bluuebunny/crossref_metadata_2025_split huggingface dataset:
Biochemical and Cellular Characterization of the Novel Proteasome Inhibitor PR-171.
Authors: Demo Susan D. , Buchholz Tonia J. , Laidig Guy J. , Parlati Francesco , Shenk Kevin D. , Smyth Mark S. , Sullivan Carol M. , Sun Congcong M. , Vallone Marcy K. , Woo Tina , Bennett Mark K. 
Published: October 2019

Abstract
Recent clinical studies have identified the proteasome as an important therapeutic target for hematologic malignances. The proteasome inhibitor, bortezomib, has been approved for the treatment of relapsed or refractory multiple myeloma and ongoing clinical trials suggests a potential benefit for the treatment of Non-Hodgkin’s lymphoma. PR-171 is a novel epoxomicin derivative that is a potent and irreversible inhibitor of the human proteasome. It inhibits the chymotrypsin-like activity of purified human 20S proteasome with a kinact/Ki of 34,000 M−1s−1 and is >300-fold selective over the other proteasome catalytic activities. In addition, PR-171 has been found to exhibit minimal activity in a broad diversity panel of biochemical assays that includes 67 receptor/ligand and 37 enzyme assays. PR-171 retains its potency for inhibition of the proteasome chymotrypsin-like activity in mammalian cells displaying IC50 values <10 nM in multiple tumor cell lines. Although PR-171 is a covalent irreversible inhibitor, proteasome activity in cells recovers with a t1/2 of approximately 24 hr after removal of the compound. This recovery is likely due to induction of de novo synthesis since other proteasome inhibitors have been shown to promote transcription of multiple proteasome subunit genes. The cellular consequences of proteasome inhibition by PR-171 include accumulation of polyubiquitinated proteins, cell cycle arrest, and induction of apoptosis. Potent cytotoxic activity of PR-171 is observed across a broad panel of human tumor cell lines (IC50 range: 2–40 nM). Pulsatile exposure studies designed to mimic the anticipated drug exposure in vivo have demonstrated that cytotoxicity is dependent upon the magnitude and duration of proteasome inhibition. These studies have also shown that hematological tumor lines are most sensitive to brief PR-171 exposure, with solid tumor lines exhibiting intermediate sensitivity and non-transformed cells being the least sensitive to such treatment. Finally, PR-171 has been found to retain its cytotoxic potential on cells made resistant to bortezomib in vitro. These studies motivate the clinical investigation of PR-171 in hematological malignances.
The Role of Stromal Cells in Multiple Myeloma: Actors or Spectators?.
Authors: Corso A. , Ferretti E. , Gallì A. , Tenore A. M. , Pascutto C. , Zappasodi P. , Mangiacavalli S. , Varettoni M. , Rusconi C. , Lazzarino M. 
Published: October 2019

Abstract
Multiple myeloma (MM) is a B cell neoplasia characterized by an accumulation of clonal plasma cells (PCs) in the bone marrow (BM). The growth and survival of MM plasma cells is regulated by their network with the microenvironment, mainly with the stromal cells. However, although bone marrow stromal cells have been shown to take part in the pathogenesis of the disease, it is still unknown whether these cells play an active or passive role. Namely, whether normal stromal cells simply supply the demand of MM plasma cells, or, during the course of the disease, they acquire abnormal characteristics becoming pathological. To address this question, we designed an in vitro co-culture model in which PCs isolated by immuno-magnetic separation from MGUS and MM patients are crossed with BM stromal cells (BMSCs) derived from MGUS and MM patients. As a result, four type of co-cultures were obtained: MM-BMSCs/MM-PCs, MM-BMSCs/MGUS-PCs, MGUS-BMSCs/MM-PCs, MGUS-BMSCs/MGUS-PCs. After two days of co-culture in a serum free medium, we evaluated the survival of MM-PCs or MGUS-PCs for each combination. We also quantified by ELISA assays in the supernatants of the same cultures, the level of several growth factors (IL-6, IL-8, VEGF, MIP-1a, MIP-1b, RANTES, MCP-1, TGF-b, SDF-1) to evaluate the possible influence of these cytokines on plasma cells. Multivariate general linear models were applied to compare survival in the different combinations of BMSCs and PCs, also accounting for the various growth factors. MM-BMSCs showed to support the survival of both MM-PCs and MGUS-PCs significantly better than MGUS-BMSCs (p=0.0007). However, in the combination MGUS-PCs/MGUS-BMSCs plasma cells survived statistically better than in that MM-PCs/MGUS-BMSCs (p=0.00003). As regards the cytokines, IL-6, IL-8, VEGF, MIP-1a, MIP-1b, and RANTES did not show to be significantly associated with plasma cell survival in all settings. TGF-B and SDF-1 levels were significantly associated with better survival of both MM-PCs and MGUS-PCs when cultured with MM-BMSCs compared to MGUS-BMSCs (p=0.0001 and p=0.038, respectively), while MCP-1 was significantly associated with reduced survival of MM-PCs and MGUS-PCs in the same setting (p=0.006). In conclusion, these data favours the concept that the behaviour of stromal cells may change during the transition from the condition of MGUS to the overt state of myeloma, evolving from a simple role of a spectator to that of an actor. It also appears that overt MM plasma cells have the highest need for cytokine supply and therefore are more dependent on BMSCs activity.
Disparities in Criteria for Initiating Chelation Therapy for Iron Overload in Patients with Myelodysplastic Syndromes (MDS).
Authors: Bennett John M. , List Alan F. ,   for the Myelodysplastic Syndromes Foundation Inc.
Published: October 2019

