Phosphorylation of RNA helicase A by DNA-dependent protein kinase is indispensable for expression of the MDR1 gene product P-glycoprotein in multidrug-resistant human leukemia cells

Development of multidrug resistance (MDR) in cancer frequently involves overexpression of the MDR1 gene product P-glycoprotein (P-gp), a drug transporter which severely impedes the efficacy of chemotherapy. Because intensive efforts to identify therapeutics that reverse MDR by inhibiting the drug transport activity of P-gp have not yet met with success, we have focused on the alternative strategy of targeting MDR1 promoter activation to knockdown P-gp expression in cancer cells. We recently identified RNA helicase A (RHA) inhibition as a rational strategy to downregulate P-gp in leukemia cells by showing that RHA RNAi knockdown abrogated P-gp expression in MDR variants of human leukemia HL-60 cells. In that report, we also demonstrated that RHA activated the MDR1 promoter in the MDR variant cells but not in the drug-sensitive counterpart. This led us to hypothesize that P-gp induction by RHA required cooperation with another factor present only in the MDR variants. Here, we identify the RHA cooperating factor as DNA-PK catalytic subunit (cs), and we show that DNA-PKcs resides with RHA at the MDR1 promoter in a multiprotein complex. Furthermore, targeted DNA-PKcs inhibition abrogated P-gp expression in the MDR variant cells. We demonstrate that constitutive multisite RHA phosphorylation producing retarded migration in SDS-PAGE is catalyzed by DNA-PKcs in the MDR variants, and does not occur in the parental cells, which are DNA-PKcs deficient. The indispensable role played by DNA-PK in P-gp overexpression in MDR leukemia cells in this report identifies targeted DNA-PK inhibition as a rational strategy to reverse drug resistance in cancer.     

Risk Factors for Blood Transfusions in Primary Anatomic and Reverse Total Shoulder Arthroplasty for Osteoarthritis

BackgroundThe purpose of this study was to determine risk factors for blood transfusion in primary anatomic and reverse total shoulder arthroplasty (TSA) performed for osteoarthritis.MethodsPatients who underwent anatomic or reverse TSA for a diagnosis of primary osteoarthritis were identified in a national surgical database from 2005 to 2018 by utilizing both CPT and ICD-9/ICD-10 codes. Univariate analysis was performed on the two transfused versus non-transfused cohorts to compare for differences in comorbidities and demographics. Independent risk factors for perioperative blood transfusions were identified via multivariate regression models.Results305 transfused and 18,124 nontransfused patients were identified. Female sex (p<0.001), age >85 years (p=0.001), insulin-dependent diabetes mellitus (p=0.001), dialysis dependence (p=0.001), acute renal failure (p=0.012), hematologic disorders (p=0.010), disseminated cancer (p<0.001), ASA ≥ 3 (p<0.001), and functional dependence (p=0.001) were shown to be independent risk factors for blood transfusions on multivariate logistic regression analysis.ConclusionSeveral independent risk factors for blood transfusion following anatomic/reverse TSA for osteoarthritis were identified. Awareness of these risk factors can help surgeons and perioperative care teams to both identify and optimize high-risk patients to decrease both transfusion requirements and its associated complications in this patient population. Level of Evidence: III     

Distinct Phenotypic Differences Associated with Differential Amplification of Receptor Tyrosine Kinase Genes at 4q12 in Glioblastoma

Gene amplification at chromosome 4q12 is a common alteration in human high grade gliomas including glioblastoma, a CNS tumour with consistently poor prognosis. This locus harbours the known oncogenes encoding the receptor tyrosine kinases PDGFRA, KIT, and VEGFR2. These receptors are potential targets for novel therapeutic intervention in these diseases, with expression noted in tumour cells and/or associated vasculature. Despite this, a detailed assessment of their relative contributions to different high grade glioma histologies and the underlying heterogeneity within glioblastoma has been lacking. We studied 342 primary high grade gliomas for individual gene amplification using specific FISH probes, as well as receptor expression in the tumour and endothelial cells by immunohistochemistry, and correlated our findings with specific tumour cell morphological types and patterns of vasculature. We identified amplicons which encompassed PDGFRA only, PDGFRA/KIT, and PDGFRA/KIT/VEGFR2, with distinct phenotypic correlates. Within glioblastoma specimens, PDGFRA amplification alone was linked to oligodendroglial, small cell and sarcomatous tumour cell morphologies, and rare MGMT promoter methylation. A younger age at diagnosis and better clinical outcome in glioblastoma patients is only seen when PDGFRA and KIT are co-amplified. IDH1 mutation was only found when all three genes are amplified; this is a subgroup which also harbours extensive MGMT promoter methylation. Whilst PDGFRA amplification was tightly linked to tumour expression of the receptor, this was not the case for KIT or VEGFR2. Thus we have identified differential patterns of gene amplification and expression of RTKs at the 4q12 locus to be associated with specific phenotypes which may reflect their distinct underlying mechanisms.     

Chemical composition and biological activities of essential oils of Salvia officinalis aerial parts as affected by diurnal variations

The main objective of this work is to evaluate the impact of the diurnal variation on the essential oil (EO) of Salvia officinalis and on their antioxidant, antifungal and insecticidal potentials. Obtained results showed that the chemical composition of EOs of sage varied significantly during the day. For the EO, the 7 am extract was characterized by the most significant antiradical activity. The EOs of 12 and 5 pm used at a dose of 10 μL were found to have the most effective potential to inhibit the growth of Botrytis cinerea whereas, the EO of 5 pm used at the same dose (10 μL) was the most effective against Fusarium sambucinum. For the fumigant test, the EO from 7 am had the highest activity against Spodoptera littoralis. The EO of 12 pm had the largest repellency activity against Trogoderma granarium. In addition, the EO from 7 am belongs to the repulsive class III, those of 12 and 5 pm belong to the repulsive class IV. The results of this study indicate how to optimize the best harvesting hour to obtain extracts characterized by the best yield of active compounds and by the more effective biological activity.     

The rs9939609 polymorphism in the fat mass and obesity associated (FTO) gene is associated with obesity in Tunisian population

Abstract

Context: Variations in the fat mass and obesity-associated gene (FTO) has been associated with obesity in many populations, but the results are conflicting.

Objective: The aim of this study was to evaluate the effect of the rs9939609 polymorphism in the FTO gene on obesity risk and plasma leptin, adiponectin, insulin and lipid concentrations in Tunisians.

Materials and methods: Four hundred and ninety-four subjects with obesity and 334 non-obese participated in this study. The rs9939609 (T/A) genotype was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.

Results: Significant differences in genotype frequencies were observed between cases and controls. In the separate analysis by gender, the association between the AA genotype and obesity was statistically significant in women but not in men. After stratification by obesity class this association remains only with obesity class III.

Discussion: Our study is in agreement with studies on Caucasian, Portuguese and Cebu Filipino populations where a gender-specific association was found between rs9939609 polymorphism and obesity. It is also in agreement with studies on Mexican, Spanish and European populations, where an association was found with obesity class III.

Conclusion: The rs9939609 polymorphism of FTO gene is associated with obesity, especially obesity class III in women.