Myoferlin Depletion in Breast Cancer Cells Promotes Mesenchymal to Epithelial Shape Change and Stalls Invasion

Myoferlin (MYOF) is a mammalian ferlin protein with homology to ancestral Fer-1, a nematode protein that regulates spermatic membrane fusion, which underlies the amoeboid-like movements of its sperm. Studies in muscle and endothelial cells have reported on the role of myoferlin in membrane repair, endocytosis, myoblast fusion, and the proper expression of various plasma membrane receptors. In this study, using an in vitro human breast cancer cell model, we demonstrate that myoferlin is abundantly expressed in invasive breast tumor cells. Depletion of MYOF using lentiviral-driven shRNA expression revealed that MDA-MB-231 cells reverted to an epithelial morphology, suggesting at least some features of mesenchymal to epithelial transition (MET). These observations were confirmed by the down-regulation of some mesenchymal cell markers (e.g., fibronectin and vimentin) and coordinate up-regulation of the E-cadherin epithelial marker. Cell invasion assays using Boyden chambers showed that loss of MYOF led to a significant diminution in invasion through Matrigel or type I collagen, while cell migration was unaffected. PCR array and screening of serum-free culture supernatants from shRNAMYOF transduced MDA-MB-231 cells indicated a significant reduction in the steady-state levels of several matrix metalloproteinases. These data when considered in toto suggest a novel role of MYOF in breast tumor cell invasion and a potential reversion to an epithelial phenotype upon loss of MYOF

INFLUENCE OF SWEPT WING ON AXIAL STEAM TURBINE STATIC CASCADE AERODYNAMICS

ABSTRACT Super-critical steam turbine high pressure stage static blade cascade is as the prototype blade to numerical simulations. The prototype blade has aft-loading and big front-edge radius characteristics. Different swept blades are made by changing the sweep angle and sweep height in order to study the effect of blade sweep on aerodynamic characteristics of super-critical steam turbine high pressure stage blade. The numerical simulation sweep angle are made of fore-angel 10°, 20°, 30°a nd aft-angle 10°, 20°, 30°, sweep height are made of 10%, 20%, 30% blade height. The steam turbine aerodynamics are analyzed such as static pressure coefficient of swept blade profiles, total pressure coefficient of swept blade at blade trailing edge, contour of mass averaged static pressure on meridional surface and velocity streamlines along hub and suction surface by CFD software CFX®. The numerical simulation results indicate the fore-swept blades increase the low energy fluid centralizing in endwall corner and the endwall secondary flow loss, but the loss is decreased at mid-span, comparing to the baseline. But aft-swept blades decrease the low energy fluid centralizing in endwall corner and the endwall secondary flow loss, but the loss is increased at mid-span, comparing to the baseline. Either the fore-sweep height or the aft-sweep angle can increase the above results. At the same time the swept blade aerodynamics also are computed at 10,-10 degree incidence, the results indicate it keep down the aftloading characteristics of baseline

Numerical Analysis of Conjugated Heat Transfer and Thermal Stress Distributions in a High-Temperature Ni-Based Superalloy Turbine Rotor Blade

This paper establishes a multidisciplinary method combining conjugate heat transfer (CHT) and thermal stress for a high-temperature Ni-based superalloy turbine rotor blade with integrated cooling structures. A conjugate calculation is performed to investigate the coolant flow characteristics, heat transfer, and thermal stress of the rotor blade under rotating and stationary conditions to understand the effects of rotation on the multidisciplinary design of the blade. Furthermore, the maximum resolved shear stress among the 30-slip systems and the corresponding dominant slip system are obtained to predict the deformation tendency of the blade by employing the crystal plasticity finite element method (CPFEM) and considering the specified anisotropic blade material (GTD-111). The results show that the forces of rotation, including centrifugal and Coriolis forces, and their induced buoyancy force, alter the coolant flow field and thus affect the rotor blade’s heat transfer distribution compared with the stationary condition. The maximum temperature and thermal stress of the rotor blade under rotating conditions are reduced by 5% and 21% compared with that under the stationary condition, respectively. Compared with the stationary condition, the temperature and thermal stress distribution on the blade under the rotating condition are more uniform, especially on the suction side. In addition, the blade root connecting with the hub, the film holes near the leading-edge region at the blade root, the mid-chord of the suction surface, and the grooved blade tip are easily damaged by the enormous resolved shear stress and the interface effect of different types of dominant slip system under the two conditions. In this work, it was feasible to use the cascade cooling effect test to analyze the dynamic test results for the rotor blade. Furthermore, the thermal stress analysis based on the CPFEM can provide a superior level of blade cooling design than CHT by considering the anisotropic material characteristics of a turbine blade

