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To probe the migration of free radicals (FRs), the reduction behaviours of hexavalent chromium (Cr(VI)) in water and ice by high-energy electron beam (HEEB) irradiation were investigated. Interestingly, the reductive efficiency (RE) of Cr(VI) in water was appreciably higher than that in ice. Thus, it was proposed that the migration ability of FRs in water is distinctly higher than that in ice, likely because the migration performance of FRs is closely related to the intermolecular distance of water molecules. Furthermore, the RE of Cr(VI) in ice decreased gradually with the distance from the irradiated area, indicating that FRs could migrate in ice and that the migration performance was closely related to the RE. Additionally, FRs (hydrated electrons ([Formula: see text]) and hydrogen radicals (·H)) generated during the irradiation process played a key role in the reduction of Cr(VI). Hydroxyl radicals (·OH) and H 2 O 2 were the dominant negative factors for the reduction because of their oxidizing effects, but these factors could be eliminated by the removal of ·OH. This work reveals the migration performance of FRs in different media for the first time. This result may be useful for basic and applied studies in fields of environmental science related to FRs.

Magnetic relaxation switch demonstrated that the aggregated nanomagnetic iron oxide (NMIO) nanocrystal possessed a lower T 2 value and better relaxivity compared with monodispersed NMIO nanocrystal. However, we found that NMIO nanoclusters (NMIONCs) showed a different magnetic resonance (MR) imaging property in comparison with NMIO nanocrystals. Herein, three types of NMIONCs were used to explore the effects of size and compositions on the variations of magnetism and MR contrast ability. It was found that the transverse relaxation rate (r 2 ) of NMIONCs depended on the contact area between particles and water molecules. The smaller size and higher solubility could carry out higher contact area between NMIONCs and water molecules. Therefore, the monodispersed NMIONC showed a better T 2 contrast ability in comparison with the aggregated NMIONC. In addition, for NMIONCs with the same composition, the magnetism and contrast ability gradually increased with the particle size decreasing. In vivo, NMIONCs that possessed the best solubility and the smallest size showed the most effective MR contrast effect for the liver region of mice. As a result, the size and composition of NMIONCs played important roles for enhancing contrast behavior. This study provides a new idea to develop high-performance T 2 contrast agents by modulating the size and composition of particles.

Considering the chemical exchange between gadolinium centers and water protons, nanosystems comprising gadolinium conjugated with high specific area nanocarriers might serve as more robust clinical tools for diagnosis and imaging-guided therapy. Herein, a pH-responsive nanosystem containing graphene oxide conjugated with a folic acid- and gadolinium-labeled dendrimer (FA-GCGLD) to boost its T 1 contrast ability was developed, and doxorubicin (DOX) and colchicine (COLC) were efficiently loaded onto this nanosystem (FA-GCGLD-DOX/COLC). This nanosystem showed a prominent T 1 contrast with an ultrahigh relaxivity of up to 11.6 mM -1 s -1 and pH-responsive drug release behavior. HepG2 cells treated with FA-GCGLD-DOX/COLC were efficiently inhibited, and the cell contrast was enhanced. In vivo, the tumor accumulation of FA-GCGLD-DOX/COLC significantly increased, thereby facilitating the systemic delivery of particles and exerting tumor growth inhibition and an enhanced tumor contrast effect. Moreover, compared to free drugs, FA-GCGLD-DOX/COLC effectively decreased the drug resistance of the tumor, thereby improving the cancer chemotherapeutic efficacy. In addition, injecting rats with FA-GCGLD afforded excellent magnetic resonance angiography (MRA) images with high-resolution vascular structures because of the long blood circulation time of FA-GCGLD. Thus, this study provides a powerful tool for diverse applications in the biomedical field, including accurate diagnosis and chemotherapy of tumors and the detection of cardiovascular diseases.

