bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic Dissection of Cooperating Mutations in BXH-2 Acute Myeloid Leukemia with and without Nf1 Gene Mutation.
Authors: Yin Bin , Walrath Jessica , Shannon Kevin M. , Wallace Margaret R. , Largaespada David A. 
Published: April 2020

Abstract
Loss of the NF1 (Neurofibromatosis Type 1) gene, a tumor suppressor, can cause myeloid diseases juvenile myelomonocytic leukemia (JMML), monosomy 7 syndrome (Mo7), and acute myeloid leukemia (AML). However, using knockout mice, it has been shown that loss of Nf1 expression in hematopoietic cells, by itself, does not lead to aggressive leukemia resulting instead in a relatively indolent myeloproliferative disease. Murine Leukemia Virus (MuLV) insertional mutagenesis in BXH-2 mice provides a model to dissect genetic alterations in AML. We have profiled proviral insertions in BXH-2 AML which do or do not have corresponding loss-of-function of Nf1. 197 PIS (68 from 25 Nf1-wild type AML and 129 from 55 Nf1-defective AML) were isolated. Nf1-defective AML were obtained from BXH-2 AMLs with proviral insertions into the endogenous Nf1 gene and AML that developed in leukemia-prone, heterozygous Nf1+/− BXH-2 mice. These latter AMLs develop faster than wild-type BXH-2 AMLs and show Nf1 gene LOH or proviral insertion into the wild-type Nf1 allele. These analyses led to 37 common proviral insertion sites (CIS), 13 of which have not been reported previously. Several of the CIS (including Lmo2, Cmyb, Meis1, Bcl11a, Spred2, Def8, Edg3, Hoxa9, and a novel Krab domain-zinc finger gene) were found repeatedly among the Nf1-defective group of AML. Expression of most could be detected in human JMML and CMML by RT-PCR, including BCL11A. Importantly, among the CIS we detected, PIS targeting Bcl11a were significantly enriched (p &lt; 0.05) in Nf1-defective leukemia. Retroviral expression vectors for Bcl11a have been constructed and transduced into an immortalized Nf1-/- null myeloblast cell line. Growth assays show that the cumulative cell number of FACS-sorted Bcl11a-Nf1-/- cells increase by ~2.5 fold that of controls. BXH-2 provides a powerful genetic system to dissect Nf1-cooperating genetic events in tumorigenesis. Mutations at several novel common integration sites could be involved in development or progression of leukemia with NF1 gene inactivation. This work was supported by the National Cancer Institute (U01-CA84221-05) and the American Cancer Society (RPG LIB-106632) to DAL and by National Cancer Institute (R01 CA92095) and U.S. Dept. of Defense (DAMD17-97-1-7339) to MRW.
Induction of G1 Arrest and Apoptosis in a Conditional Expression Model of AML1/ETO: P53-Independent Upregulation of P21/WAF/CIP1.
Authors: Berg Tobias , Fliegauf Manfred , Pitako Jurij , Burger Jan , Abdelkarim Mahmoud , Guo Yalin , Staege Martin S. , Burdach Stefan , Lübbert Michael 
Published: April 2020

Abstract
Background: The translocation (8;21) is the most common chromosomal rearrangement in AML, resulting in the expression of the fusion protein AML1/ETO. We have developed an ecdysone-inducible U937 model, in which AML1/ETO is expressed in response to treatment with Ponasterone (Pon) A (Fliegauf et al, Oncogene 2004). This model system was used to determine the cellular effects of AML1/ETO and to identify its target genes in U937 cells.
Methods: Effects of AML1/ETO expression upon cell growth, viability, cell cycle and apoptosis were analyzed by trypan blue exclusion, FACS analysis using propidium iodide and DiOC6 staining, DNA laddering and Western blot for PARP cleavage, respectively. The gene expression profile of U937 with and without conditional AML1/ETO expression was assessed using Affymetrix U133A microarrays. Wild-type U937 cells with and without PonA treatment as well as AML1/ETO-negative and AML1/ETO-positive myeloid cell lines served as controls. Northern and Western Blotting were used for validation of expression changes.
