Inhibition of cell adhesion by phosphorylated Ezrin/Radixin/Moesin

Altered phosphorylation status of the C-terminal Thr residues of Ezrin/Radixin/Moesin (ERM) is often linked to cell shape change. To determine the role of phophorylated ERM, we modified phosphorylation status of ERM and investigated changes in cell adhesion and morphology. Treatment with Calyculin-A (Cal-A), a protein phosphatase inhibitor, dramatically augmented phosphorylated ERM (phospho-ERM). Cal-A-treatment or expression of phospho-mimetic Moesin mutant (Moesin-TD) induced cell rounding in adherent cells. Moreover, reattachment of detached cells to substrate was inhibited by either treatment. Phospho-ERM, Moesin-TD and actin cytoskeleton were observed at the plasma membrane of such round cells. Augmented cell surface rigidity was also observed in both cases.

Meanwhile, non-adherent KG-1 cells were rather rich in phospho-ERM. Treatment with Staurosporine, a protein kinase inhibitor that dephosphorylates phospho-ERM, up-regulated the integrin-dependent adhesion of KG-1 cells to substrate.

These findings strongly suggest the followings: (1) Phospho-ERM inhibit cell adhesion, and therefore, dephosphorylation of ERM proteins is essential for cell adhesion. (2) Phospho-ERM induce formation and/or maintenance of spherical cell shape. (3) ERM are constitutively both phosphorylated and dephosphorylated in cultured adherent and non-adherent cells.     

Phosphatidylethanolamine Deficiency Impairs Escherichia coli Adhesion by Downregulating Lipopolysaccharide Synthesis,Which is Reversible by High Galactose/Lactose Cultivation

As the initiation step of bacterial infection or biofouling, bacterial adhesion on cells or substrates is generally an optimal target for antibacterial design. Phosphatidylethanolamine (PE) is the principal phospholipid in bacteria, and its function in bacterial adhesion remains unclear. In this study, four E. coli strains including two PE-deficient mutants (PE^?PC^? and PE^?PC^+?strains) and two PE-containing wild-type controls (PE?⁺?PC^? strains) were recruited to investigate the influence of PE deficiency on bacterial adhesion. We found that PE deficiency could impair E. coli adhesion on macrophages (human THP-1-derived and mouse RAW264.7 macrophages) or glass coverslips by downregulating lipopolysaccharide (LPS) biosynthesis, which could be reversible by high galactose/lactose but not glucose cultivation. The data imply that PE play important role in bacterial adhesion probably via affecting LPS biosynthesis and suggest that targeting PE biosynthesis is also a potential antibacterial strategy.     

Systemic sclerosis sera affect fibrillin-1 deposition by dermal blood microvascular endothelial cells: therapeutic implications of cyclophosphamide

Introduction

Systemic sclerosis (SSc) is a connective tissue disorder characterized by endothelial cell injury, autoimmunity and fibrosis. The following three fibrillin-1 alterations have been reported in SSc. (1) Fibrillin-1 microfibrils are disorganized in SSc dermis. (2) Fibrillin-1 microfibrils produced by SSc fibroblasts are unstable. (3) Mutations in the FBN1 gene and anti-fibrillin-1 autoantibodies have been reported in SSc. Fibrillin-1 microfibrils, which are abundantly produced by blood and lymphatic microvascular endothelial cells (B-MVECs and Ly-MVECs, respectively), sequester in the extracellular matrix the latent form of the potent profibrotic cytokine transforming growth factor β (TGF-β). In the present study, we evaluated the effects of SSc sera on the deposition of fibrillin-1 and microfibril-associated glycoprotein 1 (MAGP-1) and the expression of focal adhesion molecules by dermal B-MVECs and Ly-MVECs.

Methods

Dermal B-MVECs and Ly-MVECs were challenged with sera from SSc patients who were treatment-naïve or under cyclophosphamide (CYC) treatment and with sera from healthy controls. Fibrillin-1/MAGP-1 synthesis and deposition and the expression of α_vβ₃ integrin/phosphorylated focal adhesion kinase and vinculin/actin were evaluated by immunofluorescence and quantified by morphometric analysis.

Results

Fibrillin-1 and MAGP-1 colocalized in all experimental conditions, forming a honeycomb pattern in B-MVECs and a dense mesh of short segments in Ly-MVECs. In B-MVECs, fibrillin-1/MAGP-1 production and α_vβ₃ integrin expression significantly decreased upon challenge with sera from naïve SSc patients compared with healthy controls. Upon challenge of B-MVECs with sera from CYC-treated SSc patients, fibrillin-1/MAGP-1 and α_vβ₃ integrin levels were comparable to those of cells treated with healthy sera. Ly-MVECs challenged with SSc sera did not differ from those treated with healthy control sera in the expression of any of the molecules assayed.

Conclusions

Because of the critical role of fibrillin-1 in sequestering the latent form of TGF-β in the extracellular matrix, its decreased deposition by B-MVECs challenged with SSc sera might contribute to dermal fibrosis. In SSc, CYC treatment might limit fibrosis through the maintenance of physiologic fibrillin-1 synthesis and deposition by B-MVECs.     

