bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hypoxia-Inducible Factor 1 Mediated Expression of Hypoxia-Induced Vascular Endothelial Growth Factor in Human Bone Marrow Stromal Cells, Which Is Involved in MAP Kinase and Phosphatidylinositol 3-Kinase Signaling.
Authors: Tang Guangxiao , Liu Gexiu 
Published: October 2019

Abstract
INTRODUCTION: Recent studies indicate that transplantation of bone marrow stromal cells after rat traumatic brain injury provided functional recovery. Hypoxia is the major characteristic of ischemic microenvironment. Vascular endothelial growth factor (VEGF) has been demonstrated in vivo and in vitro to be the principal mediator of hypoxia-induced angiogenesis. Recent studies indicate that VEGF implicated in neuroprotection. We tested the hypothesis that hypoxia induces expression of vascular endothelial growth factor (VEGF) in human bone marrow stromal cells (hBMSC) by hypoxia-inducible factor 1 (HIF-1).
METHODS: Human bone marrow adherent cells were cultured, and were passaged in DMEM/F12 containing 10% FBS. The fifth passage cells were identified as stromal cells. Subconfluent cells were used, and were cultured with hypoxic DMEM/F12 in hypoxia condition (94% N2 + 1% O2 +5% CO2), and were treated with 50 μM PD98059, or 200 nM wortmannin, or SiRNA specific for HIF-1mRNA. Cells were harvested after 6 h for analysis of active phosphorylated kinases (Akt and MAPK) and the HIF-1-alpha protein or after 24 h for analysis of VEGF protein and mRNA. VEGF mRNA was detected by semi-quantitative RT-PCR, and VEGF protein by ELISA, and kinases and the HIF-1-alpha protein by Western blot.
RESULTS: Hypoxia increased significantly level of both VEGF mRNA and VEGF protein in hBMSC s, which were low level in normoxic cultured cells. 24 hours after treatment, ratio of RT-PCR product between VEGF and β-actin reach to (46.25±7.54)% from (19.61±4.57)% of control cells (P<0.01). VEGF protein increased to (142.77±22.33) pg/ml from (45.85±9.69) pg/ml (P<0.01). In addition, hypoxia induced active phosphorylated kinases (Akt and MAPK), and enhanced level of the HIF-1-alpha protein. Moreover, PD98059, or wortmannin, or SiRNA specific for the HIF-1-alpha mRNA inhibited induction of the VEGF gene by hypoxia.
CONCLUSION: The expression of VEGF mRNA and VEGF increased in human bone marrow stromal cells during hypoxia condition, which is involved in MAP kinase and phosphatidylinositol 3-kinase signaling. These results support that these cells play an important role in therapy of ischemic/hypoxic injury such as cerebral ischemia and myocardial ischemia. The hMSC capacity to increase expression of VEGF may be the key to the benefit provided by transplanted hMSCs in the ischemic injuries.
A Comparison of Outcome and Second Malignancies in Adolescents and Adult Patients with Hodgkin’s Lymphoma (HL). Results from the German Hodgkin Study Group.
Authors: Bredenfeld Henning , Gilman Elena , Pfistner Beate , Diehl Volker , Engert Andreas 
Published: October 2019

Abstract
Introduction: Whether adolescent (alc) patients with HL represent a distinct patient group, requiring a therapy strategy separated from adults, is currently focussed in international debates. Most study groups are treating adult (ad) patients (pts) (age > 18 years) apart from younger patients (age < 18 years), considering a difference in treatment related toxicity and therapy outcome in both patient groups. Between 1988 and 1998, the GHSG trial generations 2 (G2) and 3 (G3) included younger patients from the age of 15 (G2), and 16 (G3), in identically therapy regimen with adults up to 65, and 75 years.
