Human skin penetration and local effects of topical nano zinc oxide after occlusion and barrier impairment.

Public health concerns continue to exist over the safety of zinc oxide nanoparticles that are commonly used in sunscreen formulations. In this work, we assessed the effects of two conditions which may be encountered in everyday sunscreen use, occlusion and a compromised skin barrier, on the penetration and local toxicity of two topically applied zinc oxide nanoparticle products. Caprylic/capric triglyceride (CCT) suspensions of commercially used zinc oxide nanoparticles, either uncoated or with a silane coating, were applied to intact and barrier impaired skin of volunteers, without and with occlusion for a period of six hours. The exposure time was chosen to simulate normal in-use conditions. Multiphoton tomography with fluorescence lifetime imaging was used to noninvasively assess zinc oxide penetration and cellular metabolic changes that could be indicative of toxicity. We found that zinc oxide nanoparticles did not penetrate into the viable epidermis of intact or barrier impaired skin of volunteers, without or with occlusion. We also observed no apparent toxicity in the viable epidermis below the application sites. These findings were validated by ex vivo human skin studies in which zinc penetration was assessed by multiphoton tomography with fluorescence lifetime imaging as well as Zinpyr-1 staining and toxicity was assessed by MTS assays in zinc oxide treated skin cryosections. In conclusion, applications of zinc oxide nanoparticles under occlusive in-use conditions to volunteers are not associated with any measurable zinc oxide penetration into, or local toxicity in the viable epidermis below the application site.

The need for nano-scale modeling in solid oxide fuel cells.

Solid oxide fuel cells (SOFCs) are high temperature fuel cells, which are being developed for large scale and distributed power systems. SOFCs promise to provide cleaner, more efficient electricity than traditional fossil fuel burning power plants. Research over the last decade has improved the design and materials used in SOFCs to increase their performance and stability for long-term operation; however, there are still challenges for SOFC researchers to overcome before SOFCs can be considered competitive with traditional fossil fuel burning and renewable power systems. In particular degradation due to contaminants in the fuel and oxidant stream is a major challenge facing SOFCs. In this paper we discuss ongoing computational and experimental research into different degradation and design issues in SOFC electrodes. We focus on contaminants in gasified coal which cause electrochemical and structural degradation in the anode, and chromium poisoning which affects the electrochemistry of the cathode. Due to the complex microstructures and multi-physics of SOFCs, multi-scale computational modeling and experimental research is needed to understand the detailed physics behind different degradation mechanisms, the local conditions within the cell which facilitate degradation, and its effects on the overall SOFC performance. We will discuss computational modeling research of SOFCs at the macro-, meso- and nano-scales which is being used to investigate the performance and degradation of SOFCs. We will also discuss the need for a multi-scale modeling framework of SOFCs, and the application of computational and multi-scale modeling to several degradation issues in SOFCs.

Effect of translocation defective reverse transcriptase inhibitors on the activity of N348I, a connection subdomain drug resistant HIV-1 reverse transcriptase mutant.

4'-ethynyl-2-fluoro-2'-deoxyadenosine (EFdA) is a highly potent inhibitor of HIV-1 reverse transcriptase (RT). We have previously shown that its exceptional antiviral activity stems from a unique mechanism of action that is based primarily on blocking translocation of RT; therefore we named EFdA a Translocation Defective RT Inhibitor (TDRTI). The N348I mutation at the connection subdomain (CS) of HIV-1 RT confers clinically significant resistance to both nucleoside (NRTIs) and non-nucleoside RT inhibitors (NNRTIs). In this study we tested EFdA-triphosphate (TP) together with a related compound, ENdA-TP (4'-ethynyl-2-amino-2'-deoxdyadenosine triphosphate) against HIV-1 RTs that carry clinically relevant drug resistance mutations: N348I, D67N/K70R/L210Q/T215F, D67N/K70R/L210Q/T215F/N348I, and A62V/V5I/F77L/F116Y/Q151M. We demonstrate that these enzymes remain susceptible to TDRTIs. Similar to WT RT, the N348I RT is inhibited by EFdA mainly at the point of incorporation through decreased translocation. In addition, the N348I substitution decreases the RNase H cleavage of DNA terminated with EFdA-MP (T/P(EFdA-MP)). Moreover, N348I RT unblocks EFdA-terminated primers with similar efficiency as the WT enzyme, and further enhances EFdA unblocking in the background of AZT-resistance mutations. This study provides biochemical insights into the mechanism of inhibition of N348I RT by TDRTIs and highlights the excellent efficacy of this class of inhibitors against WT and drug-resistant HIV-1 RTs.

