The influence of structure of the interface and interphase on paint adhesion

It has been demonstrated earlier that significant adhesion enhancement to chemically inert polyolefins can be attained through surface grafted connector molecules reactive with oxidized substrate surface. The effectiveness of adhesion improvement through such tethered interfaces was shown to depend on the mode of interaction with the adjacent medium: interpenetration or chemical reaction, as well as surface density and length of grafted molecules. We have frequently observed that some systems, such as in painted products, fail through the delamination of the coating from the substrate surface at the stress levels well below the anticipated load-bearing capacity of the tethered interface. Two interim hypotheses have been formulated to explain the observed phenomenon: (i) The chain scission in surface oxidized polyolefins takes place not only in the uppermost polymer surface, but may propagate into the sub-surface region, thus creating a weak boundary layer which fails cohesively through its bulk, (ii) In order to increase the load-bearing capacity of the interphase, the sub-surface region of the substrate needs to be reinforced by short-chain molecules penetrating into and subsequently providing effective crosslinks between individual fragments of excessively oxidized and hence, weaker sub-surface part of the interphase. In this paper we verify the above hypotheses. The oxidized sub-surface layer reinforced by polyethyleneimine becomes an integral part of the effective interphase in addition to the tethered interface and the interpenetrated network of connector molecules and the paint.     

Quantitative immunofluorescent assay of full-length,recombinant CD4 in solution and mapping of its epitopes

A specific, rapid, and sensitive method for the detection of CD4 in solution was developed using pairs of fluorescently stained monoclonal antibodies which do not cross-compete. The assay is quantitated by flow cytometry using Simply Cellular microbeads (SC beads) as the primary support for the first anti-CD4 mAb. This method uses the standard conditions for anti-CD4 monoclonal antibody binding, washing, detection, and quantitation by flow cytometry of the CD4 antigen either bound to the SC beads or expressed on the cell surface. The monoclonal antibody used (Leu 3a PE) is the standard reference used to evaluate the CD4 concentration. This method differs from ELISA techniques, which need an antigen standard curve and thus can be influenced by the quality and source of the antigen. This type of assay is also a procedure which enables determination of the level of oligomerization of the bound antigen. It can be used for any antigen to which monoclonal antibodies recognizing at least two distinct epitopes are available. The use of soluble or full-length CD4 derivatives as potential therapeutic agents against AIDS, would benefit from a precise quantitation of the CD4 molecules which still have their proper tertiary structure.     

Strategies and results of atomic force microscopy in the study of cellular adhesion

Atomic Force Microscopy (AFM) provides a range of strategies for investigating living cell adhesion to the extracellular matrix, other cells or biomaterials in their native environment. This review surveys the results obtained from major studies using AFM for mechanical force evaluation in the cell, morphological visualization of the cell and studies of the cell's response to chemical or mechanical stress. Recently, the use of AFM has been broadened to obtain experimental information about cell adhesion molecules. Quantitative measurements of binding forces between adhesion proteins and their ligands in the cell or on a surface are presented. These analyses provide data on individual molecules and their resulting collective behaviour at the cell level. They significantly contribute to the characterisation of cellular adhesion with physical principles relating to biochemistry.     

Expression of classic cadherins and δ-protocadherins in the developing ferret retina

Background  

Cadherins are a superfamily of calcium-dependent adhesion molecules that play multiple roles in morphogenesis, including proliferation, migration, differentiation and cell-cell recognition. The subgroups of classic cadherins and δ-protocadherins are involved in processes of neural development, such as neurite outgrowth, pathfinding, target recognition, synaptogenesis as well as synaptic plasticity. We mapped the expression of 7 classic cadherins (CDH4, CDH6, CDH7, CDH8, CDH11, CDH14, CDH20) and 8 δ-protocadherins (PCDH1, PCDH7, PCDH8, PCDH9, PCDH10, PCDH11, PCDH17, PCDH18) at representative stages of retinal development and in the mature retina of the ferret by in situ hybridization.     