The trials for early stages HD were comparing radiation therapy (RX) arms only (HD4, recruited 1988–1993), and, RX vs 2 cycles polychemotherapy plus RX (HD7, 1993–1998). Intermediate stages HD received 2 cycles of different chemotherapy regimen plus identically RX (HD5, 1988–1993), and, 2 identically cycles of chemotherapy plus different RX (HD8, 1993–1998). Advanced stages HL were treated with either 4 cycles conventional polychemotherapy plus EF, or, 4 cycles of a hybrid regimen plus EF RX (HD6, 1988–1993); the following trial (HD9, 1993–1998) compared 3 arms of different polychemotherapy followed by RX on bulky disease.
Methods: In G2 and G3, a total of 573 adolescents (15–21 yrs) in early, intermediate, and advanced stages HL were compared with 4917 pts (15–75 yrs) for complete remission rate (CR), 5 yrs survival rate (SV), 5 yrs freedom from treatment failure (FFTF), and, secondary neoplasias (2nd NPL). (table 1)
Conclusion: Based on this large analysis of HL patients treated in prospectively randomized trials, the prognosis of patients aged 15–21 years is rather similar to the prognosis of adult patients. A more detailed evaluation including age-adapted prognosis will be presented.
Results: Adolescents /Adults CR % 5 yrs FFTF % 5 yrs SV % Alc Ad Alc Ad Alc Ad Early stages HD4 97.7 97.3 81.6 80.5 97.6 95.1 HD7 93.3 94.3 82.0 83.3 100 94.1 Intermediate stages HD5 93.5 93.4 83.3 81.7 94.3 90.5 HD8 93.8 92.6 84.5 82.7 97.3 91.3 Advanced stages HD6 77.8 77.0 61.0 58.8 85.3 78.6 HD9 93.5 90.3 85.8 79.2 92.4 88.0
Hematopoietic Cell Transplantation-Specific Comorbidity Index To Predict Non-Relapse Mortality and Survival after Allografting.
Authors: Kato Ruri , Fukuda Takahiro , Usui Eiji , Yamasaki Satoshi , Maruyama Dai , Morita Yuriko , Kim Sung-Won , Mori Shin-ichiro , Tanosaki Ryuji , Tajima Kinuko , Heike Yuji , Makimoto Atsushi , Tobinai Kensei , Takaue Yoichi 
Published: October 2019

Abstract
Pre-transplant comorbidity can affect the outcome of allogeneic hematopoietic cell transplantation (HCT). The Seattle group recently proposed a new HCT-specific comorbidity index (HCT-CI) which was based on the Charlson Comorbidity Index (Blood, prepublished online, June 30, 2005).
To validate this scoring system, we retrospectively reviewed the medical records of 315 patients with hematologic malignancies who underwent allogeneic HCT after fludarabine-based reduced-intensity (n=160) or conventional (n=155) conditioning at our center between 2000 and 2004. The median age of the patients was 46 (range, 1–68) years. The diagnoses included acute myeloid leukemia or myelodysplastic syndrome (n=153), chronic myelogeneous leukemia (n=30), acute lymphoblastic leukemia (n=36), lymphoma (n=90), and other hematologic malignancies (n=6). Donors included HLA-matched (n=120) or mismatched (n=53) relatives and unrelated volunteers (n=142). Stem cell source was G-CSF-mobilized peripheral blood stem cell (n=169), bone marrow (n=117), or cord blood (n=29). We did not include the pulmonary function test results due to our inconsistency with the test.
The HCT-CI captured 45% of patients with scores >0 (score 1, n=71; score 2, n=22; score 3, n=27; score 4, n=15; score >4, n=6). The capture rate of HCT-CI in patients who received reduced-intensity conditioning was higher than that in those who received conventional conditioning (51% vs 38%). The involved organ systems included hepatic (n=68), recent infection (n=38), prior malignancies (n=17), cardiac (n=16), renal (n=11), metabolic (n=11), psychiatric (n=11), pulmonary (n=8), and gastrointestinal (n=4) abnormalities. The Kaplan-Meier estimate of overall survival was significantly different among risk groups stratified according to HCT-CI (Figure 1, p<0.0001). In Cox proportional hazard models, a higher HCT-CI score, disease risk, and transplant from donors other than HLA-matched relatives were associated with poor overall survival. A higher HCT-CI score, greater patient age, and transplant from donors other than HLA-matched relatives were associated with a significantly increased risk for non-relapse mortality.