Abstract
In the majority of MDS cases, isolated anemia is the only clinical evidence of impaired hematopoiesis at the time of MDS diagnosis. It is estimated that >40% of MDS patients require regular red blood cell transfusions during some stage of their disease. Findings from The MDS Foundation’s 2004–2005 Practices & Treatment Survey indicate that a substantial proportion of MDS patients in all International Prognostic Scoring System (IPSS) risk groups are transfusion-dependent: Low, 30%; Intermediate-1, 50%; Intermediate-2, 70%; High, 90% (median values). Seventy (38 US and 22 international) of the MDS Foundation’s102 Centers of Excellence responded to the survey as of July 1, 2005. Survey responses revealed that an average of 30% of transfusion-dependent patients receive parenteral iron chelation therapy (median, 20%; range: 3–100%) and that the criteria for initiating chelation therapy are not uniform. The number of transfusions was reported as a determining criterion by 46% of respondents, with a mean number of 35 transfusions (median, 30; range: 4–100). 15% of respondents indicated at the number of transfusions was the sole criterion used. Serum ferritin levels were reported as a determining criterion by 57% of respondents, with the following cutoff values:
Ferritin concentrations for initiating chelation therapy *% respondents using this cutoff as criterion >1000 ng/mL 41%* >1500 ng/mL 11% >2000 ng/mL 37% Other (>3000, unspecified) 9%
32% of respondents indicated that serum ferritin was the sole criterion used. (Irrespective of chelation therapy, 92% indicated that they monitored ferritin levels in transfusion-dependent patients.) Other criteria used to determine start of chelation therapy included age/life expectancy, MDS subtype, clinical signs of hemochromatosis, quantitative CT liver iron estimation, liver function, transferrin saturation >50%, BMT, anticipated chronic transfusion need, and logistical issues and insurance coverage. A combination of criteria was reported to be used by 38% of respondents. The majority of respondents (90%) reported that they would increase the number of patients on chelation therapy if an oral agent were available. Although the decision for initiating chelation therapy in transfusion-dependent anemic MDS patients is individualized because of the heterogeneous patient population, this data analysis suggests a need for the standardization of select criteria (e.g., number of transfusions, serum ferritin).
Heparin Synergistically Enhances Interleukin-11 Signaling through Upregulation of the Map Kinase Pathway.
Authors: Rajgopal Raghav , Butcher Martin , Walton Kimberly J. , Shaughnessy Stephen G. 
Published: October 2019