Quantum dot labeling based on near-field optical imaging of CD44 molecules

The lateral organization of membrane proteins and lipids domains has a direct impact on many cellular processes, but generally these domains are too small to be resolved by diffraction-limited resolution of fluorescence microscopy. Here, we use quantum dot (QD) labeling based on near-field optical imaging, to study the nanoscale organization of hyaluronan receptor CD44 molecules of fixed mesenchymal stem cells (MSCs) in air, with a optical resolution down to 50 nm. The photostability and high luminance of QD evidently improve the signal-to-noise ratio and reproducibility of near-field optical data. Importantly, the blinking-intensity analysis was proposed to identify single QD, providing a calibration to relate intensity to numbers of antibody for the first time. Additionally, the fluorescence-topographic imaging enables us to investigate the topographic location pattern. Our results demonstrate that CD44 molecules on MSCs are enriched into nanosized domain and they predominantly locate on the peak of the membrane protrusions, which may contribute to clarify the underlying mechanism of functions ascribed to these molecules.     

The effect of native vowel processing ability and frequency discrimination acuity on the phonetic training of English vowels for native speakers of Greek

The perception and production of nonnative phones in second language (L2) learners can be improved via auditory training, but L2 learning is often characterized by large differences in performance across individuals. This study examined whether success in learning L2 vowels, via five sessions of high-variability phonetic training, related to the learners' native (L1) vowel processing ability or their frequency discrimination acuity. A group of native speakers of Greek received training, while another completed the pre-/post-tests but without training. Pre-/post-tests assessed different aspects of their L2 and L1 vowel processing and frequency acuity. L2 and L1 vowel processing were assessed via: (a) Natural English (L2) vowel identification in quiet and in multi-talker babble, and natural Greek (L1) vowel identification in babble; (b) the categorization of synthetic English and Greek vowel continua; and (c) discrimination of the same continua. Frequency discrimination acuity was assessed for a nonspeech continuum. Frequency discrimination acuity was related to measures of both L1 and L2 vowel processing, a finding that favors an auditory processing over a speech-specific explanation for individual variability in L2 vowel learning. The most efficient frequency discriminators at pre-test were also the most accurate both in English vowel perception and production after training.     

BSRF圆二色谱研究进展

圆二色（ＣＤ）谱是一种特殊的吸收谱，它对手性分子的构象十分敏感，因此它是最重要的光谱实验之一。北京同步辐射装置（ＢＳＲＦ）３Ｂ１Ｂ生物光谱实验站已经投入运行，它提供单色聚焦的同步辐射光，波长覆盖范围１７０ｎｍ～５００ｎｍ该站的主要设备是一台圆二色谱仪，用这台仪器成功地获取了活性生物样品～捕光色素（ＬｉｇｔＨａｒｖｅｓｔＣｈｌｏｒｏｐｈｙｌｌ）的ＣＤ谱，同时测得樟脑磺酸（ＣａｍｐｈｏｒＳｕｌｆｏｎｉ     

Effects of a self-assembled monolayer on the sliding friction and adhesion of an Au surface

The friction and adhesion mechanisms with and without a self-assembled monolayer (SAM) in nanotribology were studied using molecular dynamics (MD) simulation. The MD model consisted of two gold planes with and without n-hexadecanethiol SAM chemisorbed to the substrate, respectively. The molecular trajectories, tilt angles, normal forces, and frictional forces of the SAM and gold molecules were evaluated during the frictional and relaxation processes for various parameters, including the number of CH₂ molecules, the interference magnitude, and whether or not the SAM lubricant was used. The various parameters are discussed with regard to frictional and adhesion forces, mechanisms, and molecular or atomic structural transitions. The stick–slip behavior of SAM chains can be completely attributed to the van der Waals forces of the chain/chain interaction. When the number of CH₂ molecules was increased, the SAM chains appeared to have bigger tilt angles at deformation. The magnitude of the strain energy that was saved and relaxed is proportional to the elastic deformable extent of the SAM molecules. The frictional force was higher for long chain molecules. With shorter SAM molecules, the adhesion force behavior was more stable during the compression and relaxation processes. A surface coated with a SAM can increase nano-device lifetimes by avoiding interface effects like friction and adhesion. PACS 52.65.Yy; 81.40.Pq; 81.16; 68.35.-p     