A wormstructured and nanosized porous/hollow polyethyleneimine (PEI) functionalized Gd2O3/Fe3O4 composite was fabricated as a drug carrier and a bioimaging agent. The effect of PEI's chain length on the size and morphology of the nanoworm was investigated and the results indicated that the nanoworm modified with PEI (10,000 molecular weight) (designated as p-nanoworm) possessed a suitable size and a porous/hollow structure. Meanwhile, the p-nanoworm could effectively prevent the leakage of Gd ions under different pH conditions because of plenty of amino groups on their surface. Compared with contrast agents of clinical use, the p-nanoworm displayed MR enhancement with a high r1 relaxivity of 5.58 s(-1) mM(-1) per gadolinium atom. Cisplatin (CDDP), a clinical anticancer drug, could be easily loaded into the pores and lumen of the p-nanoworm (p-nanoworm-CDDP) and also controllably released by adjusting the pH value. Cell assay suggested that the p-nanoworm possessed satisfactory biocompatibility and meanwhile could promote CDDP uptake of HeLa cells and enhance the inhibition effect on HeLa cells. In addition, p-nanoworm-CDDP showed a negligible cytotoxicity on normal human cells, indicating that the side effect of CDDP is reduced. Thus, the p-nanoworm could have a potential application for the diagnosis and therapy of cancer.

Pistacia chinensis Bunge (P. chinensis) is a deciduous and dioecious perennial arbor of the family Anacardiaceae that flowers from March to April and bears fruit from September to October. There are three rapidly growing stages in the annual growth process of P. chinensis. However, the knowledge of the secondary metabolites related to P. chinensis gender and growth season remains scant. In this study, HPLC was used to qualitatively and quantitatively determine the content of the catechin hydrate, rutin, quercetin, and kaempferol contents in male and female tree inflorescences and leaves. Total phenolics and flavonoids were also detected using a spectrophotometer. The results indicated that the contents of these compounds fluctuated with seasons and they reached the highest levels in nascent leaves. The fluctuations of these compounds followed different pathways of evolution, by increasing or decreasing in male and female trees throughout the whole growth process because they had their own biological functions. Moreover, the extracts exhibited DPPH radical scavenging bioactivity and showed no significant cytotoxicity towards 3T3-L1 preadipocytes. Together, these results demonstrated that P. chinensis has great potential as an antioxidant medicine, and the best harvest time is in the spring., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

In this work, a long-duration remediation agent (LRA) on hexavalent chromium (Cr(VI)) was developed using sodium thiosulfate (ST) supported by attapulgite (ATP) micro-nano networks induced through high-energy electron beam (HEEB) irradiation. The ATP networks could effectively reduce the leaching amount of Cr(VI) in soil. More importantly, the ATP networks could significantly control the leaching behavior of ST, and then prolong the duration and increase the reduction efficiency of ST on Cr(VI). As a result, LRA displayed high performance on controlling the migration of Cr(VI) and reducing Cr(VI) to Cr(III). Additionally, pot experiment indicated that LRA could effectively decrease the absorbed amount of Cr(VI) in corn, and reduce the inhibition effect of Cr(VI) on the growth of corn. Therefore, this work could provide a facile approach to remediate the Cr(VI)-contaminated soil and lower the harmful effect of Cr(VI) on crop., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We developed a highly efficient magnetic relaxation switch (MRS) system based on poly(vinyl alcohol) functionalized nanomagnetic iron oxide (PVA@NMIO) particles for the detection of boric acid or borate ester (BA/BE). It was found that the addition of BA/BE induced the aggregation of PVA@NMIO particles, resulting in a measurable change in the T2 relaxation time in magnetic resonance measurements. The main mechanism was proposed that the electron-deficient boron atoms of BA/BE caused the aggregation of PVA@NMIO particles through covalent binding to the hydroxyl groups of PVA. This novel detection system displayed excellent selectivity, high sensitivity, and rapid detection for BA/BE. Thus, this system may provide a great application prospect for detection of BA/BE.

