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Overproduction of melanin can lead to medical disorders such as postinflammatory melanoderma and melasma. Therefore, developing antimelanogenic agents is important for both medical and cosmetic purposes. In this report, we demonstrated for the first time that the antidiabetic drug voglibose is a potent antimelanogenic agent. Voglibose is a representative antidiabetic drug possessing inhibitory activity towards human α-glucosidase; it blocked the proper N-glycan modification of tyrosinase, resulting in a dramatic reduction of the tyrosinase protein level by altering its stability and subsequently decreasing melanin production. Acarbose, another antihyperglycaemic drug that has a lower inhibitory effect on human intracellular α-glucosidase compared with voglibose, did not cause any changes in either the N-glycan modification of tyrosinase or the tyrosinase protein level, indicating that voglibose was the most efficient antimelanogenic agent among the widely used antihyperglycaemic agents. Considering that voglibose was originally selected from the valiolamine derivatives in a screen for an oral antidiabetic drug with a strong inhibitory activity towards intestinal α-glucosidase and low cell permeability, we propose an alternative strategy for screening compounds from valiolamine derivatives that show high inhibitory activity towards human intracellular α-glucosidases and high cell permeability, with the goal of obtaining antimelanogenic agents that are effective inside the cells., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The tremendous interest in magnetic nanoparticles (MNPs) is reflected in published research that ranges from novel methods of synthesis of unique nanoparticle shapes and composite structures to a large number of MNP characterization techniques, and finally to their use in many biomedical and nanotechnology-based applications. The knowledge gained from this vast body of research can be made more useful if we organize the associated results to correlate key magnetic properties with the parameters that influence them. Tuning these properties of MNPs will allow us to tailor nanoparticles for specific applications, thus increasing their effectiveness. The complex magnetic behavior exhibited by MNPs is governed by many factors; these factors can either improve or adversely affect the desired magnetic properties. In this report, we have outlined a matrix of parameters that can be varied to tune the magnetic properties of nanoparticles. For practical utility, this review focuses on the effect of size, shape, composition, and shell-core structure on saturation magnetization, coercivity, blocking temperature, and relaxation time.

Adult skin stem cells are considered an attractive cell resource for therapeutic potential in aged skin. We previously reported that multipotent human dermal stem/progenitor cells (hDSPCs) can be enriched from (normal human dermal fibroblasts (NHDFs) using collagen type IV. However, the beneficial effects of hDSPCs on aged skin remain to be elucidated. In the present study, we analyzed the growth factors secreted from hDSPCs in conditioned medium (CM) derived from hDSPCs (hDSPC-CM) and found that hDSPCs secreted higher levels of bFGF, IGFBP-1, IGFBP-2, HGF, VEGF and IGF-1 compared with non-hDSPCs. We then investigated whether hDSPC-CM has an effect on ultraviolet A (UVA)-irradiated NHDFs. Real-time RT-PCR analysis revealed that the treatment of UVA-irradiated NHDFs with hDSPC-CM significantly antagonized the UVA-induced up-regulation of the MMP1 and the UVA-induced down-regulation of the collagen types I, IV and V and TIMP1 mRNA expressions. Furthermore, a scratch wound healing assay showed that hDSPC-CM enhanced the migratory properties of UVA-irradiated NHDFs. hDSPC-CM also significantly reduced the number of the early and late apoptotic cell population in UVA-irradiated NHDFs. Taken together, these data suggest that hDSPC-CM can exert some beneficial effects on aged skin and may be used as a therapeutic agent to improve skin regeneration and wound healing.

Mutation in the human PTEN-induced protein kinase 1 (PINK1) gene is responsible for the second most common form of recessive Parkinson disease (PD). We have identified a single heterozygous PINK1 mutation, P209A, from a cohort of 68 patients with early onset PD. From age 31, this patient developed an asymmetric bradykinesia with rigidity that was L-DOPA responsive. An [(18)F]-fluorodopa PET scan showed reduced DOPA uptake in the bilateral basal ganglia. The H2O2-induced cell death, ROS production, and caspase-3 activation in SH-SY5Y cells were enhanced by the transfection of the PINK1 P209A mutant. The heme oxygenase-1 (HO-1) induction in response to H2O2 and MPP(+) treatment was impaired by the overexpression of the PINK1 P209A mutant. In addition, SOD2 induction after TNFα treatment was also inhibited by the PINK1 P209A mutation. Akt and ERK are involved in HO-1 induction after oxidative stress. The phosphorylation of Akt and ERK after exposure to H2O2 or MPP(+) was also inhibited in PINK1 P209A mutant cells compared with empty-vector-transfected cells. These results indicate a novel pathway by which the P209A defect in the PINK1 kinase domain inhibits oxidative stress-induced HO-1 and SOD2 induction, which may accelerate the neurodegeneration in PD with PINK1 defect., (Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.)

Adult stem cells from the dermis would be an attractive cell source for therapeutic purposes as well as studying the process of skin aging. Several studies have reported that human dermal stem/progenitor cells (hDSPCs) with multipotent properties exist within the dermis of adult human skin. However, these cells have not been well characterized, because methods for their isolation or enrichment have not yet been optimized. In the present study, we enriched high side scatter (SSC(high))-hDSPCs from normal human dermal fibroblasts using a structural characteristic, intracellular granularity, as a sorting parameter. The SSC(high)-hDSPCs had high in vitro proliferation properties and expressed high levels of SOX2 and S100B, similar to previously identified mouse SOX2+ hair follicle dermal stem cells. The SSC(high)-hDSPCs could differentiate into not only mesodermal cell types, for example, adipocytes, chondrocytes, and osteoblasts, but also neuroectodermal cell types, such as neural cells. In addition, the SSC(high)-hDSPCs exhibited no significant differences in the expression of nestin, vimentin, SNAI2, TWIST1, versican, and CORIN compared with non-hDSPCs. These cells are therefore different from the previously identified multipotent fibroblasts and skin-derived progenitors. In this study, we suggest that hDSPCs can be enriched by using characteristic of their high intracellular granularity, and these SSC(high)-hDSPCs exhibit high in vitro proliferation and differentiation potentials.

This manuscript describes the synthesis of uniform monodisperse SnO2-coated gold nanoparticles and examines their colloidal stability as function of pH, with direct comparison to better known and widely used SiO2-coated gold nanoparticles. Aqueous acidic and basic colloidal SnO2-coated and SiO2-coated Au nanoparticle solutions were prepared, and their stability was monitored visually and by UV-vis spectroscopy. Notably, the SnO2-coated Au nanoparticle solutions were stable up to pH 12.5. However, at pH 13 and 14, the SnO2-coated Au nanoparticles underwent aggregation, which could be fully reversed upon neutralization of the solutions. In contrast, the SiO2-coated Au nanoparticle solutions were unstable at pH>10.5, irreversibly producing a precipitate composed of bare Au nanoparticle aggregates having little or no silica coating. Under acidic conditions, sedimentation was observed from both the colloidal SnO2-coated and SiO2-coated Au nanoparticle solutions, but the colloidal solutions could be reconstituted upon neutralization of the acidic solutions. The sedimentation at low pH coincided with the reported isoelectric pH values of SiO2 and SnO2, respectively. From an applications perspective, we are seeking to develop SnO2-coated metal nanoparticles as stable alternatives to the more widely employed SiO2-coated nanoparticles, with a particular emphasis on their use in sensor devices and solar cells.