Sidewall oxide effects on spin-torque- and magnetic-field-induced reversal characteristics of thin-film nanomagnets.

The successful operation of spin-based data storage devices depends on thermally stable magnetic bits. At the same time, the data-processing speeds required by today's technology necessitate ultrafast switching in storage devices. Achieving both thermal stability and fast switching requires controlling the effective damping in magnetic nanoparticles. By carrying out a surface chemical analysis, we show that through exposure to ambient oxygen during processing, a nanomagnet can develop an antiferromagnetic sidewall oxide layer that has detrimental effects, which include a reduction in the thermal stability at room temperature and anomalously high magnetic damping at low temperatures. The in situ deposition of a thin Al metal layer, oxidized to completion in air, greatly reduces or eliminates these problems. This implies that the effective damping and the thermal stability of a nanomagnet can be tuned, leading to a variety of potential applications in spintronic devices such as spin-torque oscillators and patterned media.

A significant role for Sigma B in the detergent stress response of Listeria monocytogenes.

AIMS: To investigate the contribution of the alternative sigma factor Sigma B to detergent stress in Listeria monocytogenes upon exposure to the surface-active agents, benzalkoniumchloride (BC), cetylpyridinium chloride (CPC) and sodium docecyl sulfate (SDS). METHODS AND RESULTS: Surfactant/detergent stress in L. monocytogenes 10403S and a DeltasigB mutant of 10403S was investigated by lethality, impact on growth and by transcriptional analysis. We observed a significant 1-2 log decrease in the viability of the DeltasigB mutant in response to lethal levels of surfactants. Transcriptional (reverse transcriptase-PCR) analysis revealed the induction of sigmaB by sublethal levels of surface-active agents. However, Sigma B does not play a significant role in the growth of L. monocytogenes upon exposure to sublethal levels as investigated by growth analysis. CONCLUSIONS: Sigma B is essential for the resistance of L. monocytogenes at lethal levels of BC, CPC and SDS. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrate that Sigma B is essential for the resistance of the pathogen to surfactant stress. The findings raise the possibility that induction of Sigma B by sublethal levels of industrial cleaning agents may promote enhanced resistance of further food-processing associated stresses or conditions encountered during infection. Sigma B-regulated mechanisms of detergent resistance may provide targets for the future design of novel cleaning agents.

Use of electrospray ionization mass spectrometry and tandem mass spectrometry to study binding of F0 inhibitors to ceroid lipofuscinosis protein, a model system for subunit c of mitochondrial ATP synthase.

Ceroid lipofuscinosis protein (CLP), the major accumulating protein in several forms of ceroid lipofuscinosis, has an amino acid sequence that is identical to that of the F0 subunit c of normal bovine ATP synthase. Electrospray ionization mass spectrometry (ESI-MS) has shown that ovine CLP and normal bovine F0 subunit c are identical, including a 42 mass unit post-translational modification. Although the identity and the location of this modification have not been fully established in both species, CLP can be used as a convenient and a unique source of subunit c for studies of F0 inhibitor interactions by ESI-MS analysis. Analysis of mixtures of CLP incubated with several known F0 inhibitors showed that N, N'-dicyclohexylcarbodiimide and organotins bind covalently to CLP but interactions with oligomycin and venturicidin were not observed. The sulphydryl inhibitors, 2,3-dimethoxy-5-methyl-1,4,-benzoquinone (UQ0) and N-ethyl maleimide (NEM) were also shown to bind covalently to the protein. The binding stoichiometry and the relative rate of reaction were then determined for each inhibitor. Tandem mass spectrometry experiments performed on the [M+5H]5+ ion of the intact CLP and of the complexes UQ0-CLP and NEM-CLP allowed the identification of 80% of the CLP sequence and revealed that UQ0 and NEM are both bound to cysteine-64. This work shows the exceptional utility of ESI-MS in studies of the interaction of CLP with a range of inhibitors which are applicable to studies of the F0 component of ATP synthase.

