPO levels in MM patients was 40.1 mU/ml (7.2–313) and of s-synd-1 160 ng/ml (26–3500). Serum EPO levels correlated as expected with Hb (Spearman’s rho -0.223, p=0.14), creatinin (Spearman’s rho -0.32, p=0.04) but also with s-synd-1 (Spearman’s rho 0.33, p=0.03). Serum EPO did not correlate either with parameters reflecting disease mass such as beta2microglobulin (Spearman’s rho −0.19, p=0.90) and percentage of bone marrow infiltration (Spearman’s rho 0.118, p=0.48) or CRP (Spearman’s rho −0.189, p=0.29) that reflects proinflammatory cytokines’ activity. Our results in addition to the fact that s-synd-1 levels inversely correlate with Hb levels, suggest that s-synd-1 may be involved in the mechanisms leading to chronic anemia in multiple myeloma. Further studies are needed to confirm these findings.
The Clinical Characteristics and Therapeutic Status of Multiple Myeloma in China- a 23 Years Retrospective Analysis in One Single Center.
Authors: Qiu Lugui , Mai Yu-jie , Li Rei , Zou Dehui , Wang Yafei , Zhao Yao-zhong , Yang Renchi , Wang Jianxiang , Xiao Zhijian , Han Mingzhe , Qian Linsheng 
Published: October 2019

Abstract
The clinical characteristics and treatment results of the multiple myeloma patients admitted to our hospital from January 1980 to December 2002 were retrospectively analyzed and compared in order to put forward a feasible therapeutic strategy for MM in China.
Results:
1. Clinical characteristics (1) 432 cases with full follow-up statistics were analyzed. The median age was 57 years (20 to 80).The peak onset age were 50 to 60 years. The ratio of male to female was 2.35:1. (2) 81% patients had anemia(Hb<100 g/L) at diagnosis. The median proportion of tumor cells in the bone marrow was 0.38 (0.005–0.970). 72.1% with high β2-MG (>3mg/L), 60% of the patients had abnormities in X-rays. (3) M-protein peak was detected in 78.5% patients. IgG made up the majority in classification(48.1%), IgA (21.8%)and light-chain(20.5%)next to it. 144 patients were in stage IIIA(40%), and 87 cases in IIIB(24.1%)at first diagnosis. (4) The median survival duration was 27 months (1–137months).
2. Treatment results: 206 cases had integral therapeutic records in 432 patients. (1) 200 patients were administered with drugs therapy, and their 3-year and 5-year survival were 32.01% and 15.8% respectively. The OS, median survival, 3-year and 5-year survival in MP group(n=46) were 30.4%, 30 months, 21.96% and 13.2% respectively. 149 cases(72.3%)received multi-drug combined chemotherapy(CCT).The overall response was 50.3% and significant higher than that of MP group(P=0.01).While the median survival duration(30.5 months), 3-year and 5-year survival rates (35% and 16.7% respectively) were as same as MP group (P>0.05. (2). 38/200 cases (19%) were treated with interferon in combination with chemotherapy over 6 months. The overall response rates were 53.6%, and the median survival duration was 52 months, and presented an increased response(P=0.02)and a prolonged survival duration (P=0.0003) compared with the no interferon group(34.4%,27months respectively. (3)Thalidomide was administered to 29 cases(14.5%), including 5 who took it alone(2.5%). The overall response rates were 50.3%. (4) 6 patients received hematopoietic stem cell transplantation. 5 patients with autologous peripheral blood stem cell transplantation were alive averagely in 73±12.5 months by the end of the follow-up.
Conclusion:
1) MM patients in China had poor prognostic characteristics except of younger age compared with the westerns, especially most patients were diagnosed at III phase with significant lower survival.
2) CCT yielded higher response rates but no longer duration of survival benefit than MP when used as treatment for MM. INF-a and Thalidomide could benefit MM by combination with chemotherapy. Among patients who are <60 years old with good performance, HSC could prolong survival duration.
Campath-1H May Have Activity in the Treatment of Multiple Myeloma.
Authors: Gasparetto Cristina , Horwitz Mitchell E. , Gockerman Jon P. , de Castro Carlos M. , Moore Joseph O. , Smith Clay A. , Davis Patty , Lassiter Martha , Houser Lesa , Chao Nelson J. , Rizzieri David 
Published: October 2019