Numerical Analysis of Conjugated Heat Transfer and Thermal Stress Distributions in a High-Temperature Ni-Based Superalloy Turbine Rotor Blade

This paper establishes a multidisciplinary method combining conjugate heat transfer (CHT) and thermal stress for a high-temperature Ni-based superalloy turbine rotor blade with integrated cooling structures. A conjugate calculation is performed to investigate the coolant flow characteristics, heat transfer, and thermal stress of the rotor blade under rotating and stationary conditions to understand the effects of rotation on the multidisciplinary design of the blade. Furthermore, the maximum resolved shear stress among the 30-slip systems and the corresponding dominant slip system are obtained to predict the deformation tendency of the blade by employing the crystal plasticity finite element method (CPFEM) and considering the specified anisotropic blade material (GTD-111). The results show that the forces of rotation, including centrifugal and Coriolis forces, and their induced buoyancy force, alter the coolant flow field and thus affect the rotor blade’s heat transfer distribution compared with the stationary condition. The maximum temperature and thermal stress of the rotor blade under rotating conditions are reduced by 5% and 21% compared with that under the stationary condition, respectively. Compared with the stationary condition, the temperature and thermal stress distribution on the blade under the rotating condition are more uniform, especially on the suction side. In addition, the blade root connecting with the hub, the film holes near the leading-edge region at the blade root, the mid-chord of the suction surface, and the grooved blade tip are easily damaged by the enormous resolved shear stress and the interface effect of different types of dominant slip system under the two conditions. In this work, it was feasible to use the cascade cooling effect test to analyze the dynamic test results for the rotor blade. Furthermore, the thermal stress analysis based on the CPFEM can provide a superior level of blade cooling design than CHT by considering the anisotropic material characteristics of a turbine blade

Increased biogenesis of glucagon-containing secretory granules and glucagon secretion in BIG3-knockout mice

OBJECTIVE: Although both insulin and glucagon are intimately involved in the regulation of glucose homeostasis, the intrinsic control of glucagon secretion, including the biogenesis and exocytosis of glucagon-containing granules, is far less understood compared with that of insulin. As Brefeldin A-inhibited guanine nucleotide exchange protein 3 (BIG3) is a negative regulator of insulin-granule biogenesis and insulin secretion, we investigated whether BIG3 plays any role in alpha-cells and glucagon secretion. METHODS: We examined the expression of BIG3 in islet cells by immuno-fluorescence and confocal microscopy, and measured glucagon production and secretion in BIG3-depleted and wild-type mice, islets and cells. RESULTS: BIG3 is highly expressed in pancreatic alpha-cells in addition to beta-cells, but is absent in delta-cells. Depletion of BIG3 in alpha-cells leads to elevated glucagon production and secretion. Consistently, BIG3-knockout (BKO) mice display increased glucagon release under hypoglycemic conditions. CONCLUSIONS: Together with our previous studies, the current data reveal a conserved role for BIG3 in regulating alpha- and beta-cell functions. We propose that BIG3 negatively regulates hormone production at the secretory granule biogenesis stage and that such regulatory mechanism may be used in secretory pathways of other endocrine cells.status: publishe

Increased biogenesis of glucagon-containing secretory granules and glucagon secretion in BIG3-knockout mice

Objective: Although both insulin and glucagon are intimately involved in the regulation of glucose homeostasis, the intrinsic control of glucagon secretion, including the biogenesis and exocytosis of glucagon-containing granules, is far less understood compared with that of insulin. As Brefeldin A-inhibited guanine nucleotide exchange protein 3 (BIG3) is a negative regulator of insulin-granule biogenesis and insulin secretion, we investigated whether BIG3 plays any role in alpha-cells and glucagon secretion. Methods: We examined the expression of BIG3 in islet cells by immuno-fluorescence and confocal microscopy, and measured glucagon production and secretion in BIG3-depleted and wild-type mice, islets and cells. Results: BIG3 is highly expressed in pancreatic alpha-cells in addition to beta-cells, but is absent in delta-cells. Depletion of BIG3 in alpha-cells leads to elevated glucagon production and secretion. Consistently, BIG3-knockout (BKO) mice display increased glucagon release under hypoglycemic conditions. Conclusions: Together with our previous studies, the current data reveal a conserved role for BIG3 in regulating alpha- and beta-cell functions. We propose that BIG3 negatively regulates hormone production at the secretory granule biogenesis stage and that such regulatory mechanism may be used in secretory pathways of other endocrine cells