In conclusion, the new HCT-CI using pre-transplant variables was the most significant predictor of non-relapse mortality and survival after allografting. Our validation study suggests that this index will be a useful tool for future use in clinical trials and standard practice.
Overall Survival according to HCT-specific Comorbidity Index Overall Survival according to HCT-specific Comorbidity Index
Fanconi Anemia D2 Protein Contributes to BCR/ABL-Mediated Transformation of Hematopoietic Cells.
Authors: Koptyra Mateusz , Houghtaling Scott , Grompe Marcus , Skorski Tomasz 
Published: October 2019

Abstract
Homologous recombination (HR), involving RAD51 protein, plays an important role in the response of BCR/ABL-positive leukemia cells to numerous DNA double-strand breaks (DSBs) induced by reactive oxygen species (ROS) or genotoxic treatment. Fanconi D2 protein (FANCD2), a member of the Fanconi protein family, is monoubiquitinated on K561 and phosphorylated by ATM on S222 in response to DSBs. The K561 monoubiquitinated form of FANCD2 interacts with RAD51 during HR, and phosphorylation of FANCD2 on S222 is important for activation of S phase checkpoint in response to DNA damage. Our studies detected an enhanced interaction between RAD51 and FANCD2 in BCR/ABL-positive leukemia cells in comparison to normal counterparts. In addition, although the expression of FANCD2 was stimulated by BCR/ABL and growth factors, higher levels of FANCD2 monoubiquitination was detectable in CML patient cells at chronic phase and in blast crisis, and in BCR/ABL-transformed cells in comparison to non-transformed cells. This effect was reversed after inhibition of BCR/ABL kinase with STI571. Therefore, monoubiquitination of FANCD2 may play a role in BCR/ABL-mediated leukemogenesis. BCR/ABL kinase displayed an impaired transformation potential in FANCD2-/- murine bone marrow cells in comparison to +/+ counterparts. In addition, expression of BCR/ABL kinase, but not the kinase-deficient K1172R mutant, inhibited the proliferation rate of FANCD2-/- human lymphoblast cell line. Growth ability of BCR/ABL-positive FANCD2-/- cells could be rescued by co-expression of the wild-type and S222A mutant of FANCD2, but not the K561R mutant. This observation suggested that K561 monoubiquitination, but not S222 phosphorylation might play an important role in BCR/ABL-mediated transformation. Since BCR/ABL cells employ RAD51-dependent HR to repair numerous DSBs induced by ROS, elevated expression of monoubiquitinated FANCD2 may facilitate this process. This hypothesis is supported by the observation that BCR/ABL-positive FANCD2-/- cells accumulate more DNA damage than +/+ counterparts as indicated by enzymatic assays converting oxidative DNA lesions into gaps detectable by comet assay. In addition, enhanced oxidative DNA damage in BCR/ABL-positive FANCD2-/- cells produced a variety of DNA lesions including abasic sites, and single- and double-strand breaks assessed by neutral comet assay. Moreover, BCR/ABL-positive FANCD2-/- cells accumulated higher numbers of DSBs detected by γ-H2AX immunostaining and displayed discrete apoptosis. In conclusion we hypothesize that monoubiquitination of FANCD2 may play a role in the initial steps of BCR/ABL dependent leukemogenesis, probably due to its ability to interact with RAD51 and facilitate HR repair of an excess of spontaneous DSBs induced by ROS.
MDM2 Dependant Induction of Error Prone DNA Polymerase beta by EBV-LMP1 Oncoprotein.