Urea was used to disperse nanoscale zero-valent iron (NZVI) for reduction of Cr(VI) to Cr(III) in aqueous solution. Scanning electron microscope and fourier transform-infrared spectra investigations demonstrated that urea could effectively increase the dispersion of NZVI resulting in more effective reduction sites (ERS) for Cr(VI) ions. Batch reduction experiments indicated that the reductive capacity of urea dispersed NZVI (UNZVI) was significantly improved, as the reductive efficiency reached 96.8% under optimal condition compared with the raw NZVI (72.14%). Additionally, the NZVI was stable for at least 28 days after urea treatment. The dispersion mechanism was proposed that the steric hindrance effect of the urea coating on the surface might play a key role in dispersing the NZVI particles.

A pH-responsive nanoplatform, hydroxylated mesoporous nanosilica (HMNS) coated by polyethylenimine (PEI) coupled with gadolinium and folic acid (FA) (Gd-FA-Si), was designed to deliver anticancer drug targeting and to promote contrast effect for tumor cells using magnetic resonance (MR) spectrometer. Doxorubicin (DOX) was chosen as the anticancer drug and loaded into nanopores of HMNS, then its release in simulated body fluid could be controlled through adjusting the pH. This nanoplatform could significantly enhance the MR contrast effect, and the highest theoretical relaxivity per nanoplatform could even be approximately 1.28 × 10(6) mm(-1)s(-1) because of the high Gd payload (2.61 × 10(5) per nanoplatform). The entire system possessed a high targeting performance to Hela and MDA-MB-231 cells because the FA located in the system could specifically bind to the folate-receptor sites on the surface of cell. Compared with free DOX, the nanoplatform presented a higher cell inhibition effect on the basis of cell assay. Therefore, this nanoplatform could be potentially applied as a tumor-targeted T1 MR contrast agent and pH-sensitive drug carrier system.

This work developed an immediate and high-performance remediation method for Cr(VI) contaminated soil (CCS) using high energy electron beam (HEEB) irradiation. The result indicated that, compared with γ-ray irradiation, HEEB irradiation displayed a significant reduction efficiency on Cr(VI) in CCS to Cr(III) with substantially lower toxicity, which was mainly attributed to the reduction effects of electrons, hydrated electrons, and reductive radicals generated in the irradiation process of HEEB. This work could provide a one-step and effective method for the remediation of heavy metal contaminated soil (HMCS)., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Low pH triggers the translocation domain of diphtheria toxin (T-domain), which contains 10 α helices, to insert into a planar lipid bilayer membrane, form a transmembrane channel, and translocate the attached catalytic domain across the membrane. Three T-domain helices, corresponding to TH5, TH8, and TH9 in the aqueous crystal structure, form transmembrane segments in the open-channel state; the amino-terminal region, TH1-TH4, translocates across the membrane to the trans side. Residues near either end of the TH6-TH7 segment are not translocated, remaining on the cis side of the membrane; because the intervening 25-residue sequence is too short to form a transmembrane α-helical hairpin, it was concluded that the TH6-TH7 segment resides at the cis interface. Now we have examined this segment further, using the substituted-cysteine accessibility method. We constructed a series of 18 mutant T-domains with single cysteine residues at positions in TH6-TH7, monitored their channel formation in planar lipid bilayers, and probed for an effect of thiol-specific reagents on the channel conductance. For 10 of the mutants, the reagent caused a change in the single-channel conductance, indicating that the introduced cysteine residue was exposed within the channel lumen. For several of these mutants, we verified that the reactions occurred primarily in the open state, rather than in the flicker-closed state. We also established that blocking of the channel by an amino-terminal hexahistidine tag could protect mutants from reaction. Finally, we compared the reaction rates of reagent added to the cis and trans sides to quantify the residue's accessibility from either side. This analysis revealed abrupt changes in cis- versus trans-side accessibility, suggesting that the TH6-TH7 segment forms a constriction that occupies a small portion of the total channel length. We also determined that this constriction is located near the middle of the TH8 helix., (© 2015 Kienker et al.)