By systematically varying the reaction parameters in a liquid-phase reduction reaction, large FeCo nanocubes with tunable body diagonal lengths of 175, 350, and 450 nm were synthesized. The nanocubes were initially stabilized with poly(vinyl pyrrolidone) (PVP) and then coated with a relatively thin layer of silica (~55 nm thick), which allowed them to retain their cubic shape. The magnetization curves showed that the PVP-stabilized nanocubes exhibited a high saturation magnetization of 167 ± 4 emu/g. The saturation magnetization, however, decreased upon coating with silica to 146 ± 13 emu/g for the particles with 350 and 450 nm FeCo cores and 48 ± 1 emu/g for the particles with 175 nm FeCo cores. The silica-coated FeCo nanocubes were then functionalized with 3-(aminopropyl)-trimethoxysilane (APTMS), and a layer of surface-bound nanoparticle was generated by exposing the resultant amine-functionalized nanocubes to self-assembled monolayers (SAMs) on gold terminated with carboxylic-acid groups.

Cancer cells evade replicative senescence by re-expressing telomerase, which maintains telomere length and hence chromosomal integrity. Telomerase inhibition would lead cancer cells to senesce and therefore prevent cancer cells from growing indefinitely. G-quadruplex ligands can attenuate telomerase activity by inducing G-quadruplex formation at the 3'-overhang of telomere and at the human telomerase reverse transcriptase (hTERT) promoter; the former prevents telomerase from accessing the telomere, and the latter acts as a transcriptional silencer. The present investigation found that perylene derivatives PM2 and PIPER induced G-quadruplex formation from both telomeric DNA and the hTERT promoter region in vitro. Further, TRAP assay showed that these compounds inhibited telomerase in a dose-dependent manner. When A549 human lung cancer cells were treated with these compounds, hTERT expression was down-regulated. Moreover, the crude protein extract from these treated cells exhibited less telomerase activity. In the long-term treatment of A549 lung cancer cells with sub-cytotoxic dose of these perylenes, telomere shortening, reduction of cell proliferation and tumorigenicity, and cell senescence were observed. The results of this study indicate that perylene derivatives warrant further consideration as effective agents for cancer therapy., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Skin diseases can be characterized by their predominant CD4-positive T-helper (Th) cell profiles. Chronic dermatological conditions often give rise to abnormal skin pigmentation. To understand the role of Th cells in pigmentation, the effects of the major Th cell cytokines, IFNγ, IL-4, and IL-17A, on melanogenesis were evaluated using cultured normal human melanocytes (NHMs) instead of relying on transformed melanoma cell lines. IL-4 directly inhibited melanogenesis in NHMs and downregulated both transcription and translation of melanogenesis-associated genes, such as microphthalmia-associated transcription factor (MITF) and dopachrome tautomerase. Despite the lack of a direct inhibition of melanin pigment synthesis, IFNγ and IL-17A increased the synthesis of an antimelanogenic cytokine IL-6 in NHMs. IFNγ activated signal transducers and activators of transcription 1 (STAT1) and STAT3 phosphorylation in NHMs, and IL-4 increased the STAT3 and STAT6 phosphorylation. The differential phosphorylation profile of STAT proteins between IFNγ and IL-4 may explain the difference in their effect on melanogenesis in NHMs. The IL-4-induced downregulation of melanogenesis was inhibited by treating NHMs with a JAK2 inhibitor AG490 or STAT6 siRNA. In conclusion, the involvement of the IL-4-induced JAK2-STAT6 signaling and the IFNγ- or IL-17A-dependent antimelanogenic IL-6 production should be considered as one of the mechanisms explaining the association with hypopigmention in skin diseases.

Haptens must react with cellular proteins to be recognized by antigen presenting cells. Therefore, monitoring reactivity of chemicals with peptide/protein has been considered an in vitro skin sensitization testing method. The reactivity of peptides with chemicals (peptide reactivity) has usually been monitored by chromatographic methods like HPLC or LC/MS, which are robust tools for monitoring common chemical reactions but are rather expensive and time consuming. Here, we examined the possibility of using spectrophotometric methods to monitor peptide reactivity. Two synthetic peptides, Ac-RWAACAA and Ac-RWAAKAA, were reacted with 48 chemicals (34 sensitizers and 14 non-sensitizers). Peptide reactivity was measured by monitoring unreacted peptides with UV-Vis spectrophotometer using 5,5'-dithiobis-2-nitrobenzoic acid as a detection reagent for the free thiol group of cysteine-containing peptide or fluorometer using fluorescamine™ as a detection reagent for the free amine group of lysine-containing peptide. Chemicals were categorized as sensitizers when they induced more than 10% depletion of cysteine-containing peptide or 20% depletion of lysine-containing peptide. The sensitivity, specificity, and accuracy of this method were 82.4%, 85.7%, and 83.3%, respectively. These results demonstrate that spectrophotometric methods can be easy, fast, and high-throughput screening tools for the prediction of the skin sensitization potential of chemical haptens., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The tridentate adsorbates cyclohexane-cis,cis-1,3,5-triyltrimethanethiol (CyTSH) and (cis,cis-1,3,5-trimethylcyclohexane-1,3,5-triyl)trimethanethiol (3C1CyTSH) were designed and synthesized. Thin films prepared by the adsorption of these molecules onto the surface of gold were characterized by ellipsometry, contact angle goniometry, polarization modulation infrared reflection absorption spectroscopy, and X-ray photoelectron spectroscopy (XPS). CyTSH was found to generate multilayer thin films with many unbound thiol species and oxidized sulfur moieties (e.g., disulfides and sulfones). In contrast, 3C1CyTSH was found to generate monolayer films in which ~90% of the thiols were bound to the surface of gold (~10% unbound), and there were no oxidized sulfur species. In comparing CyTSH and 3C1CyTSH, the methyl groups of 3C1CyTSH impart rigidity to the structure, which significantly enhances the chemisorption of sulfur to the surface of gold. Ellipsometric measurements and analysis by XPS indicate that the thickness of the self-assembled monolayer formed from 3C1CyTSH is ~5 Å.

Although most nanofabrication techniques can control nano/micro particle (NMP) size over a wide range, the majority of NMPs for biomedical applications exhibits a diameter of ~100 nm. Here, the vascular distribution of spherical particles, from 10 to 1,000 nm in diameter, is studied using intravital microscopy and computational modeling. Small NMPs (≤100 nm) are observed to move with Red Blood Cells (RBCs), presenting an uniform radial distribution and limited near-wall accumulation. Larger NMPs tend to preferentially accumulate next to the vessel walls, in a size-dependent manner (~70% for 1,000 nm NMPs). RBC-NMP geometrical interference only is responsible for this behavior. In a capillary flow, the effective radial dispersion coefficient of 1,000 nm particles is ~3-fold larger than Brownian diffusion. This suggests that sub-micron particles could deposit within diseased vascular districts more efficiently than conventional nanoparticles.