Authors: Nathalie Faumont , Christophe Le Clorennec , Pierre Teira , Ibtissam Youlyouz-Marfak , Christine Mazeres , Sebastien Hoffmann Jean , Georges Delsol , Jean Feuillard , Fabienne Meggetto 
Published: October 2019

Abstract
Epstein Barr Virus (EBV) is able to immortalise B lymphocytes in vitro and is associated with several human tumours, such as Hodgkin’s disease (HD), suggesting that this virus could play a role in the initiation of the tumoural process. The LMP1 protein of EBV, coded by the LMP1-BNLF1 gene, was rapidly suspected to be involved in this process due to its oncogenic characteristics. LMP1 functions as a constitutively activated member of the tumour necrosis factor receptor superfamily, and activates trough its carboxyterminal cytoplasmic region several signalling pathways, including NF-kappa B. In order to identify new LMP1 target genes, we studied the transcriptome of the B lymphoid Ba/F3 cell line stably transfected with LMP1-BNLF1 genes from HD Reed-Sternberg tumour cells as well as from normal cells, using the cDNA microarray technique.
We show that the expression of several genes was modulated by the LMP1 protein, independently of the cellular origin of the LMP1 protein. Among overexpressed genes, we focused on the DNA polymerase-beta (beta-pol) gene. To confirm and extend this cDNA result, we demonstrated in different cell lines that the beta-pol protein expression was under the control of LMP1. In vivo and in vitro functional tests showed that LMP1 induced overexpression of beta-pol paralleled its increase in activity.
Regulatory sequences of the human beta-pol gene includes three motives that match the Sp1 factor binding site motive. LMP1 did not modulate Sp1 RNA neither protein expression. By contrast, we observed that MDM2 and beta-pol RNA expression were increased in parallel by LMP1, an effect trigerred by the carboxyterminal region of LMP1. Then, we demonstrated that MDM2 protein overexpression was associated with increased beta-pol protein expression.
Since MDM2 promoter is regulated by NF-kappaB, we investigated whether up-regulation of MDM2 and beta-pol were NF-kappaB pathway dependant. We show that inhibition of NF-kappaB by overexpression of an I-kappaB dominant negative down-regulated both MDM2 and beta-pol expression.
In conclusion, we demonstrated that the oncoprotein LMP1 regulates overexpression of beta-pol through NF-kappaB signaling pathway and the MDM2 protein, and that MDM2 was itself regulated by NF-kappaB. Beta-pol is the more error prone of all the known eucaryotic DNA polymerases. Elevated expression of beta-pol, an event found in many human tumours, has been shown to generate a mutator phenotype which is extensively associated with cancers. Our work leads to the hypothesis that LMP1 protein could play a role in the genetic instability in EBV associated malignant diseases such as HD through over-expression of beta-pol.
Differential Expression of the Homeobox Gene MIXL1 in Non Hodgkin (NHL) and Hodgkin Lymphomas (HL).
Authors: Drakos Elias , Rassidakis George Z. , Guo Wei , Medeiros L. Jeffrey , Nagarajan Lalitha 
Published: October 2019

Abstract
The gene MIXL1 (Mix1 homeobox-like) encodes a paired class homeobox transcription factor involved in early hematopoietic specification during embryogenesis. Previous studies have shown that MIXL1 gene is expressed in hematopoietic cells during adult life (Guo et al. Blood100;1;89–96, 2002). Furthermore 5′ MIXL1 sequences are a target of retroviral insertion in murine T-cell lymphoma (http:RTCGD.ncifcrf.gov), suggesting a selection advantage for aberrant expression of this gene. However, the status of MIXL1 expression in human lymphomas has not been examined. Using a highly specific antibody, we assessed for MIXL1 protein expression in 14 lymphoma cell lines (9 B-cell and 5 T-cell) by immunobloting. MIXL1 was detected predominantly in nuclear extracts of lysates of all cell lines tested, although at a variable level. We also assessed for MIXL1 protein expression in 126 B-cell and 21 T-cell NHLs of various types, as well as 14 Hodgkin lymphomas using immunohistochemical methods. The results of the immunohistochemical studies are summarized in table 1. Once again, MIXL1 immunoreactivity was primarily nuclear in the tumor cells. Based on distribution data (histogram), a 50% cutoff was selected for high versus low MIXL1 expression. Among B-cell tumors, high expression levels of MIXL1 protein were more frequently detected in high-grade NHL and HL compared with low/intermediate grade NHL (p<0.0001, chi-square test). As a continuous variable, the percentage of MIXL1-positive tumor cells was also significantly higher in high-grade B-cell NHL and HL compared with low/intermediate grade NHL (p<0.0001, Kruskal Wallis test). All Hodgkin lymphomas expressed high levels of MIXL1 with 60% to 100% of neoplastic cells being positive for MIXL1. Most T-cell NHLs also expressed high levels of MIXL1. In contrast, most low/intermediate-grade B-cell NHL and multiple myelomas expressed low levels of MIXL1. Frequent overexpression of MIXL1 gene product in most high-grade B-cell NHLs, HL and T-cell NHLs suggests that aberrant expression of MIXL1 may play a role in proliferation, block of differentiation or both.
