Costunolide causes mitotic arrest and enhances radiosensitivity in human hepatocellular carcinoma cells

<p>Abstract</p> <p>Purpose</p> <p>This work aimed to investigate the effect of costunolide, a sesquiterpene lactone isolated from <it>Michelia compressa</it>, on cell cycle distribution and radiosensitivity of human hepatocellular carcinoma (HCC) cells.</p> <p>Methods</p> <p>The assessment used in this study included: cell viability assay, cell cycle analysis by DNA histogram, expression of phosphorylated histone H3 (Ser 10) by flow cytometer, mitotic index by Liu's stain and morphological observation, mitotic spindle alignment by immunofluorescence of alpha-tubulin, expression of cell cycle-related proteins by Western blotting, and radiation survival by clonogenic assay.</p> <p>Results</p> <p>Our results show that costunolide reduced the viability of HA22T/VGH cells. It caused a rapid G2/M arrest at 4 hours shown by DNA histogram. The increase in phosphorylated histone H3 (Ser 10)-positive cells and mitotic index indicates costunolide-treated cells are arrested at mitosis, not G2, phase. Immunofluorescence of alpha-tubulin for spindle formation further demonstrated these cells are halted at metaphase. Costunolide up-regulated the expression of phosphorylated Chk2 (Thr 68), phosphorylated Cdc25c (Ser 216), phosphorylated Cdk1 (Tyr 15) and cyclin B1 in HA22T/VGH cells. At optimal condition causing mitotic arrest, costunolide sensitized HA22T/VGH HCC cells to ionizing radiation with sensitizer enhancement ratio up to 1.9.</p> <p>Conclusions</p> <p>Costunolide could reduce the viability and arrest cell cycling at mitosis in hepatoma cells. Logical exploration of this mitosis-arresting activity for cancer therapeutics shows costunolide enhanced the killing effect of radiotherapy against human HCC cells.</p

MGMRES: A generalization of GMRES for solving large sparse nonsymmetric linear systems

This paper is concerned with the solution of the linear system Au = b, where A is a real square nonsingular matrix which is large, sparse and nonsymmetric. We consider the use of Krylov subspace methods. We first choose an initial approximation u{sup (0)} to the solution {bar u} = A{sup -1}b. The GMRES (Generalized Minimum Residual Algorithm for Solving Non Symmetric Linear Systems) method was developed by Saad and Schultz (1986) and used extensively for many years, for sparse systems. This paper considers a generalization of GMRES; it is similar to GMRES except that we let Z = A{sup T}Y, where Y is a nonsingular matrix which is symmetric but not necessarily SPD

In Situ Monitoring of Temperature inside Lithium-Ion Batteries by Flexible Micro Temperature Sensors

Lithium-ion secondary batteries are commonly used in electric vehicles, smart phones, personal digital assistants (PDA), notebooks and electric cars. These lithium-ion secondary batteries must charge and discharge rapidly, causing the interior temperature to rise quickly, raising a safety issue. Over-charging results in an unstable voltage and current, causing potential safety problems, such as thermal runaways and explosions. Thus, a micro flexible temperature sensor for the in in-situ monitoring of temperature inside a lithium-ion secondary battery must be developed. In this work, flexible micro temperature sensors were integrated into a lithium-ion secondary battery using the micro-electro-mechanical systems (MEMS) process for monitoring temperature in situ

Security of Deputy Signature

E-system, a new commerce model, is a new era for business direction. When a principal is absent (goes on an errand or on leave), a well-designed deputy system keeps the business operations working. In the network world, identity verification and any substitute for traditional signature can be done by digital signature [1]. Deputy signature guarantees the existence of deputy system in e-system. Current deputy mechanism addresses the verification of deputy signature. No research has been done on the prevention of the illegal use of deputy system when the principal returns and the deputy system is not in use. We propose a mechanism to solve the problem of illegal use of deputy system when the power of deputy system is not legally “ON.

Stimulation of Type I Collagen Transcription in Human Skin Fibroblasts by TGF-β: Involvement of Smad 3

Transforming growth factor-β (TGF-β) stimulates the transcription of the α2(I) procollagen gene (COL1A2). The intracellular mediators involved in this response remain poorly understood. In this study, we demonstrate that primary human skin fibroblasts express Smads, a novel family of signaling molecules, in vitro in the absence of TGF-β. The levels of Smad 7 mRNA was rapidly and transiently increased by TGF-β. Transient overexpression of Smad 3 and Smad 4, but not Smad 1 or Smad 2, caused trans-activation of a CAT reporter gene driven by a 772 bp segment of the human COL1A2 promoter containing putative TGF-β response elements. Smad stimulation of promoter activity was ligand independent, but was further enhanced by TGF-β. Overexpression of a phosphorylation-deficient Smad 3 mutant or wild-type Smad 7, which lacks the carboxy-terminal phosphorylation motif, specifically inhibited TGF-β-induced activation of COL1A2 promoter. A CAGACA sequence shown to be a functional Smad-binding element in the plasminogen activator inhibitor-1 gene promoter was found within the TGF-β-response region of the proximal COL1A2 promoter. Gel mobility shift assays showed protein phosphorylation-dependent binding activity in fibroblast nuclear extracts specific for this sequence; TGF-β treatment strongly stimulated the formation of this DNA-protein complex. Smad was identified as a component of the CAGACA-binding transcription complex in TGF-β-treated fibroblasts by antibody supershifting. These results demonstrate that (i) Smad 3 transmits TGF-β signals from the receptor to the COL1A2 promoter in human fibroblasts, and is likely to play an important role in stimulation of COL1A2 promoter activity elicited by TGF-β; (ii) in fibroblasts, Smads appear to function as inducible DNA-binding transcription factors; and (iii) Smad 7 may be involved in autocrine negative feedback in the regulation of COL1A2 promoter activity by TGF-β