Traditional pesticides (TP) often do not adhere tightly to crop foliage. They can easily enter the surrounding environment through precipitation and volatilization. This can result in the pollution of the surrounding soil, water, and air. To reduce pesticide pollution, we developed a loss-control pesticide (LCP) by adding attapulgite with a nano networks structure fabricated using high energy electron beam (HEEB) irradiation and hydrothermal treatment to TP. HEEB irradiation effectively dispersed originally aggregated attapulgite through modified thermal, charge, and physical effects. Hydrothermal treatment further enhanced the dispersion of attapulgite to form nano porous networks via thermal and wet expansion effects, which are beneficial for pesticide binding. An LCP has improved retention on crop leaf surfaces. It has a higher adhesion capacity, reduced leaching and volatilization, and extended residual activity compared with the TP formulation. The treatment increases the residual activity of pesticides on crop foliage and decreases environmental pollution.

Raman spectroscopy and nonisothermal thermogravimetric analysis (TGA) measurements have been reported for different rank coals (lignite, bituminous coal, and anthracite) and the relationship between the measurements was examined. It was found that the Raman spectra parameters can be used to characterize structure changes in the different rank coals, such as the band area ratios based on the curve-fitted results. Higher ranked coal was found to have higher values of I GR/I All and I (G + GR)/I All but lower values of I D/I (G+GR), I DL/I (G+GR), I (S + SL)/I (G+GR), and I (GL+GL')/I (G+GR). The oxidation properties of the coal samples were characterized by the reactivity indexes T ig, T 20%, and T max from TGA data which were found to correlate well with the band area ratios of I GR/I All, I (G + GR)/I All, and I (S + SL)/I (G+GR). Based on these correlations, the Raman band area ratios were found to correlate with the oxidation activity of coal providing additional structural information which can be used to understand the changes in the TGA measurements.

An efficient enzyme-responsive controlled release carrier system was successfully fabricated using single-stranded DNA encapsulated functional mesoporous silica nanoparticles. Mesoporous silica nanoparticles were initially fabricated through hydrolysis of tetraethyl orthosilicate (TEOS) in cetyltrimethylammonium chloride (CTAC) solution, and the surface of nanoparticles could be encapsulated with single-stranded DNA. This nanomaterial has excellent bioactivity and its hydrolysate cannot cause damage to normal cell, thus the biocompatibility of the nanomaterial is improved. In addition, this nanomaterial showed an excellent drug release performance when loaded with drugs, which would be helpful to increase the treatment efficiency and decrease side effects of drugs.

An electrochemical method based on square wave anodic stripping voltammetry (SWASV) was developed to detect the apoptosis of yeast cells conveniently and quantitatively through the high affinity between Cu(2+) and phosphatidylserine (PS) translocated from the inner to the outer plasma membrane of the apoptotic cells. The combination of negatively charged PS and Cu(2+) could decrease the electrochemical response of Cu(2+) on the electrode. The results showed that the apoptotic rates of cells could be detected quantitatively through the variations of peak currents of Cu(2+) by SWASV, and agreed well with those obtained through traditional flow cytometry detection. This work thus may provide a novel, simple, immediate and accurate detection method for cell apoptosis.

Nitrogen fertilizer unabsorbed by crops eventually discharges into the environment through runoff, leaching and volatilization, resulting in three-dimensional (3D) pollution spanning from underground into space. Here we describe an approach for controlling nitrogen loss, developed using loss control fertilizer (LCF) prepared by adding modified natural nanoclay (attapulgite) to traditional fertilizer. In the aqueous phase, LCF self-assembles to form 3D micro/nano networks via hydrogen bonds and other weak interactions, obtaining a higher nitrogen spatial scale so that it is retained by a soil filtering layer. Thus nitrogen loss is reduced and sufficient nutrition for crops is supplied, while the pollution risk of the fertilizer is substantially lowered. As such, self-fabrication of nano-material was used to manipulate the nitrogen spatial scale, which provides a novel and promising approach for the research and control of the migration of other micro-scaled pollutants in environmental medium.