Melanoblasts are derived from the neural crest and migrate to the dermal/epidermal border of skin and hair bulges. Although melanoblast migration during embryogenesis has been well investigated, there are only a few reports regarding the migration of mature melanocytes. Here, we demonstrate that a chemokine, stromal-derived factor-1 (SDF1, also known as CXCL12), and one of its receptor CXCR7 regulate normal human epidermal melanocyte (NHEM) migration. We found that SDF1 induces the directional migration of NHEMs. Interestingly, although both CXCR4 and CXCR7 are expressed in NHEMs, blockade of CXCR4 using a CXCR4-specific neutralizing antibody did not exert any influence on the SDF1-induced migration of NHEMs, whereas blockade of CXCR7 using a CXCR7-specific neutralizing antibody did influence migration. Furthermore, SDF1-induced NHEMs migration exhibited the early hallmark events of CXCR7 signaling associated with MAP kinase activation. It is known that the phosphorylation of ERK through CXCR7 signaling is mediated by β-arrestins. The treatment of NHEMs with SDF1 resulted in the phosphorylation of ERK in a β-arrestin 2-dependent manner. These results suggest that melanocytes may have a unique mechanism of migration via SDF1/CXCR7 signaling that is different from that of other cell types., (© 2012 John Wiley & Sons A/S.)

Respiratory morbidity (particularly COPD and asthma) can be influenced by short-term weather fluctuations that affect air quality and lung function. We developed a model to evaluate meteorological conditions associated with respiratory hospital admissions in the Shenandoah Valley of Virginia, USA. We generated ensembles of classification trees based on six years of respiratory-related hospital admissions (64,620 cases) and a suite of 83 potential environmental predictor variables. As our goal was to identify short-term weather linkages to high admission periods, the dependent variable was formulated as a binary classification of five-day moving average respiratory admission departures from the seasonal mean value. Accounting for seasonality removed the long-term apparent inverse relationship between temperature and admissions. We generated eight total models specific to the northern and southern portions of the valley for each season. All eight models demonstrate predictive skill (mean odds ratio = 3.635) when evaluated using a randomization procedure. The predictor variables selected by the ensembling algorithm vary across models, and both meteorological and air quality variables are included. In general, the models indicate complex linkages between respiratory health and environmental conditions that may be difficult to identify using more traditional approaches.

The expression of Wnt inhibitory factor-1 (WIF-1) gene, which was detected by a microarray analysis of hyperpigmented and normally pigmented skin sets of melasma patients, was significantly reduced in the hyperpigmented skin from melasma patients, but not in healthy controls, regardless of UV irradiation. Wnt signals regulate skin pigmentation; however, WIF-1 is expressed in cultured skin keratinocytes and fibroblasts, but not in melanocytes. Therefore, we examined whether WIF-1 knockdown in neighboring keratinocytes and fibroblasts plays a role in melasma. Additionally, the effect of WIF-1 overexpression on the amelioration of hyperpigmentation was examined. WIF-1 knockdown, either in fibroblasts or in keratinocytes, significantly stimulated tyrosinase expression and melanosome transfer, whereas melanocytes with WIF-1 overexpression significantly reduced those parameters. The WIF-1 knockdown decreased glycogen synthase kinase-3β (GSK-3β), β-catenin, and NFATc2 (nuclear factor of activated T cells, cytoplasmic, calcineurin-dependent 2) phosphorylation and increased microphthalmia-associated transcription factor (MITF) expression as in melanocytes with Wnt-1 overexpression, whereas the WIF-1 overexpression reversed the results. Expression of Wnts, both canonical and noncanonical, was increased in the hyperpigmented skin of melasma patients. Collectively, WIF-1 downregulation, which may occur in epidermal keratinocytes and in dermal fibroblasts, is involved in melasma development because of the stimulation of melanogenesis and melanosome transfer through upregulation of the canonical and the noncanonical Wnt signaling pathway.

Self-assembled monolayers (SAMs) were prepared from a novel two-tailed partially fluorinated thiol (F8C11/C16), possessing one hydrocarbon chain and one chain with an extended fluorinated segment, and from mixtures of F8C11/C16 and hexadecanethiol (C16) on gold, with the expectation that the internal chemical dissimilarity and wedge-like shape of F8C11/C16 would lead to unique self-organizational motifs. The SAMs were systematically characterized using ellipsometry, atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), contact angle goniometry, and polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS). Based on this characterization, the one-component F8C11/C16 SAMs exhibited relatively poor molecular organization compared to traditional alkanethiols, forming low coverage monolayers with significant molecular disorder. However, the series of mixed SAMs formed from F8C11 and F8C11/C16 were anomalously well ordered as indicated by film thickness, surface coverage, and the frequencies of characteristic vibrational modes. AFM images of these mixed SAMs exhibited nanoscale fibrillar structures in a birds-nest morphology, suggesting that in the presence of a C16 matrix, the F8C11/C16 component organized into the two-dimensional analogue of discrete bilayers. Control experiments involving mixed SAMs comprised of F8C11/C16 and a single-tailed partially fluorinated thiol (F8C11) or C16 and F8C11 exhibited no appreciable indication of interesting self-organization beyond an evenly dispersed mixing of the thiolates or phase separation, respectively.

Custom-designed surfactants, known as "linactants", have the ability to reduce the line tension between coexisting phases within mixed monolayers of chemically dissimilar compounds at the air-water interface. Thus far, linactants have been successfully identified for only one type of chemical dissimilarity, involving mixed monolayers of hydrocarbon and fluorocarbon surfactants. In the present work, we have pursued a more general interpretation of linactant compounds by extending the concept to a new system that is comprised of a mixture of aliphatic (pentadecanoic acid) and aromatic (p-terphenyl carboxylic acid) compounds. We found that the "bare" line tension between phases of this mixed monolayer was ~4 pN, and within the same order of magnitude as our previous measurement in mixed monolayers containing hydrocarbons and fluorocarbons. Furthermore, we examined a homologous series of potential linactant compounds possessing an aliphatic tail of variable length and a p-terphenyl block. We determined that linactants with longer tails were able to reduce the line tension more efficiently and effectively. In particular, the addition of only 0.14% of a linactant with an 11-carbon chain reduced the line tension by more than a factor of 2. We hypothesize that the efficiency of this particular linactant is associated with its long tail; this creates strong van der Waals interactions with the aliphatic chains and enables the tail to adopt conformations that facilitate π-stacking interactions with the aromatic compounds within the monolayer.