Surface Enhanced Raman Scattering on Self-assembled Nano Silver Film Prepared by Electrolysis Method

用一种廉价的电解方法制备了纳米银膜,并详细研究了在这种银膜上的表面增强拉曼散射效果．结晶紫为本实验的检测性分子．通过实验发现,这种银膜用便携式拉曼光谱仪测试并计算出的表面增强拉曼散射的增强因子为603,并对结晶紫的最小检出限为0.1 nmol/L     

Rodlike polymer gelatination as studied by PFG NMR: Fibrin polymerization in human plasma

The structure formation in rodlike polymers was investigated by the example of polymerization of fibrin rodlike, molecules in native plasma. Translational mobility of fibrin, molecules in anticoagulated plasma and native plasma during fibrin polymerization was studied by pulse field gradient nuclear magnetic resonance. It was shown that the diffusion decay of anticoagulated plasma can be fitted by the sum of exponents and fibrin molecules have the self-diffusion coefficientD _f of 1.53·10^?11 m²/s. The diffusion decay of native plasma during fibrin polymerization becomes nonexponential and is described by the lognormal distribution of fibrin self-diffusion coefficients. Qualitative and quantitative changes of the spectrum of fibrin self-diffusion coefficients (SDCs) during polymerization were investigated and analyzed. A symmetrical broadening of the spectrum at the beginning of polymerization and symmetrical narrowing at its final stages with conservation of the most probable SDC was explained on the basis of the hypothesis about the simultaneous action of fibrin polymerization and lyses.     

Regional and modular expression of morphogenetic factors in the demosponge Lubomirskia baicalensis

Some sponges [phylum Porifera], e.g. the demosponges Lubomirskia baicalensis or Axinella polypoides, show an arborescent growth form. In the freshwater sponge L. baicalensis this morphotype is seen mostly in depths below 4m while in more shallow regions it grows as a crust. The different growth forms are determined in nature very likely by water current and/or light. The branches of this species are composed of modules, arranged along the apical-basal axis. The modules are delimited by a precise architecture of the spicule bundles; longitudinal bundles originate from the apex of the earlier module, while at the basis of each module these bundles are cross-linked by traverse bundles under formation of annuli. Genes encoding putative morphogenetic factors, myotrophin and epidermal growth factor (EGF)-like molecules, and one gene of an antagonist for the Wnt signaling pathway, the soluble frizzled molecule, have been identified and characterized. Their expression levels as well as those of silicatein, one major spicule-forming molecule, have been studied in the crusts and the modules. The data revealed that at the apices of each module higher level of expression of myotrophin and EGF can be detected, while the base of each module is characterized by a high steady-state expression level of soluble frizzled molecule. These results suggest that module formation in L. baicalensis is controlled by a tuned interaction of agonistic (e.g., myotrophin and EGF) as well as antagonistic morphogenetic factors (e.g., soluble frizzled molecule).     

Quantifying the intelligibility of speech in noise for non-native talkers

The intelligibility of speech pronounced by non-native talkers is generally lower than speech pronounced by native talkers, especially under adverse conditions, such as high levels of background noise. The effect of foreign accent on speech intelligibility was investigated quantitatively through a series of experiments involving voices of 15 talkers, differing in language background, age of second-language (L2) acquisition and experience with the target language (Dutch). Overall speech intelligibility of L2 talkers in noise is predicted with a reasonable accuracy from accent ratings by native listeners, as well as from the self-ratings for proficiency of L2 talkers. For non-native speech, unlike native speech, the intelligibility of short messages (sentences) cannot be fully predicted by phoneme-based intelligibility tests. Although incorrect recognition of specific phonemes certainly occurs as a result of foreign accent, the effect of reduced phoneme recognition on the intelligibility of sentences may range from severe to virtually absent, depending on (for instance) the speech-to-noise ratio. Objective acoustic-phonetic analyses of accented speech were also carried out, but satisfactory overall predictions of speech intelligibility could not be obtained with relatively simple acoustic-phonetic measures.     