Table 1. HL (n=14) B-NHL (n =126) T-NHL (n =21) N (%) Low/intermediate grade N (%) N (%) Classical HL 12/12(100%) Chronic lymphocytic leukemia /small lymphocytic lymphoma 0/8 (0% T-precursor lymphoblastic leukemia/lymphoma 2/2 (0%) Nodular lymphocyte predominance HL 2/2 (100%) MALT-lymphoma 0/8 (0%) Mycosis fungoides/Sezary syndrome 2/2 (0%) Follicular lymphoma 9/24 (38%) Extranodal NK/T-cell lymphoma, nasal type 3/3 (100%) Mantle cell lymphoma 5/34 (15%) Peripheral T-cell lymphoma, unspecified 6/9 (66% High grade Anaplastic large cell lymphoma 5/5 (100%) B-precursor lymphoblastic leukemia/lymphoma 1/3 (33%) Burkitt lymphoma/leukemia 2/2 (100%) Diffuse large B-cell lymphoma 30/31 (97%) Plasma cell myeloma/plasmacytoma 0/16 (0%)
Management of HIV Associated Thrombotic Thrombocytopenic Purpura.
Authors: Scully Marie , Miller Robert , Cohen Hannah , Roedling Sherie , Edwards Simon , Machin Samuel J. 
Published: October 2019

Abstract
Thrombotic thrombocytopenic purpura (TTP) is a life threatening disorder characterised by thrombocytopenia and microangiopathic haemolytic anaemia associated with a deficiency in ADAMTS 13. Human Immunodeficiency Virus (HIV) remains an important infectious cause of TTP. We report the characteristics and treatment of patients presenting over a 6.5 year period to a tertiary referral centre. Cases: 9 patients, 8 female, 1 male, all heterosexual Africans. In 5 of the patients, TTP presentation was associated with the initial diagnosis of HIV. Only 1 patient was on highly active anti-retroviral therapy (HAART) at diagnosis. Median haemoglobin on admission was 6.4 (range 4.6–7.2) g/dl, platelet count-12 (range 5–53) X 109/L, lactate dehydrogenase (LDH) 1401 (range 914-2532) IU/L. In 6 patients there were neurological features at presentation, from headache, to transient ischaemic attacks and coma<INS cite=mailto:HCOHEN dateTime=2005-07-29T11:48>;</INS>6 patients had renal impairment, median creatinine 127 (range 63–418) μmol/L. Notably, HIV associated TTP was not a late onset disorder associated with poor CD 4 counts and poor prognosis. Median CD 4 counts were 170 (range 70–400) cells/μl, with a median viral load 74,400 copies/ml. ADAMTS 13 activity ranged from <5%–50% (normal range 66–126%) with no evidence of inhibitor by mixing studies using collagen binding assay. All patients received single volume plasma exchange<INS cite=mailto:HCOHEN dateTime=2005-07-29T11:50> </INS>(PEX) with cryosupernatant, median 8 (range 3-16) until remission and started HAART as soon as the diagnosis was made. Two patients received fresh frozen plasma (FFP) replacement for PEX for refractory TTP: one had coma and suboptimal treatment pre-referral, the second patient had an initial speedy recovery, but developed septicaemia. Two patients stopped their anti-retroviral therapy and had a TTP relapse with<INS cite=mailto:HCOHEN dateTime=2005-07-29T11:51>in</INS> 3 months. However, recommencement of HAART plus 3 and 6 PEX’s respectively was associated with remission. Six of the patients remain in remission from TTP at 6–74 months; 2 were lost to follow-up and 1 died 18 months later from renal failure, unrelated to TTP. These data suggest HIV status should be checked in all patients presenting with acute