Aims: The activation of Wnt/β-catenin signaling pathway plays an important role in hair follicle morphogenesis by stimulating bulge stem cells. This study was to obtain the activator of Wnt/β-catenin signaling pathway from natural products and to determine whether this activator can induce anagen hair growth in mice., Main Methods: To identify materials that activate Wnt/β-catenin signaling pathway, 800 natural product extracts were screened using pTOPFlash assay and neural progenitor cell (NPC) differentiation assay. A selected extract was further tested for its effects on alkaline phosphatase (ALP) activity in human immortalized dermal papilla cell (iDPC) and the proliferation in iDPC and immortalized rat vibrissa DPC (RvDP). Finally, hair growth-promoting effects were evaluated in the dorsal skin of C57BL/6 mice., Key Findings: Aconiti Ciliare Tuber (ACT) extract was one of the most active materials in both pTOPFlash and NPC differentiation assays. It promoted the differentiation of NPC cells even under proliferation-stimulating conditions (basic fibroblast growth factor: bFGF). It also increased ALP activity and proliferation of iDPC in dose-dependent manners, and it stimulated the induction of the anagen hair growth in C57BL/6 mice. These results suggest that ACT extract activates the Wnt/β-catenin signaling pathway by enhancing β-catenin transcription and has the potential to promote the induction of hair growth via activation of the stem cell activity of the dermal papilla cells., Significance: This is the first report indicating benefits of ACT extract in hair loss prevention by triggering the activation of Wnt/β-catenin signaling pathway and induction of the anagen hair growth in mice., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Human skin hyperpigmentation disorders occur when the synthesis and/or distribution of melanin increases. The distribution of melanin in the skin is achieved by melanosome transport and transfer. The transport of melanosomes, the organelles where melanin is made, in a melanocyte precedes the transfer of the melanosomes to a keratinocyte. Therefore, hyperpigmentation can be regulated by decreasing melanosome transport. In this study, we found that an extract of Saururus chinensis Baill (ESCB) and one of its components, manassantin B, inhibited melanosome transport in Melan-a melanocytes and normal human melanocytes (NHMs). Manassantin B disturbed melanosome transport by disrupting the interaction between melanophilin and myosin Va. Manassantin B is neither a direct nor an indirect inhibitor of tyrosinase. The total melanin content was not reduced when melanosome transport was inhibited in a Melan-a melanocyte monoculture by manassantin B. Manassantin B decreased melanin content only when Melan-a melanocytes were co-cultured with SP-1 keratinocytes or stimulated by α-MSH. Therefore, we propose that specific inhibitors of melanosome transport, such as manassantin B, are potential candidate or lead compounds for the development of agents to treat undesirable hyperpigmentation of the skin., (© 2012 John Wiley & Sons A/S.)

The pathogenesis of melasma is unknown, although the potential role of estrogen has been considered. Microarray and real-time PCR analyses revealed that upregulation of PDZ domain protein kidney 1 (PDZK1) is clinically correlated with melasma. Although there has been no report that PDZK1 is involved in pigmentation and/or melanogenesis, PDZK1 expression can be induced by estrogen. In this study, the role of PDZK1 upregulation in melasma was examined, particularly in connection with estrogen, using biopsied skin specimens from 15 patients and monocultures and cocultures of melanocytes and keratinocytes with or without overexpression or knockdown of PDZK1. Estrogen upregulated PDZK1. Overexpression of PDZK1 increased tyrosinase expression and melanosome transfer to keratinocytes, whereas PDZK1 knockdown reduced estrogen-induced tyrosinase expression, through regulation of expression of estrogen receptors (ERs) ER-α and ER-β. The PDZK1-induced tyrosinase expression and melanosome transfer was regulated by ion transporters such as sodium-hydrogen exchanger (NHE), cystic fibrosis transmembrane conductance regulator (CFTR), and SLC26A3, which showed a specific association with each ER subtype. In the melanosome transfer, PDZK1 also increased phosphorylation of ezrin/radixin/moesin (ERM) and ras-related C3 botulinum toxin substrate 1, but not the expression of proteinase-activated receptor-2. Collectively, upregulation of PDZK1 could have an important role in the development of melasma in connection with estrogen through NHE, CFTR, and SLC26A3.

Using a human corneal cell line (HCE-T cells) and 2 evaluation criteria, we developed a new alternative method to assess the eye irritation potential of chemicals. We exposed HCE-T cells to different concentrations of 38 chemicals for 1h and measured relative cell viability (RCV) as an endpoint at each concentration. Using the RCV values, we calculated the RCV50. We also exposed HCE-T cells to 3 fixed concentrations of the 38 chemicals (5%, 0.5%, and 0.05%) for 1h and measured the RCV at each concentration. Using the RCV values at 5%, 0.5%, and 0.05%, we developed a new criterion for eye irritation potential (total eye irritation score, TEIS) and estimated the ocular irritancy. We then assessed the correlation of the results of RCV50 and TEIS with those of the Draize rabbit eye irritation. Both the RCV50 and TEIS results exhibited good positive correlations (sensitivity: 80.77%, specificity: 83.33%, and accuracy: 81.58% for TEIS; sensitivity: 73.08-76.92%, specificity: 75.00%, and accuracy: 73.68-76.32% for RCV50). We conclude that the new in vitro model using HCE-T cells is a good alternative evaluation model for the prediction of the eye irritation potential of chemicals., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Introduction: Shoulder-pelvic integration could play a central role in the control of dynamic posture and movement. However, kinematic coordination during axial trunk rotation has not been carefully investigated in subjects with recurrent low back pain (LBP). The purpose of this study was to compare the maximum rotational angles of the shoulders and pelvis in the transverse plane between subjects with and without recurrent LBP., Materials and Methods: A total of 38 age-matched subjects (19 control subjects: 69.00 ± 5.75 years old and 19 subjects with LBP: 68.79 ± 5.40 years old) participated in the study. The axial trunk rotation test was conducted in the upright position with bilateral hips and knees fully extended and both feet shoulder width apart., Results: The results of this study indicated that there was a difference in pelvic girdle rotation between groups (100.79 ± 26.46 in the control group, 82.12 ± 23.16 in the LBP group; t = 2.31, p = 0.02); however, there was no difference for the shoulder girdle (177.63 ± 36.98 in the control group, 156.42 ± 30.09 in the LBP group; t = 1.91, p = 0.06). There were interactions with age (F = 9.27, p = 0.004) and BMI (F = 7.50, p = 0.01) with the rotational angles of the shoulder and pelvis., Conclusion: These results indicated a different pattern of trunk rotation movement with the age and BMI serving as important factors to consider for recurrent LBP. The results of our study also indicated a different pattern of shoulder and pelvic coordination with age and gender. Clinicians need to consider the consequences of limited shoulder-pelvic rotational angles, especially limited rotational angle on the pelvis during trunk axial rotation. Further studies are required to determine the causes of the underlying problems for clinical decision-making and altered shoulder-pelvic rotation in subjects with recurrent LBP.

Background: Kojic acid is a fungal metabolite widely used in medicinal and cosmetic formulations as a skin-lightening agent based on its de-pigmenting activity. Although in human clinical studies kojic acid has been shown to be effective in the treatment of hyper-pigmentation disorders such as melasma, the reasons for its apparent lack of anti-melanogenic activity in cultured mammalian melanocytes are unclear., Objectives: This study was aimed to elucidate pharmacological mechanisms of the in vivo anti-melanogenic activity of kojic acid in human skin., Methods: A primary human melanocyte and keratinocyte co-culture system was used to evaluate whether kojic-acid-induced changes in keratinocytes were associated with anti-melanogenic activities in melanocytes. The cytokine secretion profiles in response to kojic acid were analyzed., Results: Kojic acid increased interleukin (IL)-6 and IL-8 production in melanocyte/keratinocyte co-cultures; however, IL-6 directly inhibited melanogenesis whereas IL-8 did not. In melanocyte monocultures, kojic acid did not increase IL-6 production whereas in keratinocyte monocultures it significantly up-regulated IL-6 gene and protein expression. Therefore, the up-regulation of IL-6 in melanocyte/keratinocyte co-cultures seems to be originated from kojic acid-induced changes in keratinocytes. Anti-IL-6 antibody treatment antagonized the anti-melanogenic effect of kojic acid on the co-cultures., Conclusions: The pharmacological mechanism of kojic acid to explain clinically effective anti-melanogenic activity on hyper-pigmented skin is associated with the kojic acid-induced IL-6 production in keratinocytes. The cross-talk between melanocytes and keratinocytes should be determined in future studies on the pharmacological mechanisms of clinically effective dermatological drugs acting on the epidermis., (Copyright © 2012 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)