Abstract
Previously we have demonstrated that multiple myeloma cell lines and primary cells express CD52, the antigenic target of Campath-1H, and that they undergo apoptosis following treatment with alemtuzumab in-vitro (Gasparetto et al. ASH #3210,2002). Based on these observations, we initiated a clinical trial where patients with advanced, multiply relapsed MM were treated with subcutaneous (sc) alemtuzumab as a single agent. Nine patients (6 men/4 women, age range 45–69 yrs) were initially treated with the standard dose of 30mg sc three times per week for up to 12 weeks. All patients had at least 4 lines of prior therapy and 8/9 patients had undergone high dose chemotherapy followed by autologous peripheral blood stem cell transplant. Treatment related toxicities included pancytopenia in all patients with grade IV neutropenia in the first four patients, necessitating withdrawal from study. Subsequently, all patients were treated with G-CSF and aggressive transfusion support and no further patients were removed from study due to cytopenias. All patients were prophylaxed with Famvir, Fluconazole and Septra and no opportunistic infections were noted during treatment. Two patients developed acute renal insufficiency that reversed when alemtuzumab was discontinued. One patient completed the entire 12 weeks of treatment. One patient had a PR with a 40% reduction in M-protein after two months of treatment, which reversed once Campath-1H was discontinued. Pharmacokinetic studies of one patient demonstrated that it required 8–10 weeks of treatment using the sc protocol to achieve serum levels of alemtuzumab of 1ug/ml, the level considered to be tumoricidal in-vivo. Based on this observation, the treatment protocol was modified so that the initial week of escalating doses was given IV to achieve more rapid therapeutic levels. Following this, sc alemtuzumab was given as above. One patient with non-secretory multiple myeloma with relapsed disease following 5 prior lines of therapy has been treated with this modified protocol. Treatment with alemtuzumab and growth factor support in this patient has been well tolerated except for the development of RSV pneumonia. At 4 weeks, a PET scan demonstrated a significant overall decrease in metabolic activity in multiple bony areas consistent with a response to treatment. These initial results suggests that alemtuzumab is associated with significant toxicities in patients with advanced multiple myeloma including pancytopenia, infections and possibly renal insufficiency but all of these were reversible and have been minimized with aggressive prophylactic therapy. Alemtuzumab does appear to have modest activity against MM in this heavily pre-treated group suggesting that it should be explored in combination with other agents and at earlier stages of disease.
Human Cytomegalovirus Can Activate and Induce Apoptosis in Mesenchymal Stem Cells.
Authors: Wei Guoqing , Lin Maofang , Huang He , Cai Zhen 
Published: October 2019

Abstract
Objective Mesenchymal stem cells (MSCs) is a very important cell system in bone marrow stromal and human cytomegalovirus (CMV) infection is one of the most common complications following stem cells transplantation. Our studies were to investigate the CMV infection of MSCs as well as the apoptosis induction effects.
Methods MSCs were infected by 100,10,1 TCID50 of CMV. After 48h of culture with DMEM supplemented with 10% (v/v) fetal calf serum, MSCs shapes were observed. CMV particles and ultra structure in the cells were detected with transmission electron microscope. RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene and GAPDH gene. Flow Cytometry was used to detect the apoptotic cells.
Results After the infection of CMV, shape of MSCs changed, turning from thin shuttle shape to round and thick ball shape, even escaping from wall. Cell shape changed more dominantly as the CMV titer increased. With transmission electron microscope, CMV particles and typical cellular apoptotic character could be seen in the cells. MSCs infected by CMV could express IE mRNA compare with the uninfected and there were more apoptotic and necrosis cells in the cells infected by CMV.
Conclusion CMV can not only infect MSCs and activate in the cells but also can induce apoptosis of the cells.