The biological characteristics of different generation mesenchymal stem cells cultured in vitro and cocultured with vascular scaffold

Mesenchymal stem cells (MSCs) were used widely as seed cells in tissue engineering blood vessel construction. However, the biological characteristics difference of different generation MSCs in vitro culture is unknown, which laid a foundation for appropriate generation seeded cells selection for tissue engineering blood vessel construction. In this report, MSCs were isolated from SD rat bone marrow and identified by flow cytometry; cell growth curve test, cell surface antigen expression rate detection, cryopreservation resuscitation rate test, CD31 expression rate test, cell cycle analysis, and adhesion difference on vascular scaffold test were performed. The research results indicated that the MSCs shape was spindle and uniform with vigorous growth. CD105 and CD90 factor expression rate reached 82.5 and 84.9%, respectively, and the expression rate of CD45 was only 7.3%. The proliferation capacity of the fourth generation MSCs were more exuberant, with proliferation index as 20.3%; the cell proliferation index of the eighth generation decreased to only 9.1%. The cryopreservation resuscitation rate of the second generation and fourth generation MSCs were both higher than 80%, and the cryopreservation resuscitation rate of the eighth generation MSCs was only about 60%. After the induction for 5 days, MSCs had weak CD31 expression, and with the prolonged induction time, expression increased. All generation MSCs expressed CD31 after being induced for 10 days; however, the CD31 positive expression rate of the second generation, fourth generation, and sixth generation MSCs had significant difference with the eighth generation MSCs. Adhesion rate of MSCs before sixth generation was around 40%, but the adhesion rate of eighth generation MSCs was only about 27%. In all, biological characteristics of different generation MSCs existed certain differences, and especially the eighth generation MSCs aged seriously, whose cell activity decreased significantly. The researchers believed that the MSCs before the sixth generation can maintain excellent properties of MSCs, and can be used as seed cells for vascular tissue engineering.     

Using self-assembled monolayer technology to probe the mechanical response of the fiber interphase-matrix interphase interface

In this paper, a brief review of the fiber-matrix interphase/interface region is given for carbon- and glass-fiber composites. The substructure of the interphase/interface region is discussed in terms of three interphases: (a) the fiber interphase (FI), (b) the sizing interphase (SI), and (c) the matrix interphase (MI), and two interface regions: (a) the FI-SI interface and (b) the SI-MI interface. These substructures are a synthesis of the ideas advanced by Ishida and Koenig and Drzal. The schematic model of interphase deformation behavior originally given by Bascom is reconstructed to include research results from the above researchers. To systematically probe adhesion at the SI-MI interface, functionalized self-assembled monolayers (SAMs) using bonding and non-bonding C₁₁- type trichlorosilanes are prepared using the research of Menzel and Heise, and that of Cave and Kinloch as a guide. Results from this research are compared with short chain bonding and nonbonding silanes prepared by aqueous and non-aqueous deposition processes. The data were interpreted using the mechanisms proposed by Sharpe, Ishida and Koenig, and Drzal and the mathematical equation proposed by Nardin and Ward. For the non-bonding short-chain silane deposited by aqueous deposition, 90% of the adhesion was found to be due to mechanical interlocking, with the remaining adhesion due to physicochemical interactions. For the bonding short-chain silane deposited by aqueous deposition, the interface strength relative to the non-bonding short-chain silane increased by 31%. However the interfacial shear strength (IFSS) of this system was approximately 40% lower than the comparable bonding SAM interface. This difference was interpreted in terms of the propensity of the C₃-alkylamine to form cyclic ring structures in the MI region as described by Ishida, Koenig, et al. The SAM data also indicates that 70-85% of the maximum IFSS is obtained with 25-50% of the surface covered with functional groups. This suggests that steric hindrance, due to the size of the DGEBA molecules, restricts access to the functional groups on the surface. Therefore, only 35% of the surface functional groups are accessible for bonding in the DGEBA/m-PDA epoxy resin system.     