Drosophila melanogaster has been used as a genetic model organism to understand the fundamental molecular mechanisms in human biology including memory formation that has been reported involving protein synthesis and/or post-translational modification. In this study, we employed a proteomic platform based on fluorescent 2DE and MALDI-TOF MS to build a standard D. melanogaster head proteome map for proteome-proteome comparison. In order to facilitate the comparison, an interactive database has been constructed for systematically integrating and analyzing the proteomes from different conditions and further implicated to study human diseases related to D. melanogaster model. In summary, the fundamental head proteomic database and bioinformatic analysis will be useful for further elucidating the biological mechanisms such as memory formation and neurodegenerative diseases., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

IBMPFD, Inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia, is a hereditary degenerative disorder due to single missense mutations in VCP (Valosin-Containing Protein). The mechanisms of how mutations of VCP lead to IBMPFD remain mysterious. Here we utilize two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry to study the IBMPFD disorder at the protein level. With this set-up, we are able to employ comparative proteomics to analyze IBMPFD disease using Drosophila melanogaster as our disease model organism. Head proteome of transgenic D. melanogaster expressing wild type VCP is compared, respectively, with the head proteome of transgenic mutant type VCPs that correspond to human IBMPFD disease alleles (TER94(A229E), TER94(R188Q), and TER94(R152H)). Of all the proteins identified, a significant fraction of proteins altered in TER94(A229E) and TER94(R188Q) mutants belong to the same functional categories, i.e. apoptosis and metabolism. Among these, Drosophila transferrin is observed to be significantly up-regulated in mutant flies expressing TER94(A229E). A knock-down experiment suggests that fly transferrin might be a potential modifier in IBMPFD disease. The molecular analysis of IBMPFD disease may benefit from the proteomics approach which combines the advantages of high throughput analysis and the focus on protein levels.

Background: Ultraviolet (UV) A irradiation causes the degeneration of extracellular matrix in the skin dermis, mainly due to disrupted collagen homeostasis, resulting in the photo-aging of human skin. All-trans retinoic acid (ATRA) improves photo-aged human skin in vivo., Objectives: Although the effects of ATRA on collagen synthesis and MMP regulation are well known, the effects of ATRA on other collagen homeostasis-associated genes have not been elucidated. This study was aimed to study the factors that are pharmacologically associated with the effect of ATRA on collagen homeostasis., Methods: The gene transcription profile of collagen homeostasis-associated genes was systematically evaluated in three-dimensional human dermal equivalents (HDEs) following UVA-irradiation and/or ATRA treatment., Results: In addition to the expected changes in MMPs and collagen synthesis in HDEs in response to ATRA, prolidase, an important enzyme in the recycling of proline and hydroxyproline from degraded collagen molecules, was significantly decreased by UVA irradiation, and its down-regulation was antagonized by ATRA. Transfection with a prolidase-specific siRNA led to a significant decrease in procollagen synthesis in human fibroblasts. ATRA inhibited the UVA irradiation-induced decrease in prolidase activity through an insulin-like growth factor (IGF) receptor signaling pathway in HDEs. ARTA increased IGF1 and IGF2 production in HDEs, and neutralizing IGFs with anti-IGF antibodies abolished the effect of ATRA on proliase activity., Conclusions: These data demonstrate that ATRA regulates prolidase activity in HDEs via IGF receptor signaling, suggesting one of the pharmacological mechanisms by which improves photo-aged human skin., (Copyright © 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)

Background: Cathepsin G, a serine protease that is activated by ultraviolet (UV) radiation, increases matrix metalloproteinase-1 (MMP-1) expression in fibroblasts through fibronectin (Fn) fragmentation and promotes the conversion of proMMP-1 to active MMP-1., Objectives: This study investigated whether [2-[3-[[(1-benzoyl-4-piperidinyl)methylamino]carbonyl]-2-naphthalenyl]-1-(1-naphthalenyl)-2-oxoethyl]-phosphonic acid (KPA), a cathepsin G inhibitor, plays any role in extracellular matrix (ECM) damage in an in vitro 3D dermal equivalent (DE) and an in vivo ultraviolet B (UVB)-irradiated hairless mice., Methods: We examined the potential ECM-protective effects of a cathepsin G inhibitor in an in vitro 3D DE model and an in vivo UVB-irradiated hairless mouse skin model., Results: Among five known serine protease inhibitors, KPA showed the strongest potency and selectivity against cathepsin G. KPA inhibited the cathepsin G-mediated MMP-1 increase and alleviated the downregulation of mRNAs encoding collagen and tissue inhibitor of matrix metalloproteinase-1 in an in vitro 3D DE model. Most importantly, topical application of KPA (0.025%) to the dorsal skin of hairless mice enhanced collagen expression and attenuated UVB-induced Fn fragmentation and upregulation of MMP-2 and MMP-9 activities., Conclusions: Cathepsin G inhibitors may be useful for the prevention of UVB-induced photoaging through amelioration of ECM damage and MMP upregulation., (Copyright © 2012 S. Karger AG, Basel.)

Several studies have reported that 1,2-alkanediols show increasing anti-microbial activity as their alkane chain length increases. However, there are no reports on the influence of alkane chain length on the skin irritation potential of 1,2-alkanediols. To investigate the influence of alkane chain length on the skin irritation potential of 1,2-alkanediols. The objective and subjective (sensory) skin irritation potentials of five 1,2-alkanediols - 1,2-butanediol, 1,2-pentanediol, 1,2-hexanediol, 1,2-octanediol and 1,2-decanediol - were evaluated. We also estimated percutaneous absorption by measuring in vitro skin penetration using a Franz diffusion cell system. Like anti-microbial activity, sensory irritation potential increased as alkane chain length increased, most likely due to increasing membrane interference and/or intrinsic toxicity of 1,2-alkanediols. 1,2-Hexanediol showed the lowest objective skin irritation potential, which increased when the alkane chain length decreased or increased. Furthermore, percutaneous absorption negatively correlated with the alkane chain length of 1,2-alkanediols. These results show that a lower skin absorption potential is not indicative of a low skin irritation potential. Our results suggest that the factors and processes involved in skin irritation potential are complex and that skin irritation potential is influenced by intrinsic toxicity and the potential for penetration or integration in the lipid bilayer., (© 2011 The Authors. ICS © 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie.)

Hollow gold-silver nanoshells having systematically varying sizes between 40 and 100 nm were prepared. These particles consist of a hollow spherical silver shell surrounded by a thin gold layer. By varying the volume of the gold stock solution added to suspensions of small silver-core templates, we tailored the hollow gold-silver nanoshells to possess strong tunable optical extinctions that range from the visible to the near-IR spectral regions, with extinctions routinely centered at ∼950 nm. The size and morphology of these core/shell nanoparticles were characterized by dynamic light scattering (DLS), field emission scanning electron microscopy (FE-SEM), and transmission electron microscopy (TEM). Separately, X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS) were used for measuring their elemental composition; UV-vis spectroscopy was used to evaluate their optical properties. Given their relatively small size compared to other nanoparticles that absorb strongly at near IR wavelengths, these easy-to-synthesize particles should find use in applications that require ultrasmall nanoparticles with extinctions comfortably beyond visible wavelengths (e.g., medicinal therapies, diagnostic imaging, nanofluidics, and display technologies).