Abstract
Using an animal model of heparin-induced osteoporosis we previously demonstrated that heparin causes bone loss, in part, by increasing osteoclast number and activity. In an effort to account for these findings, we examined the effect of heparin on osteoclast formation in vitro. We found that while heparin alone had no effect, it was able to synergistically enhance Interleukin-11 (IL-11) - induced STAT3 (Signal Transducer and Activator of Transcription) activation and thus, increase both gp130 expression and osteoclast formation. In the present study, we examine the effect of various serine kinase inhibitors on heparin’s ability to synergistically enhance both STAT3 activation and gp130 expression. As assessed by electrophoretic mobility shift assays (EMSAs), treatment of murine calvaria cells with heparin was found to increase IL-11-induced STAT3-DNA complex formation by 2 fold. Inhibition of the c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein (MAP) kinase, or the phosphatidylinositol 3-kinase (PI3-K) pathway had no effect on heparin’s ability to promote STAT3-DNA complex formation. In contrast, the mitogen-activated protein kinase kinase (MEK) inhibitor, PD098059, completely abolished the ability of heparin to act synergistically with IL-11. Similar results were obtained when northern blot analysis was used to assess heparin’s ability to enhance IL-11-induced gp130 expression. In an attempt to resolve the mechanism by which this was occurring, we examined the ability of heparin to influence STAT3 Ser727 phosphorylation in the presence or absence of IL-11. Heparin alone was found to have no effect on Ser727 phosphorylation, nor did heparin alter the phosphorylation status of Ser727 in the presence of IL-11. Heparin was, however, found to increase ERK1/2 (extracellular signal regulated kinase) activation in both a time and dose-dependent manner. When taken together, these findings suggest that heparin enhances IL-11-induced STAT3 activation and thus, gp130 expression, by a mechanism that is independent of STAT3 Ser727 phosphorylation, but which involves upregulation of the MAP Kinase pathway.
Marker Expression in Peripheral T-Cell Lymphoma Unspecified: Proposal of a Clinical-Pathologic Prognostic Score.
Authors: Piccaluga Pier Paolo , Went Philip , Agostinelli Claudio , Gallamini Andrea , Ascani Stefano , Sabattini Elena , Bacci Francesco , Falini Brunangelo , Motta Teresio , Paulli Marco , Artusi Tullio , Zinzani Pier Luigi , Baccarani Michele , Pileri Stefano A. ,   on Behalf of the Intergruppo Italiano Linfomi
Published: October 2019

Abstract
Peripheral T-cell lymphoma unspecified (PTCL/U) represents the commonest form of T-cell tumor in Western Countries according to the WHO Classification. So far, no concrete attempts have been made in order to apply a wide panel of markers to a large series of PTCLs and to assess the impact of phenotype on prognosis and survival. We then studied the protein expression and outcome of 148 PTCL/U cases, along with 45 tumors of the AILD type, utilizing highly standardized high-throughput technology.
Tissue micro-arrays corresponding to the above mentioned cases were constructed and analyzed with a panel of 18 commercially available markers. In 93 patients with PTCLs/U clinical data were available and were matched with the protein expression profile. Interestingly, most of these patients had been included in a previous study that proposed a prognostic index for PTCL/U (PIT) (Gallamini et al. Blood2004, 103:2474–9).
An aberrant phenotype with frequent loss of CD5 and/or CD7 was typical for all PTCLs, irrespectively of the subtype (unspecified or AILD-type). CD20 and CD15 were rarely aberrantly expressed, at times simultaneously with CD30. EBER positivity and CD15 expression emerged as adverse prognostic factors, while CD56 and CD57 were unremarkable. Among PTCLs/U, the proliferation-associated protein Ki-67 was found to be prognostically relevant and was then integrated in a new prognostic score, including age (>60 years), high serum lactate dehydrogenase, poor performance status, and Ki-67 3 80%. Such score was associated with the overall survival (p<0.0001) and was more a powerful predictor than PIT that - however - maintained its relevance.
Our retrospective analysis shows a wide range of protein expression in PTCLs and candidates a new prognostic index. The latter represents one of the first examples of mixed score (including patient- and tumor-specific factors) applied to malignant lymphomas and may be the basis for future prospective therapeutic trials.
Autologous Non-Myeloablative Hematopoietic Stem Cell Transplantation for Diffuse Scleroderma.
Authors: Oyama Yu , Statkute Laisvyde , Barr Walter , Traynor Ann , Burt Richard 
Published: October 2019