High-Sensitivity Immunofluorescence Staining: A Comparison of the Liposome Procedure and the FASER Technique on mGR Detection

Flow cytometry has become a widely-used and powerful tool for the characterization of cells according to their expression of specific proteins. However, sensitivity of this method is still limited since conventionally labeled antibodies can be conjugated with at maximum 1–10 dye molecules. This fact resulted in the need to develop new techniques in order to identify molecules which are expressed in very low but functionally relevant amounts. In the past, we have successfully used a liposome-based high-sensitivity immunofluorescence technique to measure the expression of low abundant membrane bound glucocorticoid receptors (mGR) on different cell types. The use of this technique allows the detection of as few as 50–100 antigen molecules per cell which is due to a 100-fold to 1000-fold increase in fluorescence signal intensity compared with conventional methods. The higher sensitivity is achieved since thousands of dye molecules can be enclosed in liposomes. Another modern high-sensitivity immunofluorescence staining method is the purchasable Fluorescence Amplification by Sequential Employment of Reagents (FASER) procedure. Here, we aimed at comparing sensitivity and specificity of these two techniques for the detection of the mGR. Our data demonstrate the FASER technique to be more sensitive and also more specific for the detection of mGR as compared to the liposome technique. However, both methods have advantages and disadvantages which are discussed in detail.     

Research on the icephobic properties of fluoropolymer-based materials

Fluoropolymer, because of the extremely low surface energy, could be non-stick to water and thus could be a good candidate as anti-icing materials. In this paper, the icephobic properties of a series of fluoropolymer materials including pristine PTFE plates (P-PTFE), sandblasted PTFE plates (SB-PTFE), two PTFE coatings (SNF-1 and SNF-CO1), a fluorinated room-temperature vulcanized silicone rubber coating (F-RTV) and a fluorinated polyurethane coating (F-PU) have been investigated by using SEM, XPS, ice adhesion strength (tensile and shear) tests, and static and dynamic water contact angle analysis. Results show that the fluoropolymer material with a smooth surface can significantly reduce ice adhesion strength but do not show obvious effect in reducing ice accretion at −8 °C. Fluoropolymers with sub-micron surface structures can improve the hydrophobicity at normal temperature. It leads to an efficient reduction in the ice accretion on the surface at −8 °C, due to the superhydrophobicity of the materials. But the hydrophobicity of this surface descends at a low temperature with high humidity. Consequently, once ice layer formed on the surface, the ice adhesion strength enhanced rapidly due to the existence of the sub-micron structures. Ice adhesion strength of fluoropolymers is highly correlated to CA reduction observed when the temperature was changed from 20 °C to −8 °C. This property is associated with the submicron structure on the surface, which allows water condensed in the interspace between the sub-micron protrudes at a low temperature, and leads to a reduced contact angle, as well as a significantly increased ice adhesion strength.     

The platelet integrin alpha(IIb) beta(3) imaged by atomic force microscopy on model surfaces

The platelet membrane receptor alpha(IIb) beta(3) binds to adsorbed protein ligands including fibrinogen, von Willebrand factor and fibronectin, and is critically important in mediating platelet adhesion to damaged subendothelium and to synthetic biomaterial surfaces. This receptor is a member of the integrin family, a highly prevalent class of heterodimeric molecules consisting of a single alpha and beta subunit. In an ongoing effort to understand the mechanisms underlying platelet adhesion events, high-resolution atomic force microscopy (AFM) under dynamic conditions was used to obtain images of alpha(IIb) beta(3) molecules as well as aggregates of the protein. Images of integrin molecules were obtained by tapping mode AFM under aqueous buffer conditions following adsorption on a series of ultrasmooth model surfaces. On a model hydrophobic surface, detergents stabilizing the protein in solution competed for surface adsorption sites. When this detergent was removed from the system, the protein was predominantly seen as aggregates with head groups pointing outward. A limited number of individual integrin molecules were observed, and were found to have dimensions consistent with those reported previously by electron microscopy studies. Integrin molecules showed weak adhesion to the two hydrophilic surfaces used in the study, although formation of a lipid bilayer around surface-adsorbed molecules improved the resolution. At longer time periods, the integrin molecules embedded in this lipid bilayer exhibited sufficient mobility to form molecular aggregates. The structural measurements described in this study not only reveal three-dimensional features of the molecule, they represent an important step towards dynamic adsorption experiments and visualizing the integrin interacting with surface-adsorbed proteins as in biomaterial-induced thrombogenesis.     