Results: Induction of AML1/ETO expression in U937 resulted in reduced cell growth, G1 arrest and in apoptosis beginning 48–72 hours after PonA treatment. To investigate the underlying mechanisms, microarray analysis was performed. Expression profiles of AML1/ETO-positive and AML1/ETO-negative cell lines formed distinct clusters. Based on stringent criteria, 191 different genes were found upregulated, whereas 37 were downregulated upon expression of AML1/ETO in U937. The identified genes were screened for genes with known functions in cell cycle and apoptosis by automated and manual review and included 13 apoptosis-related genes. Among them, the CDK inhibitor p21/WAF/CIP1 was upregulated 19-fold upon induction of AML1/ETO, whereas the apoptosis regulator MCL-1 was induced 2.5-fold. Based on our criteria, no differential expression of other transcriptionally-controlled apoptosis regulators (such as BCL2, BAX, BAK1, BAD or c-flip) was noted. Northern and Western Blot analysis confirmed the strong induction of p21/WAF/CIP1 that paralleled the expression of AML1/ETO 10 hours after PonA treatment. Induction of p21/WAF/CIP1 was independent of the tumor suppressor protein p53 (Dou et al., Proc. Natl. Acad. Sci. 1995), and by Western blot, p53 was undetectable in U937. Northern Blot analysis revealed a higher expression of p21/WAF/CIP1 in the AML1/ETO-positive cell lines Kasumi-1 and SKNO-1 than in the AML1/ETO-negative cell lines HL-60, KG-1 and U937, supporting our finding that AML1/ETO may induce p21/WAF/CIP1.
Conclusions: AML1/ETO expression resulted in increased expression of p21/WAF/CIP1, which might contribute to the observed growth arrest and induction of apoptosis caused by the conditional expression of AML1/ETO.
High Rate of Deep-Vein Thrombosis in Asia after Major Orthopedic Surgery: The AIDA Study Final Results.
Authors: Piovella Franco , Wang Ching-Jen , Lu Housan , Kenneth Lee , Lee Lai Heng , Lee Won Chul , Turpie Alexander G. , Gallus Alexander , Planes Andre , Passera Raffaella , Rouillon Alain ,   The AIDA Investigators
Published: April 2020

Abstract
Background: The incidence of post-surgical venous thromboembolism is thought to be low in Asian populations and routine thromboprophylaxis is not implemented.
Objective: In this large multinational, multiethnic study, we evaluated the incidence of deep-vein thrombosis (DVT) patients undergoing major orthopedic surgery of the lower limbs.
Methods: We performed a prospective epidemiological study in 19 centers across Asia (China, Indonesia, South Korea, Malaysia, Philippines, Taiwan and Thailand) in patients undergoing elective total hip replacement (THR), total knee replacement (TKR) or hip fracture surgery (HFS) not receiving pharmacological thromboprophylaxis. The primary endpoint was the rate of DVT of the lower limbs objectively documented by mandatory bilateral ascending venography performed 6 to 10 days after surgery and evaluated by a blinded central adjudication committee.
Results: Overall, 407 Asian patients (20–99 years) undergoing THR (n=175), TKR (n=136) or HFS (n= 96) were recruited in 19 centres. 72.5% of the enrolled patients had adequate venograms. Total DVT was diagnosed in 121 of 295 evaluable patients (41.0%, [95% confidence interval: 35.4–46.7]). Proximal DVT was found in 30 patients (10.2% [7.0–14.2]). Total DVT and proximal DVT rates were the highest in TKR patients (58.1% and 17.1%, respectively), followed by HFS patients (42.0% and 7.2%, respectively), then THR patients (25.6% and 5.8%, respectively). DVT was more frequent in female patients aged at least 65 years. By-country analysis showed that the highest DVT rate was observed in Indonesia (78.6%) and the lowest in China (24.3%). By ethnic group, the lowest DVT rate was observed in Koreans (29.8%), while similar results were found in Chinese (45.0%) and in other non-Korean non-Chinese (46.3%) ethnic groups. Pulmonary embolism was clinically suspected in 10 of 407 patients (2.5%) and objectively confirmed in two patients (0.5%).
Conclusions: Venographically detected thrombosis is well recognized as an appropriate surrogate for symptomatic outcomes and is the basis for the recommendation for routine prophylaxis in Western patients. The high rate of venographic thrombosis in Asian patients is similar to that observed in patients in Western countries. Thrombosis prophylaxis should therefore be considered in Asian patients undergoing major orthopedic procedures.
Lipoprotein (A) and Further Prothrombotic Risk Factors in Women with Unexplained Recurrent Miscarriage- Results of a Multicenter Case-Control Study.
