Ligand-Induced Tyrosine Phosphorylation of Cysteinyl Leukotriene Receptor 1 Triggers Internalization and Signaling in Intestinal Epithelial Cells

Leukotriene D(4) (LTD(4)) belongs to the bioactive lipid group known as eicosanoids and has implications in pathological processes such as inflammation and cancer. Leukotriene D(4) exerts its effects mainly through two different G-protein-coupled receptors, CysLT(1) and CysLT(2). The high affinity LTD(4) receptor CysLT(1)R exhibits tumor-promoting properties by triggering cell proliferation, survival, and migration in intestinal epithelial cells. In addition, increased expression and nuclear localization of CysLT(1)R correlates with a poorer prognosis for patients with colon cancer

Mahanine restores RASSF1A expression by down-regulating DNMT1 and DNMT3B in prostate cancer cells

BACKGROUND: Hypermethylation of the promoter of the tumor suppressor gene RASSF1A silences its expression and has been found to be associated with advanced grade prostatic tumors. The DNA methyltransferase (DNMT) family of enzymes are known to be involved in the epigenetic silencing of gene expression, including RASSF1A, and are often overexpressed in prostate cancer. The present study demonstrates how mahanine, a plant-derived carbazole alkaloid, restores RASSF1A expression by down-regulating specific members of the DNMT family of proteins in prostate cancer cells. RESULTS: Using methylation-specific PCR we establish that mahanine restores the expression of RASSF1A by inducing the demethylation of its promoter in prostate cancer cells. Furthermore, we show that mahanine treatment induces the degradation of DNMT1 and DNMT3B, but not DNMT3A, via the ubiquitin-proteasome pathway; an effect which is rescued in the presence of a proteasome inhibitor, MG132. The inactivation of Akt by wortmannin, a PI3K inhibitor, results in a similar down-regulation in the levels DNMT1 and DNMT3B. Mahanine treatment results in a decline in phospho-Akt levels and a disruption in the interaction of Akt with DNMT1 and DNMT3B. Conversely, the exogenous expression of constitutively active Akt inhibits the ability of mahanine to down-regulate these DNMTs, suggesting that the degradation of DNMT1 and DNMT3B by mahanine occurs via Akt inactivation. CONCLUSIONS: Taken together, we show that mahanine treatment induces the proteasomal degradation of DNMT1 and DNMT3B via the inactivation of Akt, which facilitates the demethylation of the RASSF1A promoter and restores its expression in prostate cancer cells. Therefore, mahanine could be a potential therapeutic agent for advanced prostate cancer in men when RASSF1A expression is silenced

Pressure shielding mechanism of canopies for trailing edge noise reduction in aerofoils

Session: Airframe / High-Lift Noise IView Video Presentation: https://doi.org/10.2514/6.2023-3204.vidThe pressure shielding mechanism of bio-inspired surface treatment, called canopies, has been investigated experimentally and applied to reduce trailing edge noise generated by aero- foils. Surface pressure experiments beneath the boundary layer on a flat and aerofoil section show that canopies can attenuate surface pressure in two frequency ranges, ∆ f 1 = 0.1 to 1.0 kHz and ∆f2 = 2 to 12 kHz, at some critical canopies’ height from the wall. Canopies with an Open-Area-Ratio (OAR or σ) of 50 % placed closer (h/δ=0.08) to the surface tend to in- crease attenuation with frequency, without any low-frequency peak attenuation. This high- frequency attenuation is mainly due to the mechanism of dissipation, of small-scale structures in the boundary-layer, provided by the canopies, which have relatively higher wall shear stress compared to flat plate or thicker canopy designs. As h/δ increases, the low-frequency atten- uation in the surface pressure becomes noticeable, with a peak value of 5 dB for a critical height of h/δ∗ ∼ 1, indicating the mechanism of blockage or shielding of large structures in the boundary-layer is responsible for the low-frequency attenuation. For h/δ ≥ 0.16, both the low- and high-frequency attenuation reduces and becomes almost zero for h/δ = 0.5. Furthermore, the mechanism of pressure shielding provided by the canopy treatment is shown to be a local phenomenon, for 70% <OAR < 90 % and very sensitive to the location of the canopy itself. The maximum attenuation in surface pressure is seen for the canopy geometries with small rod diameters with less spacing. The optimum canopy geometry, based on the surface pres- sure studies, was applied near the trailing edge of the NACA0012 aerofoil. The far-field noise study demonstrates, for the first time, that canopies can reduce broadband noise levels up to 12-14 dB in the frequency range between 2 and 12 kHz, provided they are scaled appropriately based on the incoming turbulent boundary-layer flow.EPSRC under grant No. EP/V038273/1