Physics of cell elasticity,shape and adhesion

We review recent theoretical work that analyzes experimental measurements of the shape, fluctuations and adhesion properties of biological cells. Particular emphasis is placed on the role of the cytoskeleton and cell elasticity and we contrast the shape and adhesion of elastic cells with fluid-filled vesicles. In red blood cells (RBC), the cytoskeleton consists of a two-dimensional network of spectrin proteins. Our analysis of the wavevector and frequency dependence of the fluctuation spectrum of RBC indicates that the spectrin network acts as a confining potential that reduces the fluctuations of the lipid bilayer membrane. However, since the cytoskeleton is only sparsely connected to the bilayer, one cannot regard the composite cytoskeleton–membrane as a polymerized object with a shear modulus. The sensitivity of RBC fluctuations and shapes to ATP concentration may reflect topological defects induced in the cytoskeleton network by ATP. The shapes of cells that adhere to a substrate are strongly determined by the cytoskeletal elasticity that can be varied experimentally by drugs that depolymerize the cytoskeleton. This leads to a tension-driven retraction of the cell body and a pearling instability of the resulting ray-like protrusions. Recent experiments have shown that adhering cells exert polarized forces on substrates. The interactions of such “force dipoles” in either bulk gels or on surfaces can be used to predict the nature of self-assembly of cell aggregates and may be important in the formation of artificial tissues. Finally, we note that cell adhesion strongly depends on the forces exerted on the adhesion sites by the tension of the cytoskeleton. The size and shape of the adhesion regions are strongly modified as the tension is varied and we present an elastic model that relates this tension to deformations that induce the recruitment of new molecules to the adhesion region. In all these examples, cell shape and adhesion differ from vesicle shape and adhesion due to the presence of the elastic cytoskeleton and to the fact that active processes (ATP, molecular motors) within the cell modify cytoskeletal elasticity and tension.     

Discovery of NKT cells and development of NKT cell-targeted anti-tumor immunotherapy

Natural Killer T (NKT) cells are unique lymphocytes characterized by their expression of a single invariant antigen receptor encoded by Vα14Jα18 in mice and Vα24Jα18 in humans, which recognizes glycolipid antigens in association with the monomorphic CD1d molecule. NKT cells mediate adjuvant activity to activate both CD8T cells to kill MHC-positive tumor cells and NK cells to eliminate MHC-negative tumor at the same time in patients, resulting in the complete eradication of tumors without relapse. Therefore, the NKT cell-targeted therapy can be applied to any type of tumor and also to anyone individual, regardless of HLA type.Phase IIa clinical trials on advanced lung cancers and head and neck tumors have been completed and showed significantly prolonged median survival times with only the primary treatment. Another potential treatment option for the future is to use induced pluripotent stem cell (iPS)-derived NKT cells, which induced adjuvant effects on anti-tumor responses, inhibiting in vivo tumor growth in a mouse model.     

Spectroscopic and Electrochemical Studies on the Interaction of Carmoisine Food Additive with Native Calf Thymus DNA

ABSTRACT

In this work, for the first time interaction between a carmoisine food additive and native calf thymus DNA was monitored using UV-Vis absorption, fluorescence, and circular dichroism (CD) spectroscopy, as well as cyclic voltammetry and viscosity measurements. It can be concluded that carmoisine could interact with DNA via a groove-binding mode as evidenced by a hyperchromic effect of absorption spectra, increases in the fluorescence quenching effect of DNA, certain induced CD spectral changes, and relatively small changes in the viscosity of DNA. The binding constants (K_b) for the carmoisine with DNA was estimated to be 6.2 × 10⁴ M^?1 through spectroscopic titrations. The cyclic voltammetry method showed that both anodic and cathodic peak currents of carmoisine decreased upon addition of the DNA. Circular dichroism spectra indicated that there are certain detectable conformational changes such as conversion from B-like to A-like in the DNA double helix when carmoisine was added.