This article describes the design, synthesis, and study of alkoxyphenylethanethiol-based adsorbates with one (R1ArMT), two (R2ArMT), and three (R3ArMT) pendant octadecyloxy chains substituted at the 4-, 3,5-, and 3,4,5-positions, respectively, of the phenylethanethiol group. These adsorbates are being developed for use in the preparation of compositionally versatile "mixed" self-assembled monolayer (SAM) coatings. The resultant SAMs were characterized by ellipsometry, contact angle goniometry, polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS), and X-ray photoelectron spectroscopy (XPS). The studies revealed that R1ArMT generates a well-ordered monolayer film, while R2ArMT and R3ArMT generate monolayer films with diminished conformational order in which the degree of crystallinity decreases as follows: C18 ∼ R1ArMT > R3ArMT > R2ArMT. In addition, comparison of the molecular and chain packing densities of SAMs derived from these new adsorbates reveals that the R2ArMT and R3ArMT adsorbates give rise to SAMs with reduced chain tilt and smaller surface area per chain when compared to the SAMs derived from C18 and R1ArMT.

Background: The aim of this study was to develop a method for the evaluation of subtle change in skin roughness caused by cleansing products under mild application conditions using a non-invasive three-dimensional (3D) analysis system., Methods: A double-blind comparative study of the modified soap chamber test was performed using two soap bars and a syndet bar. Skin changes were evaluated by visual scoring [mean cumulative irritation index (MCII)] and by bioengineering measurements [transepidermal water loss (TEWL), skin capacitance, and skin surface roughness]., Results: MCII of the syndet bar was statistically higher than that of one soap bar, and TEWL increase after application of the syndet bar was statistically higher than that of both soap bars. Skin capacitance decreased significantly only after application of the syndet bar. The change in the average roughness of the skin surface was significantly greater after the application of the syndet bar than with classic soap bars., Conclusion: A simple, fast, and objective evaluation of skin surface topography was performed using a modified soap chamber test and a non-invasive 3D analysis system. The results suggest that measurement of skin roughness using a non-invasive 3D analysis system might be a good method for the evaluation of a subtle change caused by cleansing products under mild application conditions., (© 2011 John Wiley & Sons A/S.)

The spontaneous adsorption of organic molecules on a variety of planar and nonplanar substrates, that is, self assembly, can generate films just one molecule thick. These nanoscale, self-assembled monolayer (SAM) films have been extensively used to engineer surfaces with well-defined properties. Their utility has been demonstrated in a wide range of applications, including wetting, adhesion, lubrication, patterning, and molecular recognition. Many SAM systems have been investigated, but alkanethiols adsorbed on gold are the most successful combination. This pairing offers a variety of advantages, including the ability to tune precisely the interfacial properties of a surface through the well-established organic synthetic methodologies that have been developed for preparing custom ω-terminated alkanethiols. Alkanethiolate monolayers are moderately stable at room temperature; however, these films degrade over time and readily desorb upon moderate heating. This shortcoming limits the use of SAMs in applications involving elevated temperatures or harsh environments. Accordingly, new adsorbates with multiple bonding moieties have been created to enhance the stability and versatility of SAMs. In this Account, we examine a variety of multidentate adsorbate structures that have been used to generate SAMs on planar substrates and on nanoparticles. Each of these chelating adsorbates (bidentates and tridentates) has been designed to generate well-defined organic monolayer films with multiple attachment points to the underlying substrate. This bonding arrangement allows the formation of SAMs with enhanced stability through the entropy-driven "chelate effect". The research examined here demonstrates that multidentate adsorbates provide robust films: they enable the use of SAMs under conditions that are incompatible with SAMs derived from normal alkanethiols. Another advantage offered by multidentate adsorbates is the capacity for new paradigms in thin-film composition. In particular, appropriately designed chelating adsorbates can be engineered to have two or more chemically distinct terminal groups that are covalently linked to the same underlying headgroup, without adding steric bulk that might prove detrimental to the resultant assembly. This strategy allows the generation of homogeneously mixed multicomponent surfaces, overcoming the problem of phase separation or "islanding" that is pervasive when two or more chemically distinct adsorbates are used to form mixed SAMs. Such homogeneously mixed films offer the opportunity to fine-tune the interfacial properties of a substrate and to create unique heterogeneous interfaces that are well defined by the chemical composition of the tailgroups exposed at the surface. The insight derived from these studies opens the door to new uses for SAMs, both in surface engineering applications (such as corrosion resistance and soft lithographic patterning) and in the stabilization and manipulation of nanoparticles.

Parkinson's disease (PD) is one of the most common neurodegenerative diseases. Mutation in the phosphatase and tensin homolog (PTEN)-induced putative kinase 1 (PINK1) gene causes an autosomal recessive form of PD. However, the etiology related to PINK1 is still not clear. Here, we examined the effect of PINK1 on heme oxygenase (HO)-1 induction in SH-SY5Y neuronal cells following H(2)O(2) or 1-methyl-4-phenylpyridinium (MPP(+)) treatment. The HO-1 induction in response to H(2)O(2) and MPP(+) treatment was impaired by the expression of recombinant PINK1 G309D mutant. PINK1 G309D mutation increased the apoptosis of SH-SY5Y cells following H(2)O(2) treatment and cell survival was rescued by the over-expression of HO-1 using adenovirus (Ad) infection. In addition, knockdown of tumor necrosis factor receptor-associated protein-1 (TRAP1), which is the substrate of PINK1 kinase, in SH-SY5Y cells also inhibited the expression of HO-1 in response to oxidative stress. The up-regulation of TRAP1 expression following H(2)O(2) treatment was inhibited by the expression of recombinant PINK1 G309D mutant. The H(2)O(2)-induced HO-1 induction was Akt- and ERK-dependent. The phosphorylation of ERK and Akt but not p38 was inhibited in cells expressing the PINK1 G309D mutant and knockdown of TRAP1. These results indicate a novel pathway by which the defect of PINK1 inhibits the oxidative stress-induced HO-1 production. Impairment of HO-1 production following oxidative stress may accelerate the dopaminergic neurodegeneration in Parkinson patients with PINK1 defect., (© 2011 The Authors. Journal of Neurochemistry © 2011 International Society for Neurochemistry.)

Microfluidic channels have received much attention because they can be used to control and transport nanoscale objects such as nanoparticles, nanowires, carbon nanotubes, DNA and cells. However, so far, practical channels have not been easy to design because they require very expensive fabrication and sensitive experiments. Numerical approaches can be alternatives or supplementary measures to predict the performance of new channels because they efficiently explain nanoscale multi-physics phenomena and successfully solve nanowire alignment and cell adhesion problems. In this paper, a newly updated immersed finite element method that accounts for collision force and Brownian motion as well as fluid-solid interaction is proposed, and is applied to simulate nanoparticle movements in a microfluidic channel. As part of the simulation, Brownian motion effects in a single nanoparticle focusing lens system are examined under different temperature conditions, and the resulting transport efficiencies are discussed. Furthermore, nanoparticle movements in a double focusing lens system are predicted to show the enhancement of focusing efficiency.