Modified TNO-Blake model for aerofoil surface pressure prediction with canopies

Session: Airframe / High-Lift Noise IView Video Presentation: https://doi.org/10.2514/6.2023-3203.vidThe modelling of the surface pressure spectrum beneath a turbulent boundary layer near the trailing edge of an aerofoil with bio-inspired surface treatments, called canopies, is investigated. Canopies are simply a cylindrical rods uniformly spaced along the span of the aerofoil. The velocity measurements indicated that the flow at the trailing edge of an aerofoil treated with canopies is localised and shows periodic behaviour across the span with treatment spacing. As a result, the mean-flow velocity gradient along the span (@U1=@x3) cannot be assumed as zero for x2=h = 4, which is shown in this paper. Therefore, the original surface pressure solution of Poisson’s equation is modified by introducing an additional source term consisting of the mean-shear contribution, given as @U1=@x3 @u3=@x1. Furthermore, the surface pressure attenuation due to the canopies shows a periodic behaviour across the span for treatments with an Open-Area-Ratio (OAR) between 70% and 90 %. This observation is consistent with our previous experimental results; therefore, the primary motivation for proposing a 3D TNO-Blake model, accounting for the interaction between the gradient of the stream-wise mean velocity along the span and span-wise fluctuating component along the stream. The model is built based on the inputs from Large Eddy Simulation results and additional wind tunnel measurements.EPSRC under grant No EP/V038273/1

β-Catenin is involved in alterations in mitochondrial activity in non-transformed intestinal epithelial and colon cancer cells

BACKGROUND: Alteration in respiratory activity and mitochondrial DNA (mtDNA) transcription seems to be an important feature of cancer cells. Leukotriene D(4) (LTD(4)) is a proinflammatory mediator implicated in the pathology of chronic inflammation and cancer. We have shown earlier that LTD(4) causes translocation of beta-catenin both to the mitochondria, in which it associates with the survival protein Bcl-2 identifying a novel role for beta-catenin in cell survival, and to the nucleus in which it activates the TCF/LEF transcription machinery. METHODS: Here we have used non-transformed intestinal epithelial Int 407 cells and Caco-2 colon cancer cells, transfected or not with wild type and mutated (S33Y) beta-catenin to analyse its effect on mitochondria activity. We have measured the ATP/ADP ratio, and transcription of the mtDNA genes ND2, ND6 and 16 s in these cells stimulated or not with LTD(4). RESULTS: We have shown for the first time that LTD(4) triggers a cellular increase in NADPH dehydrogenase activity and ATP/ADP ratio. In addition, LTD(4) significantly increased the transcription of mtDNA genes. Overexpression of wild-type beta-catenin or a constitutively active beta-catenin mutant mimicked the effect of LTD(4) on ATP/ADP ratio and mtDNA transcription. These elevations in mitochondrial activity resulted in increased reactive oxygen species levels and subsequent activations of the p65 subunit of NF-kappaB. CONCLUSIONS: The present novel data show that LTD(4), presumably through beta-catenin accumulation in the mitochondria, affects mitochondrial activity, lending further credence to the idea that inflammatory signalling pathways are intrinsically linked with potential oncogenic signals

LPA5 Is Abundantly Expressed by Human Mast Cells and Important for Lysophosphatidic Acid Induced MIP-1β Release

Background: Lysophosphatidic acid (LPA) is a bioactive lipid inducing proliferation, differentiation as well as cytokine release by mast cells through G-protein coupled receptors. Recently GPR92/LPA5 was identified as an LPA receptor highly expressed by cells of the immune system, which prompted us to investigate its presence and influence on mast cells. Principal Findings: Transcript analysis using quantitative real-time PCR revealed that LPA5 is the most prevalent LPA-receptor in human mast cells. Reduction of LPA5 levels using shRNA reduced calcium flux and abolished MIP-1β release in response to LPA. Conclusions: LPA5 is a bona fide LPA receptor on human mast cells responsible for the majority of LPA induced MIP-1β release

Distinct Effects of Unfractionated Heparin versus Bivalirudin on Circulating Angiogenic Peptides

Background: Human studies of therapeutic angiogenesis, stem-cell, and progenitor-cell therapy have failed to demonstrate consistent clinical benefit. Recent studies have shown that heparin increases circulating levels of anti-angiogenic peptides. Given the widely prevalent use of heparin in percutaneous and surgical procedures including those performed as part of studies examining the benefit of therapeutic angiogenesis and cell-based therapy, we compared the effects of unfractionated heparin (UFH) on angiogenic peptides with those of bivalirudin, a relatively newer anticoagulant whose effects on angiogenic peptides have not been studied. Methodology/Principal Findings: We measured soluble fms-like tyrosine kinase-1 (sFLT1), placental growth factor (PlGF), vascular endothelial growth factor (VEGF), and soluble Endoglin (sEng) serum levels by enzyme linked immunosorbent assays (ELISA) in 16 patients undergoing elective percutaneous coronary intervention. Compared to baseline values, sFLT1 and PlGF levels increased by 26296313 % and 253654%, respectively, within 30 minutes of UFH therapy (p,0.01 for both; n = 8). VEGF levels decreased by 93.265 % in patients treated with UFH (p,0.01 versus baseline). No change in sEng levels were observed after UFH therapy. No changes in sFLT1, PlGF, VEGF, or sEng levels were observed in any patients receiving bivalirudin (n = 8). To further explore the direct effect of anticoagulation on circulating angiogenic peptides, adult, male wild-type mice received venous injections of clinically dosed UFH or bivalirudin. Compared to saline controls, sFLT1 an