Authors: Nowak-Gottl Ulrike , Sonntag Barbara , Klamroth Robert , Heinecke Achim , Langer Claus , Scharrer Inge , Greb Robert , Krause Manuela 
Published: April 2020

Abstract
Background The present study was performed to assess the association of prothrombotic risk factors and underlying conditions, e.g. smoking and obesity, with unexplained recurrent miscarriage (uRM) in white women.
Methods and Results From 1998 to 2003, 133 Caucasian women aged 18–42 years (median 28 years) suffering from uRM were consecutively enrolled. In patients and 133 age-matched healthy controls prothrombotic risk factors (factor V (FV) G1691A, factor II (FII) G20210A, MTHFR T677T, 4G/5G plasminogen activator inhibitor (PAI)-1, lipoprotein (Lp) (a), protein C (PC), protein S (PS), antithrombin (AT), antiphospholipid/anticardiolipin (APA/ACA) antibodies) as well as associated environmental conditions were investigated. 70 (52.6%) of the patients had at least one prothrombotic risk factor compared with 26 control women (19.5%; p&lt;0.0001). Body mass index (BMI; p=0.78) and smoking habits (p=0.44) did not differ significantly between the groups investigated. Upon univariate analysis the heterozygous FV mutation, Lp(a) &gt; 30 mg/dL, increased APA/ACA and BMI &gt; 25 kg/m2 in combination with a prothrombotic risk factor were found to be significantly associated with uRM. Associations of the heterozygous FII variant, MTHFR or 4G/4G PAI-1 genotypes, deficiency states of PC, PS and AT, as well as the combination of smoking with at least one prothrombotic risk factor did not reach statistical significance. In multivariate analysis, increased Lp(a) (odds ratio (OR): 4.7/95% confidence interval (CI): 2.0–10.7), the FV mutation (OR:3.8/CI:1.4–10.7), and increased APA/ACA (OR: 4.5/CI: 1.1–17,7) had independent associations with uRM.
Conclusion In Caucasian women uRM is associated with the presence of elevated Lp(a) or further prothrombotic risk factors.
Comparison of Compression Ultrasound vs Ascending Contrast Venography for Proximal DVT in Medical-Surgical ICU Patients.
Authors: Crowther Mark A. , Bates Shannon , Schiff David , Dobranowski Julian , Meade Maureen , McDonald Ellen , Geerts Bill , Cook Deborah 
Published: April 2020

Abstract
Background: Ascending contrast venography is the reference standard for diagnosing DVT. Observational studies suggest that compression ultrasound has 97% sensitivity and 94% specificity for proximal DVT in symptomatic patients. However, in asymptomatic patients, such as those in the medical-surgical ICU, the test properties of ultrasound may be substantially worse. Surveys, observational studies and professional documents show that many intensivists, Research Ethics Boards, and the American College of Radiology have rejected the routine use of venography in the ICU setting.
Objective: To compare the results of positive lower limb compression ultrasound with ascending contrast venography in medical-surgical ICU patients.
Design: One year longitudinal cohort study.
Setting: 15 bed medical-surgical closed ICU in Hamilton, Canada.
Methods: We enrolled consecutive patients age &gt;18y with an expected ICU admission &gt;72h. Exclusion criteria were trauma, orthopedic surgery, cardiac surgery, pregnancy or palliative care. Patients underwent bedside lower limb compression ultrasonography by a certified ultrasonographer on ICU admission, twice weekly, and upon clinical suspicion of VTE. Patients with a positive or nondiagnostic ultrasound underwent bilateral lower extremity ascending contrast venography, unless they had pre-specified contraindications: serum creatinine &gt;150 umol/L, pre-existing insulin-requiring diabetes, known or suspected contrast allergy, or contraindications to transport to the Radiology Department. The study radiologist and thrombosis consultant independently interpreted the venograms in duplicate; disagreements were resolved by a third adjudicator.
Results: Of 261 patients, lower limb DVT was identified by compression ultrasound in 32 patients (7, 2.7% on ICU admission and 25, 9.6% during the ICU stay). Using the a priori venogram eligibility criteria, 8 of 32 patients (25.0%) with DVT by screening ultrasound were suitable for venography; 7 of the 8 venograms (87.5%) were positive.
Conclusions: Venogram eligibility criteria (e.g., low oxygen requirements, hemodynamic stability, and low risk for contrast nephropahthy) designed to safely confirm or refute positive or nondiagnostic screening ultrasound findings resulted in only selected, less seriously ill ICU patients undergoing venography. Concerns about the risk: benefit ratio of research interventions not used in current practice means that in multicenter trials, serial compression ultrasound will be the only safe and feasible method of screening for proximal DVT in the medical-surgical critically ill population.