Background: Over-expression of Aurora kinases promotes the tumorigenesis of cells. The aim of this study was to determine the preclinical profile of a novel pan-Aurora kinase inhibitor, BPR1K653, as a candidate for anti-cancer therapy. Since expression of the drug efflux pump, MDR1, reduces the effectiveness of various chemotherapeutic compounds in human cancers, this study also aimed to determine whether the potency of BPR1K653 could be affected by the expression of MDR1 in cancer cells., Principal Findings: BPR1K653 specifically inhibited the activity of Aurora-A and Aurora-B kinase at low nano-molar concentrations in vitro. Anti-proliferative activity of BPR1K653 was evaluated in various human cancer cell lines. Results of the clonogenic assay showed that BPR1K653 was potent in targeting a variety of cancer cell lines regardless of the tissue origin, p53 status, or expression of MDR1. At the cellular level, BPR1K653 induced endo-replication and subsequent apoptosis in both MDR1-negative and MDR1-positive cancer cells. Importantly, it showed potent activity against the growth of xenograft tumors of the human cervical carcinoma KB and KB-derived MDR1-positive KB-VIN10 cells in nude mice. Finally, BPR1K653 also exhibited favorable pharmacokinetic properties in rats., Conclusions and Significance: BPR1K653 is a novel potent anti-cancer compound, and its potency is not affected by the expression of the multiple drug resistant protein, MDR1, in cancer cells. Therefore, BPR1K653 is a promising anti-cancer compound that has potential for the management of various malignancies, particularly for patients with MDR1-related drug resistance after prolonged chemotherapeutic treatments.

The rhizome of ginger (Zingiber officinale Roscoe) has been reputed to have many curative properties in traditional medicine, and recent publications have also shown that many agents in ginger possess anticancer properties. Here we show that the ethyl acetate fraction of ginger extract can inhibit the expression of the two prominent molecular targets of cancer, the human telomerase reverse transcriptase (hTERT) and c-Myc, in A549 lung cancer cells in a time- and concentration-dependent manner. The treated cells exhibited diminished telomerase activity because of reduced protein production rather than direct inhibition of telomerase. The reduction of hTERT expression coincided with the reduction of c-Myc expression, which is one of the hTERT transcription factors; thus, the reduction in hTERT expression might be due in part to the decrease of c-Myc. As both telomerase inhibition and Myc inhibition are cancer-specific targets for cancer therapy, ginger extract might prove to be beneficial as a complementary agent in cancer prevention and maintenance therapy.

Specifically tailored ω-alkenyl-1-carboxylic acids were synthesized for use as surfactants in the single-step preparation of manganese ferrite (MnFe2O4) nanoparticles (NPs). Monodisperse manganese ferrite NPs terminated with ω-alkenyl moieties were prepared via a one-pot reaction at high temperature without the need of ligand exchange. Using this approach, simple adjustment of the rate of heating allowed precise tuning of the size of the nanoparticles, which were characterized in bulk form by transmission electron microscopy (TEM), Fourier-transform infrared (FT-IR) spectroscopy, and X-ray diffraction (XRD). These surfactant-coated magnetic nanoparticles were then deposited onto hydrogen-terminated silicon(111) wafers and covalently anchored to the surface by UV-initiated covalent bonding. Analysis by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) confirmed that the UV treatment led to covalent immobilization of the NPs on the silicon surface with a consistent packing density across the surface. The magnetic properties of the stable, surface-bound nanoparticle arrays were characterized using a superconducting quantum interference device (SQUID) magnetometer. The materials and methods described here are being developed for use in bit-patterned ultrahigh density magnetic recording media and nanoscale biomagnetic sensing.

Background: Anti-mitotic compounds (microtubule de-stabilizers) such as vincristine and vinblastine have been shown clinically successful in treating various cancers. However, development of drug-resistance cells limits their efficacies in clinical situations. Therefore, experiments were performed to determine possible drug resistance mechanisms related to the application of anti-mitotic cancer therapy., Principal Findings: A KB-derived microtubule de-stabilizer-resistant KB-L30 cancer cell line was generated for this study. KB-L30 cells showed cross-resistance to various microtubule de-stabilizers including BPR0L075, vincristine and colchicine through multiple-drug resistant (MDR)-independent mechanisms. Surprisingly, KB-L30 cells showed hyper-sensitivity to the microtubule-stabilizer, paclitaxel. Results of the RT-PCR analysis revealed that expression of both class II and III β-tubulin was down-regulated in KB-L30 cells as compared to its parental KB cancer cells. In addition, DNA sequencing analysis revealed six novel mutation sites present in exon four of the βI-tubulin gene. Computational modeling indicated that a direct relationship exists between βI-tubulin mutations and alteration in the microtubule assembly and dynamic instability in KB-L30 cells and this predicted model was supported by an increased microtubule assembly and reduced microtubule dynamic instability in KB-L30 cells, as shown by Western blot analysis., Conclusions and Significance: Our study demonstrated that these novel mutations in exon four of the βI-tubulin induced resistance to microtubule de-stabilizers and hyper-sensitivity to microtubule stabilizer through an alteration in the microtubule assembly and dynamics in cancer cells. Importantly, the current study reveals that cancer cells may acquire drug resistance ability to anti-mitotic compounds through multiple changes in the microtubule networks. This study further provided molecular information in drug selection for patients with specific tubulin mutations.

Predicting the long-term potential of permeable reactive barriers for treating contaminated groundwater relies on understanding the endpoints of biogeochemical reactions between influent groundwater and the reactive medium. Iron hydroxy carbonate (chukanovite) is frequently observed as a secondary mineral precipitate in granular iron PRBs. Mineralogical characterization was carried out using X-ray diffraction, scanning electron microscopy, thermogravimetric analysis, and X-ray absorption spectroscopy on materials collected from three field-based PRBs in the US (East Helena, MT; Elizabeth City, NC; Denver Federal Center, CO). These PRBs were installed to treat a range of contaminants, including chlorinated organics, hexavalent chromium, and arsenic. Results obtained indicate that chukanovite is a prevalent secondary precipitate in the PRBs. Laboratory experiments on high-purity chukanovite separates were carried out to constrain the room-temperature solubility for this mineral. An estimated Gibbs energy of formation (Delta(f)G degrees) for chukanovite is -1174.4 +/- 6 kJ/mol. A mineral stability diagram is consistent with observations from the field. Water chemistry from the three reactive barriers falls inside the predicted stability field for chukanovite, at inorganic carbon concentrations intermediate to the stability fields of siderite and ferrous hydroxide. These new data will aid in developing better predictive models of mineral accumulation in zerovalent iron PRBs., (Published by Elsevier B.V.)

We report observations of phenomena within two-component Langmuir monolayers that are analogous to surfactant self-assembly in 3D solutions. A partially fluorinated fatty phosphonic acid played the role of 2D surfactant (linactant), and a perfluorinated fatty acid acted as the 2D solvent. Langmuir-Blodgett monolayers were prepared from mixtures of these compounds and examined using atomic force microscopy. Above a critical linactant mole fraction of approximately 0.013, distinctive monodisperse structural features were observed with a characteristic diameter of approximately 30 nm and a relative height of 1.4 nm. As the linactant concentration was further increased, the number density of these features increased linearly with concentration, whereas the size remained approximately the same. A quantitative analysis of these observations suggested that the features corresponded to self-limiting clusters composed of approximately 2000 linactant molecules and that the dispersed clusters represented a 2D micellar phase. Above a linactant mole fraction of 0.63, the clusters organized into a local hexagonal structure with short-range positional order indicative of a 2D "lyotropic" liquid crystalline phase; the correlation length increased systematically with increasing linactant concentration.