Vermiculite and vermiculite modified with hydrochloric acid were investigated to evaluate their flocculation efficiencies in freshwater containing harmful algae blooms (HABs) (Microcystis aeruginosa). Scanning electron microscope, Fourier transform infrared spectroscopy, X-ray diffraction, converted fluorescence microscope, plasma-atomic emission spectrometry, and Zetasizer were used to study the flocculation mechanism of modified vermiculite. It was found that the vermiculite modified with hydrochloric acid could coagulate algae cells through charge neutralization, chemical bridging, and netting effect. The experimental results show that the efficiency of flocculation can be notably improved by modified vermiculite. Ninety-eight per cent of algae cells in algae solution could be removed within 10 min after the addition ofmodified vermiculite clay. The method that removal of HABs with modified vermiculite is economical with high efficiency, and more research is needed to assess their ecological impacts before using in practical application.

High energy electron beam (HEEB) irradiation was used to disperse nanoscale zero-valent iron (NZVI) for reduction of CrVI to CrIII in aqueous solution. Pore size distribution, scanning electron microscopy and X-ray diffraction characterizations demonstrated that HEEB irradiation could effectively increase the dispersion of NZVI resulting in more active reduction sites of Crvi on NZVI. Batch reduction experiments indicated that the reductive capacity of HEEB irradiation-modified NZVI (IMNZVI) was significantly improved, as the reductive efficiency reached 99.79% under the optimal conditions (electron beam dose of 30 kGy at 10 MeV, pH 2.0 and 313 K) compared with that of raw NZVI (72.14%). Additionally, the NZVI was stable for at least two months after irradiation. The modification mechanism of NZVI by HEEB irradiation was investigated and the results indicated that charge and thermal effects might play key roles in dispersing the NZVI particles.

Pesticide sprayed onto plant leaves tends to discharge into the environment through rainwater washing, leaching, and volatilization, resulting in severe pollution to soil, water, and air. Here, to control pesticide loss, we developed a loss-control pesticide (LCP) by adding straw ash-based biochar and biosilica (BCS) to traditional pesticide. BCS possesses a porous micro/nano structure and thus can adsorb a large amount of pesticide molecules to form pesticide-BCS complexes that tend to be retained by the rough surface of plant leaves, displaying a high adhesion performance on the leaves; therefore, the pesticide loss decreases, sufficient pesticide for the plant is supplied, and the pollution risk of the pesticide can be substantially lowered.

A new kind of functional graphene oxide with fine stability in water was fabricated by mixing graphene oxide (GO) and brilliant blue (BB) with a certain weight ratio. The adsorption performance of this mixture of BB and GO (BBGO) to polycyclic aromatic hydrocarbons (anthracenemethanol (AC) and fluoranthene (FL)) was investigated, and the results indicated BBGO possessed adsorption capacity of 1.676 mmol/g and removal efficiency of 72.7% as to AC and adsorption capacity of 2.212 mmol/g and removal efficiency of 93.2% as to FL. After adsorption, pH and temperature-sensitive coagulation (PTC) method was used to remove the AC/BBGO or FL/BBGO complex and proved to be an effective approach to flocculate the AC/BBGO or FL/BBGO complex into large flocs, which tended to be removed from the aqueous solution.

Pesticide sprayed onto crop leaves tends to be washed off by rainwater and discharge into the environment through leaching and runoff, resulting in severe pollution to both soil and water. Here, to control pesticide loss, we developed a loss-control pesticide (LCP) by adding modified natural nanoclay (diatomite) through high-energy electron beam (HEEB) to traditional pesticide. After HEEB treatment, the originally clogged pores in diatomite opened, resulting in plenty of micro-nanopores in diatomite, which are beneficial for the pesticide molecules to access and be adsorbed. This pesticide-diatomite complex tended to be retained by the rough surface of crop leaves, displaying a high adhesion performance onto the leaves, so that the pesticide loss reduced, sufficient pesticide for crops was supplied, and the pollution risk of the pesticide could be substantially lowered.