Covalent linkage of ruthenium polypyridyl compounds to poly(L-lysine), albumins, and immunoglobulin G.

A series of ruthenium polypyridyl complexes has been covalently bound to poly(L-lysine), bovine serum albumin, human serum albumin, ovalbumin, and immunoglobulin G using different binding methods. The conjugation ratios and the luminescence properties of the bioconjugates are reported. All conjugates show nonsingle-exponential decay curves. Quenching of the emission by oxygen has been studied.

Cytotoxic properties of salivary oxidants.

Salivary peroxidase and to a lesser extent myeloperoxidase are present in significant concentrations in saliva and catalyze the oxidation of thiocyanate anion (SCN-) by H2O2 to yield the potent oxidants hypothiocyanous acid (HOSCN) and its conjugate base hypothiocyanite anion (OSCN-). The objective of this study was to characterize the cytotoxic potential of peroxidase-generated HOSCN/OSCN- toward human erythrocytes. We found that HOSCN/OSCN- (0.25 mM) generated by the peroxidase-H2O2-SCN- system caused significant hemolysis at pH 6.0 but not at pH 6.5, 7.0, or 7.4. Erythrocyte hemoglobin (OxyHb) was oxidized to methemoglobin (MetHb) at all pH values tested; however, the rate of MetHb formation was dramatically increased at low pH and was not affected by inosine hexaphosphate, suggesting that hemoglobin was oxidized primarily by HOSCN. Concurrent with oxidation of hemoglobin (Hb), there was a pH-dependent consumption of HOSCN/OSCN- with more of the oxidant consumed at pH 6.0 compared with pH 6.5, 7.0, or 7.4. The enhanced oxidation of Hb at acidic pH was not due simply to increased membrane permeability by the uncharged species (HOSCN), since both erythrocyte lysate Hb and purified Hb were oxidized to the same extent at low pH as were intact erythrocytes. It is concluded that both OSCN- and HOSCN enter human erythrocytes where the protonated oxidant (HOSCN) mediates hemolysis and oxidizes OxyHb to MetHb, whereas both HOSCN and OSCN- oxidize glutathione (GSH). These data suggest that the extracellular pH may play an important role in modulating the cytotoxic properties of salivary oxidants.

The contractile and control sites of natural actomyosin.

The various contractile and control sites of natural actomyosin gel were studied by comparing the kinetics of ATP hydrolysis with those of gel contraction, measured as an increase in turbidity. Contraction of actomyosin gel seems to require the cooperative reaction of ATP (with Mg) at two different sites. One of these sites catalyzes the hydrolysis of ATP and most probably contributes the driving force for contraction; the binding of ATP to the other site appears to break certain links that retard movement of the gel components. At limiting concentrations of ATP, the rate of contraction seems to depend on the rate of breaking these links as well as on the rate of ATP hydrolysis. But when both sites are saturated, the rate of contraction appears to be limited only by the rate of ATP hydrolysis. In addition to these two contractile sites, there are also two different control sites. At one, the relaxing site, the binding of ATP with Mg inhibits ATP hydrolysis and gel contraction. At the other, the binding of calcium activates contraction by overcoming the inhibitory action of Mg and ATP at the relaxing site. This control system-inhibition by substrate and disinhibition by calcium-can be selectively inactivated by heat and reactivated by dithiothreitol, a disulfide-reducing agent. These observations on the isolated contractile system are discussed in relation to the contraction and relaxation of muscle.

Heat inactivation of the relaxing site of actomyosin: prevention and reversal with dithiothreitol.

Adenosine triphosphate and magnesium (MgATP) inhibit contraction by binding to a specific relaxing site on natural actomyosin gel. This inhibitory control site is distinct from the active sites where MgATP causes contraction. In high concentrations of MgATP, calcium triggers contraction by releasing the protein from substrate inhibition, allowing the contractile reactions to occur. Heating the protein for 5 minutes at 43 degrees C selectively inactivates the relaxing site. After this treatment, actomyosin with MgATP contracts as well without calcium as with it. That this effect of heat is prevented and reversed by dithiothreitol (an agent that reduces disulfide bonds) indicates that the structure of the relaxing site depends on certain labile sulfhydryl groups, which may be those of tropomyosin. When these are oxidized to disulfide bonds, the site loses its activity; when the disulfide bonds are reduced, the site regains its activity.