Non-Genomic Effects of Estradiol on Human Platelets: Regulation of Mitogen Activated Protein Kinase Activation and Enhanced Spreading and Aggregation.
Authors: Vijayan Kadekuzhi V. , Liu Yan , Li Tong-Tong , Bray Paul F. 
Published: April 2020

Abstract
17β-estradiol or estrogen (E2) is a sex hormone that modulates platelet function and is widely used in hormone replacement therapy (HRT). We and others have previously demonstrated that human megakaryocytes and platelets posses estrogen receptors ERα and ERβ. HRT treatment augmented the bone marrow megakaryocytes without increasing other bone marrow cells, suggesting that E2 can modulate proliferation of megakaryocytes. Since mitogen activated protein kinases (MAPKs) are critical for 1) cell proliferation, 2) megakarytocyte differentiation and proplatelet formation and estrogen activates MAPK in other cell types, we hypothesized that estrogen regulates the activation of MAPK in human platelets. Signaling was studied using washed platelets from male and female subjects in response to varying concentrations of estrogen. Compared to the ethanol (vehicle) treated platelets, 1 nM E2 treated platelets for 60 seconds resulted in an enhanced activation of extracellular signal-regulated kinase 2 (ERK 2) and P38 but not Jun N-kinase (JNK). These results suggest that E2 can cause a non-genomic signaling in human platelets. The MEK inhibitors PD98059 and U0216 blocked the E2 effect, suggesting that the activation of ERK 2 was mediated through the upstream mitogen activated protein kinase kinase (MAPKK). Because E2 can modulate actin reorganization in other cell types and cell spreading is promoted by ERK 2 activation, we examined the effect of E2 on platelet spreading - a process not dependent on agonist stimulation. Compared to ethanol treated platelets, platelets preincubated with 100 nM E2 for 60 seconds and stained with rhodamine phallodine exhibited a ~60 % greater spreading at 5 and 15 minutes. This observation suggests that that E2 can cause rapid actin cytoskeletal reorganization in platelets. Since inhibition of ERK 2 activation blocks aggregation to low doses of thrombin and collagen, we examined a role for E2 in platelet aggregation. E2 alone did not induce platelet aggregation. However, E2 potentiated aggregation with low but not high doses of thrombin and collagen related peptide (CRP) (P=0.05 for 0.02 μg/ml thrombin and P&lt;0.001 for 0.2 μg/ml CRP). Our data demonstrates that E2 can activate MAPK through a non-genomic mechanism and this activation correlates with greater platelet functions like spreading and aggregation. Our findings support a mechanism whereby a consistent non-genomic enhancement of platelet signaling and reactivity by E2 may underlie the increased cardiovascular events observed in recent randomized clinical trials with HRT.
Evaluation of Baseline Body-Weight Dosing of Yttrium 90 (90Y) Ibritumomab Tiuxetan (Zevalin®) Radioimmunotherapy in Patients with Relapsed or Refractory B-Cell Non-Hodgkin’s Lymphoma (NHL).
Authors: Wiseman Gregory , Conti Peter , Vo Katie , Schilder Russell J. , Foster Paul , Gordon Leo I. , Emmanouilides Christos , Silverman Dan , Witzig Thomas E. , Molina Arturo 
Published: April 2020

Abstract
Background: As the first radiolabled monoclonal antibody approved for the treatment for the treatment of cancer, yttrium 90 (90Y) ibritumomab tiuxetan (Zevalin®) achieves both high response rates and durable remissions in patients with relapsed or refractory, low-grade, follicular, or transformed B-cell NHL. Because of the stability of 90Y ibritumomab tiuxetan and its favorable pharmacokinetic profile (ie, predictable urinary clearance and absence of dehalogenation), dosing is based on patient weight and platelet counts. Yttrium 90 ibritumomab tiuxetan is typically delivered at a dose of 0.4 mCi/kg (in patients with platelets &gt;150,000/mm3) or 0.3 mCi/kg (in patients with platelets ≤150,000/mm3) to a maximum recommended dose (MRD) of 32 mCi. However, patients &gt;80 kg with platelet counts exceeding 150,000/mm3 may receive a lower drug concentration (ie, &lt;0.4 mCi/kg 90Y) as a result of dose capping. We evaluated whether the 32 mCi dose cap influences the safety or efficacy of ibritumomab tiuxetan therapy in patients &gt;80 kg.