A novel strategy to fabricate hydrogen peroxide (H(2)O(2)) sensor was developed by electrodepositing Ag nanoparticles (NPs) on a glassy carbon electrode modified with natural nano-structure attapulgite (ATP). The result of electrochemical experiments showed that such constructed sensor had a favorable catalytic ability to reduce H(2)O(2). The good catalytic activity of the sensor was ascribed to the ATP that facilitated the formation and homogenous distribution of small Ag NPs. The resulted sensor achieved 95% of the steady-state current within 2s and had a 2.4 μM detection limit of H(2)O(2)., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Natural attapulgite (N-AT) and modified attapulgite (M-AT) were used in this study to evaluate their flocculation efficiencies and mechanisms in freshwater containing harmful algal blooms through conventional jar test procedure. The experimental results showed that the efficiency of flocculation can be significantly improved by M-AT under appropriate conditions. It was found that the attapulgite modified by hydrochloric acid was similar to polyaluminum ferric silicate chloride (PAFSiC). The high efficiency for M-AT to flocculate Microcystis aeruginosa in freshwater was due to the mechanism of bridging and netting effect. Caenorhabditis elegans was used to detect the toxicity of N-AT and M-AT. The results showed that there was no significant toxicity on this organism. Attapulgite is a natural material, which can be readily available, abundant, and relatively inexpensive. Using modified attapulgite to remove the harmful algal blooms could have the advantages of high effectiveness, low cost, and low impact on the environment., (Copyright © 2011. Published by Elsevier Ltd.)

A novel third-generation hydrogen peroxide (H(2)O(2)) biosensor was developed by immobilizing horseradish peroxidase (HRP) on a biocompatible attapulgite (ATP) modified glassy carbon (GC) electrode. The ATP could provide a biocompatible microenvironment for enzyme molecules, greatly amplify the coverage of HRP molecules on the electrode surface, and most importantly facilitate the direct electron transfer between HRP and the electrode. The biosensor construction process was followed by atomic force microscopy (AFM). Cyclic voltammetry was employed to characterize the properties of the biosensor. A linear calibration plot of the enzyme electrode was obtained over the range of 5 µM to 0.3 mM for H(2)O(2) with a detection limit of 5 µM. Furthermore, the biosensor showed high sensitivity, good reproducibility, and fine long-term stability., (2011 © The Japan Society for Analytical Chemistry)

An inorganic nano-network of attapulgite is formed from rigid nanorods using ion beam bombardment. The structure of the nano-networks depends on the ion beam fluence for the same ion energy. Scanning electron microscopy reveals that ion beam bombardment improves the dispersion of the attapulgite particles and the change in the shape of the rod-shaped attapulgite particles stems from the thermal stress induced by ion beam bombardment. This phenomenon is more obvious for higher ion fluences. The bent or twisted rod-shaped attapulgite particles cross-link to form a network structure, which is stable in water, and when the ion fluence is increased further, the cross-linked points are permanently sealed. The improved materials are more useful than clava attapulgite particles.