Abstract
Background &Aims Scleroderma is an immune mediated vasculopathy of unknown etiology. There is no known therapy that could alter the natural course of the disease. Patients with diffuse cutaneous disease or visceral involvement has an overall 10-year survival of 35 to 68%. Autologous hematopoietic stem cell transplantation (HSCT) with myeloablative regimen has been performed, but with substantial mortality and morbidity especially pulmonary and renal toxicity. We have performed autologous HSCT utilizing non-myeloablative, but lymphoablative conditioning.
Methods
We conducted a phase 1 HSCT study in 9 patients with diffuse scleroderma with poor clinical features. Candidates were less than 65 years old with Rodman score of more than 14 or diffusion capacity less than 80%, interstitial lung disease, elevated ESR, renal involvement, or abnormal electrocardiogram. Patients with pulmonary hypertension (systolic pressure > 45 mm Hg) were excluded. Peripheral blood stem cells were mobilized with cyclophosphamide and granulocyte colony-stimulating factor. The graft was not manipulated. The conditioning regimen consisted of 200mg/kg cyclophosphamide (CY), and 7.5mg/kg rabbit antithymocyte globulin (ATG).
Results
The procedure was well tolerated with anticipated cytopenias, neutropenic fever, and fluid overload that were easy to manage and without mortality. The median days for neutrophil and platelet engraftment were 8 (range 7–9) and 8 (range 0–10), respectively. The median infused CD34+ and CD3+ cell counts were 8.31 x 106/kg (range 2.35–14.7) and 2.03 x 108/kg (range 0.41–6.83), respectively. There was a marked improvement of skin score in all subjects, whereas cardiac (ejection fraction, pulmonary arterial pressure), pulmonary function (DLCO) and renal function (creatinine) remained stable. One patient with advanced disease with poor performance status died 2 years after the transplant with progressive disease. After median follow-up of 20 months (range 5–32), the overall survival is 89% (eight out of nine) and progression-free survival with continuing improvement is 67% (six out of nine. 2 patients developed recurrence of skin tightness without compromising organ function. Both of them were placed on mycophenolate mofetil with gradual improvement of skin tightness.
Conclusion
Autologous HSCT with CY/ATG is safe and is effective. A randomized study (ASSIST: American Scleroderma Stem Cell vs. Immune Suppression Trial), comparing HSCT to intravenous pulse cyclophosphamide is now enrolling patients.
Figure Figure
Intratumoral CD4+CD25+ Regulatory T-Cell-Mediated Suppression of Infiltrating CD4+ T-Cells in B-Cell Non-Hodgkin Lymphoma.
Authors: Yang Zhi-Zhang , Novak Anne J. , Stenson Mary J. , Witzig Thomas E. , Ansell Stephen M. 
Published: October 2019

Abstract
Background: Non-Hodgkin lymphomas (NHL) are increasing in incidence and are now the fifth most common tumor diagnosed each year in the United States. Most NHLs are of B-cell origin but the tumor tissue is variably infiltrated with T-cells. Our group has shown in diffuse B-cell large cell lymphoma, that a high number of intratumoral CD4+ T-cells predicts a better overall survival. It has been shown that recently-characterized CD4+CD25+ regulatory T-cells (Treg cells) played an important role in the mediation of anti-tumor immunity. However, there is no data on the role of intratumoral Treg cells in suppression of autologous infiltrating CD4+ T-cells in B-cell NHL.
Goal: To investigate the effect of intratumoral Treg cells on the proliferation of tumor-infiltrating CD4+CD25- T-cells, to determine the underlying mechanism of the T-cell suppression, and evaluate the role that malignant B-cells may play in the recruitment of Treg cells to the site of B-cell NHL.
Results: We identified a subset of CD4+CD25+ T-cells over-represented in biopsy specimens of B-cell NHL (these cells comprise 17% of cells in lymphoma biopsies, compared 7% of peripheral blood mononuclear cells, 12% of cells in inflammatory tonsil and 6% of cells in tumor free lymph nodes; p-value =0.001). These CD4+CD25+ T-cells are memory-like T-cells (CD45RO+ and CD45RA−) and express high levels of CTLA-4 and Foxp3 when compared to autologous tumor-infiltrating CD4+CD25- T-cells. Importantly, these CD4+CD25+ T-cells displayed the ability to suppress the proliferation and cytokine (IFN-g and IL-4) production of tumor-infiltrating CD4+CD25- T-cells in response to PHA stimulation. Treatment with anti-B7-H1 antibody or PD-1 fusion protein enhanced the proliferation of infiltrating CD4+CD25- T-cells when co-cultured with intratumoral CD4+CD25+ T-cells. Our results suggest that interaction between B7-H1 and PD-1 accounts for about 30% of intratumoral Treg cell-mediated inhibition of autologous infiltrating CD4+CD25- T-cells in tumor sites of B-cell NHL. Lastly, we found that CCL22 secreted by lymphoma B-cells is involved in the chemotaxis and migration of intratumoral CD4+CD25+ T-cells which express chemokine receptor CCR4, but not CCR8.
Conclusion: Our results suggest that tumor microenvironmental CD4+CD25+ regulatory T-cells are important regulators of tumor immunity and that these cells are recruited to the area of lymphoma involvement by the malignant B-cells.
Liver Hepcidin mRNA Is down Regulated at High Liver Iron Burdens and up Regulated in HCV Infection in Thalassaemia.
Authors: Farrukh Shah T. , Monica Mascarenhas , Roozina Rafique , Kaila Srai S. , John Porter B. 
Published: October 2019