Methods: Efficacy and safety data were collected for patients from 3 registrational trials that received either 0.4 mCi/kg 90Y (patients ≤80 kg) or 32 mCi 90Y (patients &gt;80 kg).
Results: Clinical responses in 170 patients were evaluated. Patients ≤80 kg (n = 103) had a median weight of 70 kg (range, 45–80 kg) versus a median weight of 95 kg (range, 81–159) for patients &gt;80 kg (n = 67). Gender (41% M vs 73% M, respectively; P &lt;.01) was the only significantly different baseline characteristic between the ≤80 kg and &gt;80 kg groups. Similar efficacy and safety results were reported for both subsets of patients. Overall response rates of 79% and 70% were observed for patients ≤80 kg and &gt;80 kg, respectively (P =.27); and no significant differences were seen in complete response rates (28% vs 34%, respectively; P =.40). Median TTP (8.9 months vs 9.5 months; P =.53) and median DR (8.5 months vs 11.5 months; P =.34) were equivalent between the 2 weight-based groups. In addition, there were no statistically significant differences in safety measures including grade 3/4 nonhematologic adverse events, neutropenia, thrombocytopenia, or anemia.
Conclusions: As a consequence of dose capping at an MRD of 32 mCi, 39% of patients received a lower dose/kg of 90Y ibritumomab tiuxetan. However, despite the difference in dose administered on a unit of body weight basis, the efficacy and safety results were similar between patients ≤80 kg and &gt;80 kg. We conclude that the 32 mCi dose cap does not influence the efficacy or safety profile of 90Y ibritumomab tiuxetan for patients &gt;80 kg.
Upregulation of the Chemokine Receptor CCR4 in Cutaneous T-Cell Lymphoma and Nodal T-Cell Lymphoma: Implications for Targeted Therapy.
Authors: Jones Dan , Patel Kaushali , Herling Marco , Aguilar Richard J. , Ueda Ryuzo , Inagaki Hiroshi , Fayad Luis , Duvic Madeleine 
Published: April 2020

Abstract
The chemokine receptor CCR4 is expressed by a minor population of non-neoplastic T-cells, particularly Th2 helper T-cells, skin-homing T-cells and CD4+CD8− thymocytes. To assess the suitability of this receptor as a target for immunotherapy, we compared expression of CCR4 in a large series of T-cell malignancies with non-neoplastic T-cells by immunohistochemistry on formalin-fixed, paraffin-embedded material. Using tissue arrays and whole tissue sections, we assessed CCR4 levels in 233 biopsy samples from 182 patients with various T-cell tumors, particularly mycosis fungoides (MF, n = 54) and Sezary syndrome (SS, n = 15). We utilized the anti-CCR4 antibody KM2160 (Kyowa Hakko Kogyo, Tokyo, Japan), whose specificity has been established by immunoprecipitation and flow cytometry on CCR4-expressing transfectants and T-cell lines. Immunostaining was graded on a 4-tier scale (negative, focal, partial and uniform positivity) and compared with staining for markers of tumor progression including CD25, CD30 and the cytotoxic marker granzyme B and with a mediator of lymphocyte homing to skin, cutaneous lymphocyte antigen (CLA). Overall, CCR4 was expressed either partially or uniformly in 49% of T-cell tumors. Among MF/SS cases, partial or uniform CCR4 expression was found in 52% of cases, including in both early patch/plaque stage lesions and disseminated disease. CCR4 was upregulated upon large cell transformation in 39% of 23 MF/SS cases studied in sequential biopsies. There was an 80% concordance rate in CCR4 staining patterns in tumor cells in biopsies taken simultaneously from different anatomic sites. Upregulation of CCR4 upon large cell transformation was independent of other markers of progression, including CD25, CD30 and granzyme B but was significantly correlated with the degree of CLA expression. In contrast to the strong intensity of CCR4 staining in MF/SS cases, dimmer more variable expression was seen in skin homing T-cells in 94% of chronic dermatitis cases (n = 18). Among other T-cell tumors, CCR4 expression was common in AIL (70%), ALCL (53%) and PTCL unspecified (55%) and in the more mature subset of precursor-T lymphoblastic lymphoma/leukemia. CCR4-expressing T-cells were rare in 12 lymph node biopsies with reactive lymphadenitis. In conclusion, the chemokine receptor CCR4 is stably upregulated in cutaneous lymphoma and nodal T-cell lymphoma as compared to levels seen in non-neoplastic T-cells and thus represents a promising antigen for selective targeting of T-cell malignancies by immunotherapy.