Objective: Radiation can activate the phosphatidylinositol 3-kinase/Akt pathway and cannot downregulate survivin expression in breast cancer cells. Deguelin induces apoptosis in breast cancer cells by inhibiting pAkt and survivin expression. In this study, we assessed the effect of deguelin on radiosensitization of human breast cancer cells and its possible mechanism., Methods: Deguelin and radiation were administrated in MDA-MB-231 human breast cancer cells. The cytotoxic interactions and mutual influences between these 2 modalities were analyzed by a series of assays including clonogenic, flow cytometric, and Western blotting., Results: Phospho-Akt expression and survivin expression significantly decreased after 10 nM deguelin treatment, while phospho-Akt expression increased and survivin expression did not alter after radiation (3 Gy). However, phospho-Akt expression and survivin expression further significantly decreased after deguelin combined with radiation treatment. Deguelin combined with radiation markedly decreased clonogenic cell survival. After treatment of deguelin (10 nM) combined with radiation (3 Gy), caspase-dependent apoptosis was significantly increased and cell cycle was arrested in the G2-M phase in MDA-MB-231 cells., Conclusions: Deguelin attenuates radiation-induced prosurvival Akt signaling and enhances the radiosensitivity of MDA-MB-231 cells, and the mechanisms for this action may include inhibiting phospho-Akt and survivin expression, increasing caspase-dependent apoptosis, and prolonging cell cycle arrest in the G2-M phase.

Polyprenyl phosphates, as well as polyprenyl alcohols bearing different isopentenyl C(5) units, have been synthesized. The pH range of spontaneous vesicle formation of polyprenyl phosphates with or without polyprenyl alcohols was defined by fluorescence microscopy. A variety of the acyclic or monocyclic polyprenyl phosphates studied formed stable vesicles in water over a wide range of pHs, and the addition of polyprenyl alcohols allowed the vesicle formation of polyprenyl phosphates at higher pHs. Osmotic swelling of a suspension of unilamellar vesicles using the stopped-flow/light-scattering method enabled us to evaluate the water permeability of polyprenyl phosphate vesicles with or without 10 mol% of free polyprenyl alcohol. The addition of many polyprenyl alcohols to polyprenyl phosphate vesicles decreased the water permeability, and some reduced it even more efficiently than cholesterol.

Regions of both colicin Ia and diphtheria toxin N-terminal to the channel-forming domains can be translocated across planar phospholipid bilayer membranes. In this article we show that the translocation pathway of diphtheria toxin allows much larger molecules to be translocated than does the translocation pathway of colicin Ia. In particular, the folded A chain of diphtheria toxin is readily translocated by that toxin but is not translocated by colicin Ia. This difference cannot be attributed to specific recognition of the A chain by diphtheria toxin's translocation pathway because the translocation pathway also accommodates folded myoglobin.

The protective antigen component of anthrax toxin forms a homoheptameric pore in the endosomal membrane, creating a narrow passageway for the enzymatic components of the toxin to enter the cytosol. We found that, during conversion of the heptameric precursor to the pore, the seven phenylalanine-427 residues converged within the lumen, generating a radially symmetric heptad of solvent-exposed aromatic rings. This "phi-clamp" structure was required for protein translocation and comprised the major conductance-blocking site for hydrophobic drugs and model cations. We conclude that the phi clamp serves a chaperone-like function, interacting with hydrophobic sequences presented by the protein substrate as it unfolds during translocation.

The 63-kDa fragment of the protective antigen (PA) component of anthrax toxin forms a heptameric channel, (PA63)7, in acidic endosomal membranes that leads to the translocation of edema factor (EF) and lethal factor (LF) to the cytosol. It also forms a channel in planar phospholipid bilayer membranes. What role does this channel play in the translocation of EF and LF? We report that after the 263-residue N-terminal piece of LF (LFN) binds to its receptor on the (PA63)7 channel and its N-terminal end enters the channel at small positive voltages to block it, LFN is translocated through the channel to the opposite side at large positive voltages, thereby unblocking it. Thus, all of the translocation machinery is contained in the (PA63)7 channel, and translocation does not require any cellular proteins. The kinetics of this translocation are S-shaped, voltage-dependent, and occur on a timescale of seconds. We suggest that the translocation process might be explained simply by electrophoresis of unfolded LFN through the channel, but the refolding of the N-terminal half of LFN as it emerges from the channel may also provide energy for moving the rest of the molecule through the channel.