Protein phosphorylation plays a crucial role in the signal transduction pathways that regulate gene expression, metabolism, cell adhesion, and cell survival in response to oxidative stress. In this study, we have used hydrogen peroxide treatment of H9C2 rat cardiomyocytes as a model of oxidative stress in heart ischemia-reperfusion injury. We show that oxidative stress induces a robust tyrosine phosphorylation of multiple proteins in this cell type. A phosphoproteomics approach using anti-phosphotyrosine affinity purification and LC-MS/MS was then used to identify the protein targets of this stress-induced phosphorylation. Twenty-three tyrosine-phosphorylated proteins were identified, with the majority known to be associated with cell-cell junctions, the actin cytoskeleton, and cell adhesion. This suggested that oxidative stress may have a profound effect on intercellular connections and the cytoskeleton to affect cell adhesion, morphology, and survival. Importantly, Src kinase was shown to be a major upstream regulator of these events. Immunofluorescence studies, fluorescence-activated cell sorting, and cell-based assays were used to demonstrate oxidative stress-induced modification of cell adhesion structures and the cytoskeleton, induced de-adhesion, and increased apoptosis, which were reversed by treatment with the Src kinase inhibitor PP1. These data demonstrate the critical role of Src kinase in oxidative stress-induced phosphorylation and cell damage in cardiomyocytes and suggest that targeting this kinase may be an effective strategy for preventing ischemia-reperfusion injury in the heart., ((c) 2010 Elsevier Inc. All rights reserved.)

Background: It is essential to subculture the cells once cultured cells reach confluence. For this, trypsin is frequently applied to dissociate adhesive cells from the substratum. However, due to the proteolytic activity of trypsin, cell surface proteins are often cleaved, which leads to dysregulation of the cell functions., Methods: In this study, a triplicate 2D-DIGE strategy has been performed to monitor trypsin-induced proteome alterations. The differentially expressed spots were identified by MALDI-TOF MS and validated by immunoblotting., Results: 36 proteins are found to be differentially expressed in cells treated with trypsin, and proteins that are known to regulate cell metabolism, growth regulation, mitochondrial electron transportation and cell adhesion are down-regulated and proteins that regulate cell apoptosis are up-regulated after trypsin treatment. Further study shows that bcl-2 is down-regulated, p53 and p21 are both up-regulated after trypsinization., Conclusions: In summary, this is the first report that uses the proteomic approach to thoroughly study trypsin-induced cell physiological changes and provides researchers in carrying out their experimental design.

Although the extent of chemical-induced liver injury differs substantially from individual to individual, it is very hard to identify susceptible population priori to chemical exposure. We report here that the gene expression of the blood samples collected predose might identify the susceptible population without actual exposure to hepatotoxicant. The innate gene expressions in the blood samples collected at predose were compared using whole-genome microarray analysis and semiquantitative PCR with the extent of hepatotoxicity following the treatment of a model hepatotoxicant, carbon tetrachloride (CCl(4)) posteriori. The expression of 18 genes was found to innately differ in the blood of the susceptible animals from the resistant to CCl(4)-induced hepatotoxicity. Of these 18 genes, three genes, NADH dehydrogenase subunit 6 (ND6), transient receptor potential cation channel, subfamily C, member 6 (Trpc6), and tetraspanin 12 (Tspan12), were found to be different reproducibly in real-time PCR analysis with independent sets of animals. Of particular note, animals with the low expression level of ND6 and Tspan12 showed significantly higher susceptibility to CCl(4)-induced hepatotoxicity indeed. This study demonstrated that blood gene expression profiling might identify the susceptible individuals to chemical-induced hepatotoxicity without actual chemical exposure, providing a novel and important methodology for the prevention of drug-induced hepatotoxicity.

This paper describes the formation of a new series of monolayer films generated by the self-assembly of omega-cyclopropylalkanethiols, CyPr(CH(2))(n)SH (n = 9-13), onto the surface of gold. Procedures used to prepare the omega-cyclopropylalkanethiol adsorbates are also reported. Methyl-, vinyl-, and isopropyl-terminated self-assembled monolayers (SAMs) were also prepared and used as reference films to evaluate the structure and properties of the new cyclopropyl-terminated films. Ellipsometry and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) were used to examine the structure of the SAMs. A small but systematically lower thickness of the new films compared to that of analogous methyl-terminated SAMs was observed. Also, the orientation of the ring with respect to the surface normal was observed to vary systematically with the number of methylene groups in the adsorbate backbone (i.e., odd vs even chain lengths). Measurements of wettability by contact angle goniometry also revealed a small but reproducible "odd-even" effect for all contacting liquids used, except hexadecane, which almost completely wet the surfaces (theta(a) = 10-13 degrees ). When compared to the wettability data obtained from methyl- and isopropyl-terminated SAMs, the wettability data obtained from the cyclopropyl-terminated SAMs suggest that these films offer an increased density of atomic contacts per unit area across the surface, and thus enhanced attractive interactions with contacting liquids. Comparison of the wettabilities of vinyl-terminated and cyclopropyl-terminated films is complicated by dipole-induced dipole interactions and/or pi-pi interactions between the surfaces and the probe liquids. Furthermore, the significantly similar wettabilities of the cyclopropyl-terminated SAMs and the surface of polyethylene suggests that these SAMs (and perhaps other SAMs with judiciously designed tailgroups) can be used to mimic the interfacial properties of polymeric materials without complications arising from surface reconstruction.

The transformation of a normal cell into a cancer cell has been correlated with alterations in gene regulation and protein expression. To identify altered proteins and link them to the tumorigenesis of breast cancer, we have distinguished normal breast cells (MCF-10A) from noninvasive breast cancer cells (MCF-7) and invasive breast cancer cells (MB-MDA-231) to identify potential breast cancer markers in transformed breast cells. Using the 2D-DIGE and MALDI-TOF MS techniques, we quantified and identified differentially expressed extracellular secreted proteins and total cellular proteins across MCF-7, MB-MDA-231 and MCF-10A. The proteomic analysis of the secreted proteins identified 50 unique differentially expressed proteins from three different media. In addition, 133 unique differentially expressed proteins from total cellular proteins were also identified. Note that 14 of the secreted proteins and 51 of the total cellular proteins have not been previously reported in breast cancer research. Some of these unreported proteins have been examined in other breast cancer cell lines and have shown positive correlations with 2D-DIGE data. In summary, this study identifies numerous putative breast cancer markers from various stages of breast cancer. The results of this study may aid in developing proteins identified as useful diagnostic and therapeutic candidates in research on cancer and proteomics.

In the present work, we assessed the relationship between alkyl carbon chain length and ocular irritation potentials using the hen's egg test-chorioallantoic membrane (HET-CAM) and bovine corneal opacity and permeability (BCOP) assays using 5 commercial alkyl polyglucoside surfactants with different compositions of alkyl chain lengths (C(6)-C(16)). With HET-CAM, there was a good correlation between the proportion of C(10) alkyl polyglucoside and the eye irritation potential Q score (r(2) = 0.912, p = .011). There were no significant differences between the proportion of C(10) alkyl polyglucoside and corneal opacity in BCOP assays; however, there was a relatively high positive correlation between the proportion of C(10) alkyl carbon chain lengths and corneal permeability (r(2) = 0.736, p = .063).