Synthesis,biological evaluation,and molecular docking studies of novel 2-styryl-5-nitroimidazole derivatives containing 1,4-benzodioxan moiety as FAK inhibitors with anticancer activity

A series of 2-styryl-5-nitroimidazole derivatives containing 1,4-benzodioxan moiety (3a–3r) has been designed, synthesized and their biological activities were also evaluated as potential antiproliferation and focal adhesion kinase (FAK) inhibitors. Among all the compounds, 3p showed the most potent activity in vitro which inhibited the growth of A549 with IC₅₀ value of 3.11 μM and Hela with IC₅₀ value of 2.54 μM respectively. Compound 3p also exhibited significant FAK inhibitory activity (IC₅₀ = 0.45 μM). Docking simulation was performed for compound 3p into the FAK structure active site to determine the probable binding model.     

Retention of bacteria on a substratum surface with micro-patterned hydrophobicity

Bacteria adhere to almost any surface, despite continuing arguments about the importance of physico-chemical properties of substratum surfaces, such as hydrophobicity and charge in biofilm formation. Nevertheless, in vivo biofilm formation on teeth and also on voice prostheses in laryngectomized patients is less on hydrophobic than on hydrophilic surfaces. With the aid of micro-patterned surfaces consisting of 10-microm wide hydrophobic lines separated by 20-microm wide hydrophilic spacings, we demonstrate here, for the first time in one and the same experiment, that bacteria do not have a strong preference for adhesion to hydrophobic or hydrophilic surfaces. Upon challenging the adhering bacteria, after deposition in a parallel plate flow chamber, with a high detachment force, however, bacteria were easily wiped-off hydrophobic lines, most notably when these lines were oriented parallel to the direction of flow. Adhering bacteria detached slightly less from the hydrophilic spacings in between, but preferentially accumulated adhering on the hydrophilic regions close to the interface between the hydrophilic spacings and hydrophobic lines. It is concluded that substratum hydrophobicity is a major determinant of bacterial retention while it hardly influences bacterial adhesion.     

C2H2＋Ar处理医用涤纶材料的细菌粘附

目的：对最常用心脏血管替代材料涤纶片作最新发展的具有全方位表面改性特征的混合等离子体浸没注入,以观察经处理后的涤纶片抑制细菌粘附的效果。方法：用多功能全方位等离子体浸没及离子注入机(PⅢ),用射频电源建立气体等离子体,对涤纶材料作全方位乙炔和氩气混合离子(C2H Ar)注入获取表面改性涤纶片。用金黄色葡萄球菌,表皮葡萄球菌,大肠杆菌,绿脓杆菌,白色念珠菌制取细菌悬液并作5-^125I-2’-脱氧尿嘧啶核苷(^125I-UDR)标记,再对改性涤纶材料作体外细菌动态粘附实验。结果：表面改性涤纶材料改变了亲水性和表面能,降低了水分子接触角。与未改性材料相比,改性涤纶材料抗细菌粘附能力有较明显提高。结论：混合离子(C2H2 Ar)表面改性涤纶片有良好的抗细菌和血小板粘附能力。     

Ti2448合金种植体表面不同纳米管径生物活性膜对成骨细胞早期黏附的影响

观察Ti2448合金表面不同纳米管径生物活性膜对成骨细胞早期黏附的影响,筛选出能够抵抗细菌在种植体表面黏附和定植的最适管径,为研制开发具有抗菌性能的种植体提供实验基础。采用阳极氧化法,设定不同氧化电压,在Ti2448合金表面生成具有不同纳米管径的生物活性膜。通过MTT实验检测不同管径纳米管表面在不同时间点黏附的MG-63细胞数量差异;通过免疫荧光法观察接种24h后不同管径纳米管表面黏附细胞形态学差异。结果显示,细胞接种3h和24h后,30nm组的OD值显著高于对照组及其他实验组(P0.05);随着纳米管径的增加,细胞骨架蛋白的铺展范围逐渐减小,细胞由多角形渐变为梭形、椭圆形、圆形。研究结果表明,30nm管径生物活性膜更有利于成骨细胞的早期黏附,可能更能抵抗细菌在钛种植体上的黏附与定植。     

Structural diversity of Papaver somniferum L. cell surfaces in vitro depending on particular steps of plant regeneration and morphogenetic program

Culture of Papaver somniferum in vitro was used for a characterisation of cell surface structures and mode of cell adhesion and cell separation during cell differentiation and plant regeneration in somatic embryogenesis and shoot organogenesis. In early stages of somatic embryogenesis, cell type-specific and developmentally regulated change of cell morphogenesis was demonstrated. Cell wall of separated embryonic cells were self-covered with external tubular network, whereas morphogenetic co-ordination of adhered cells of somatic proembryos was supported by fine and fibrillar external cell wall continuum of peripheral cells, interconnecting also local sites of cell separation. Such type of cell contacts disappeared during histogenesis, when the protodermis formation took place. Tight cell adhesion of activated cells with polar cell wall thickening, and production of extent mucilage on the periphery were the crucial aspects of meristemoids. Fine amorphous layer covered developing shoot primordia, but we have not observed such comparable external fibrillar network. On the contrary intercellular separation of differentiated cells in regenerated organs, and accepting distinct developmental system of somatic embryogenesis and shoot organogenesis, cell adhesion in early stages and ultrastructural changes associated with tissue disorganisation, and the subsequent reorganisation into either embryos or shoots appear to be regulatory morphogenetical events of plant regeneration in vitro.