Stable films of didodecyldimethylammonium bromide (DDAB, a synthetic lipid) and horseradish peroxidase (HRP) were made by casting the mixture of the aqueous vesicle of DDAB and HRP onto the glassy carbon (GC) electrode. The direct electron transfer between electrode and HRP immobilized in lipid film has been demonstrated. The lipid films were used to supply a biological environment resembling biomembrane on the surface of the electrode. A pair of redox peaks attributed to the direct redox reaction of HRP were observed in the phosphate buffer solution (pH 5.5). The cathodic peak current increased dramatically while anodic peak decreased by addition of small amount H(2)O(2). The pH effect on amperometric response to H(2)O(2) was studied. The biosensor also exhibited fast response (5 s), good stability and reproducibility.

The electrochemical behavior of horseradish peroxidase (HRP) in the dimyristoyl phosphatidylcholine (DMPC) bilayer on the glassy carbon (GC) electrode was studied by cyclic voltammetry. The direct electron transfer of HRP was observed in the DMPC bilayer. Only a small cathodic peak was observed for HRP on the bare GC electrode. The electron transfer of HRP in the DMPC membrane is facilitated by DMPC membrane. UV-Vis and circular dichroism (CD) spectroscopy were used to study the interaction between HRP and DMPC membrane. On binding to the DMPC membrane the secondary structure of HRP remains unchanged while there is a substantial change in the conformation of the heme active site. Tapping mode atomic force microscopy (AFM) was first applied for the investigation on the structure of HRP adsorbed on supported phospholipid bilayer on the mica and on the bare mica. HRP molecules adsorb and aggregate on the mica without DMPC bilayer. The aggregation indicates an attractive interaction among the adsorbed molecules. The molecules are randomly distributed in the DMPC bilayer. The adsorption of HRP in the DMPC bilayer changes drastically the domains and defects in the DMPC bilayer due to a strong interaction between HRP and DMPC films.

Objective: To study the major histocompatibility complex class II (MHC II) antigen and costimulatory molecules expression on the surface of breast cancer cells., Methods: MHC II antigen and costimulatory molecule expression on five breast cancer cell lines including MCF-7, SK-BR-3, T47D, MDA-MB-435 and ZR-75-30 were detected through flow cytometery analysis, with their expression level compared with that of normal mammary cell line HBL-100., Results: The MHC II expression level of the five breast cancer cell lines were significantly different from that of HBL-100 (P < 0.05). MHC II antigen expression of MCF-7 cells which was about 20 percent of HBL-100 was the lowest. MDA-MB-435 and ZR-75-30 cell expression levels were twice as much as that of HBL-100, with the fluorescence intensity of MDA-MB-435 the highest of all cells. CD40 molecule expression on the surface of MDA-MB-435 cells was the lowest, which was nearly ten percent of that of MCF-7 and HBL-100 cells. CD80 and CD86 molecule expression showed no difference in MDA-MB-435 or HBL-100 cell (P > 0.05), and those of the other four breast cancer cells were lower than that of HBL-100 (P < 0.05)., Conclusion: MHC II antigen and costimulatory molecule expression on the surface of breast cancer cells is abnormal, with different molecule expression in different cells. Breast cancer cells can escape immune surveillance through abnormal molecule expression.

The lipid layer membranes were fabricated on the glassy carbon electrode (GC) and demonstrated to be bilayer lipid membranes by impedance spectroscopy. The formation of incorporated poly L-glutamate bilayer lipid membrane was achieved. The ion channel behavior of the incorporated poly L-glutamate membrane was determined. When the stimulus calcium cations were added into the electrolyte, the ion channel was opened immediately and exhibited distinct channel current. Otherwise, the ion channel was closed. The cyclic voltammogram at the GC electrode coated with incorporated poly L-glutamate DMPC film response to calcium ion is very fast compared with that at the GC electrode coated only with DMPC film. Ion channel current is not dependent on the time but on the concentration of calcium. The mechanism of the ion channel formation was investigated.