Abstract
Rationale. Hepcidin is an important negative regulator of iron absorption from the gut and iron release from macrophages. Synthesis is increased by inflammation and decreased in iron deficiency. Iron overload conditions might be predicted to up-regulate hepcidin synthesis, thereby decreasing dietary iron absorption. However in adult onset herediditary hemochromatosis, there is paradoxical down regulation of liver hepcidin mRNA (Bridle et al.,Lancet 2003). Urinary hepcidin is decreased thalassaemia major (Papanikolaou et al., Blood 2005) but expression of mRNA has not been described. However, hepcidin mRNA may be increased in iron overloaded in patients with chronic HCV infection (Aoki, et al., J Clin Gasteroenterol 2005).
Methodology. mRNA was extracted on 20 liver biopsies from thalassaemia patients and RT PCR performed from cDNA as described by Leung et al., Blood 2005. 14 biopsies were obtained from thalassaemia major (TM) patients on regular transfusion programmes and the remaining 6 from thalassaemia intermedia (TI) patients, 2 of whom were not currently receiving transfusion, 3 were on exchange transfusion regimes and one was on 6–8 weekly top up transfusions. All samples were calculated as the hepcidin/actin ratio and were measured simultaneously with liver biopsies obtained from healthy control subjects at the time of surgery. Hepcidin mRNA was then expressed relative to control values from normal liver tissue in healthy control subjects.
Results. Mean hepcidin mRNA was significantly lower in patients with liver iron concentrations >10mg/g/dw compared to those with lower burdens (85 vs 131%, p=0.05). Hepcidin mRNA did not vary significantly with anaemia, with ferritin > 3000ng/ml or <1500ng/ml (p=0.99). Hepcidin mRNA was not significantly different between the 2 thalassemia groups; TI (123 ± 47%) compared to (110 ± 73%)TM. 3 patients with high transaminase levels (>80 iu/ml) all had low hepcidin mRNA levels (77 ± 16 %) and increased levels of fibrosis compared to those with lower transaminase levels (106 ± 55%) and only mild fibrosis. 4 patients who were HCV antibody positive but with only modestly elevated transaminase levels (<160iu/ml) and mild/moderate fibrosis had raised hepcidin mRNA (163 + 70 %) p=0.02. There was no difference in the HCV positive and HCV negative groups for ferritin (2530 ng/ml: 2483 ng/ml), ALT (61 iu/l :53 iu/l) or liver iron (7.5 mg/g/dw versus 8.2 mg/g/dw).
Conclusions. This suggests a critical level of iron burden (LIC) above which hepcidin expression becomes dysregulated, failing to increase with further elevations in the iron burden and paradoxically decreases. This could lead to inappropriately high iron absorption in heavily iron-loaded patients. Of other interacting factors affecting hepcidin synthesis, this study suggests that active HCV may increase hepcidin synthesis.
Klf12 Is Required for Vessel Organization in Zebrafish Embryos.
Authors: Perkins Andrew C. , Gunaratnam Dinushka , Gardiner Melissa R. , Robinson Kathleen C. 
Published: October 2019

Abstract
The process of forming a lumenised vessel from an angioblast cord is a crucial part of both vasculogenesis and angiogenesis. The Krüppel like factors (klfs) are a family of zinc finger transcription factors which play important roles in many aspects of differentiation. Klf2 knockout mice die in utero from haemorrhaging due to arterial wall defects. Due to the similarity of vascular and haematopoietic systems to those of mammals, the zebrafish was chosen as a model in which to study the function of klf12, the only zebrafish representative of the repressor subfamily of mammalian klfs which includes klf3, klf8 and klf12. Klf12 is first detected by WISH at 12 somites in the lateral plate mesoderm (LPM) and continues in these cells as they from the ICM at 18–22hpf, the site of vasculogenesis and haematopoiesis. From 24hpf klf12 is expressed in short stripes extending from the ICM dorsally towards the notochord. This expression pattern is similar but not identical to that of fli1 and flk1, two vascular markers. Expression subsequently decreases and is absent by 30hpf.
Targeted knockdown of klf12 by translantion-inhibiting and splicing morpholinos (MOs) produces a vascular defect. At 24hpf morphants display correct expression of primitive blood, kidney and vascular markers such as gata1, βE3globin, biklf, pax2.1 and fli1. However, circulation is absent in 65% of embryos at 48 hpf and reduced in most, as seen by both brightfield microscopy and fluorescent microbead microangiography. Embryos also display a slower heartbeat, pericardial oedema and oedema over the yolk/duct of Cuvier, all likely to be secondary effects due to the circulatory defect. Injection of klf12 MOs into fli1-eGFP transgenic embryos reveals correct differentiation of endothelial cells, but disorganised angiogenesis.
In summary, klf12 morphants display correct specification of angioblasts and differentiation of endothelial cells but a defect in tubulogenesis of endothelial cords.