TTP as it accounted for 12.5% of all acute TTP admissions in our cohort. All HIV associated TTP patients were black African heterosexual<INS cite=mailto:HCOHEN dateTime=2005-07-29T11:56>s</INS>. Treatment with PEX and HAART was associated with speedy remission of TTP. In patients who stopped HAART, TTP relapsed within 3 months. The abscence of antibody to ADAMTS 13 suggests a possible direct effect of the retrovirus on endothelium. With HAART and a decrease in viral load, ADAMTS 13 activity increases as patients achieve remission.
Adoptive Transfer of Nf1+/− Bone Marrow Is Sufficient for Neurofibroma Progression in Krox20;Nf1flox/flox Mice.
Authors: Yang Fengchun 
Published: October 2019

Abstract
Mutations in the NF1 tumor suppressor gene cause neurofibromatosis type 1 (NF1), a GTPase activating protein for Ras called neurofibromin. NF1 is a genetic disorder that affects approximately 250,000 individuals in the US, Europe, and Japan alone. Neurofibromas, the hallmark of NF1, are complex tumors characterized by tumorigenic Schwann cells, neoangiogenesis, fibrosis, and degranulating mast cells. Studies in experimental models have emphasized the role of inflammatory cells in altering the microenvironment and facilitating malignant outgrowth. Similarly, Parada (Science, 2002) found that nullizygosity of Nf1 in Schwann cells of conditional knockout mice (Krox20;Nf1flox/flox) was necessary but not sufficient for neurofibroma formation and haploinsufficiency of Nf1 in lineages within the tumor microenvironment was required for neurofibroma progression. We previously provided the first genetic, cellular, and biochemical evidence that haploinsufficiency of Nf1 alters Ras activity and cell fates in mast cells (JEM, 2000, 2001) and identified a mechanism underlying the recruitment of mast cells to tumorigenic Schwann cells (JCI 2003). However, it remains unclear whether Nf1 +/− bone marrow derived hematopoietic cells can directly contribute to neurofibroma formation in vivo. To address this question, Nf1+/− or wildtype (WT) EGFP+ bone marrow (BM) was adoptively transferred into lethally irradiated Krox20;Nf1flox/flox mice and cohorts were followed prospectively for tumor formation using positron emission tomography and computerized axial tomography. Mice transplanted with Nf1+/− but not WT BM developed progressive enlargement of the trigeminal nerve, dorsal root ganglia, peripheral nerves, and motor paralysis similar to Krox20;Nf1flox/− mice that are haploinsufficient at Nf1 in all lineages of the tumor microenvironment. Postmortem analysis revealed that Krox20;Nf1flox/flox mice transplanted with Nf1+/− BM had cellular neurofibromas containing Schwann cells, fibroblasts, blood vessels and mast cells, which closely resembled the cellular architecture of human neurofibromas. Mice transplanted with WT BM did not develop neurofibromas. These studies establish that recruitment of Nf1 +/− BM derived cells to the neurofibroma microenvironment is directly linked to neurofibroma formation and progression. Given our observations, therapies which prevent both the recruitment and the tumor promoting functions of Nf1 +/− hematopoietic cells in neurofibroma formation are currently being tested in vivo as pre-clinical trials.