Risk assessment of unintentional phthalates contaminants in cosmetics.

A risk assessment was performed for three types of phthalates, benzyl butyl phthalate (BBP), dibutyl phthalate (DBP), and di(2-ethylhexyl)phthalate (DEHP) unintentionally contaminated in cosmetics. A total of 100 products of 8 types of cosmetics were analyzed employing gas chromatography-mass spectrometry (GC-MS). By applying the maximum detected values of phthalates based on the worst exposure cases, their systemic exposure dosage (SED) was calculated. Accordingly, DEHP was identified as the main unintentional phthalates contaminants (0.10-600.00 ppm) in the cosmetics, with an SED of 3.37 × 10-9-3.75 × 10-4 mg/kg/day. In the non-cancer risk assessment, a margin of safety (MOS ≥ 100, safe) of 1.28 × 104-1.42 × 109 was estimated. In the cancer risk assessment, the lifetime cancer risk (LCR ≤ 10-5, safe) was determined to be 8.81 × 10-12-9.79 × 10-7. Based on the results of both risk assessments, the levels of unintentional phthalates contaminants in cosmetics were deemed safe. Some phthalates are widely used as plasticizers and are essential for daily life; however, various toxicities, including endocrine disruption, have been reported. Therefore, even under these "worst case" assumptions, an adequate margin of safety is shown such that this might be a low priority for further work although exposure to unintentional phthalates contaminants through cosmetics should be considered as part of cumulative exposure.

Alternative skin sensitization prediction and risk assessment using proinflammatory biomarkers, interleukin-1 beta (IL-1ß) and inducible nitric oxide synthase (iNOS).

As an alternative to animal tests for skin sensitization potency and risk assessment, cell viability and biomarkers related to skin sensitization were analyzed in THP-1 human monocytic leukemia cells. Cell viabilities of 90% (CV90) and 75% (CV75) were determined for 24 selected test chemicals. Further biomarkers related to skin sensitization were also determined under equivalent comparative conditions. In cell viability analyses, potent skin sensitizers exhibited high cytotoxicity, but non-sensitizers did not display this tendency. In biomarker analyses, interleukin-I beta (IL-1ß), inducible nitric oxide synthase (iNOS), IL-1ß+iNOS, and THP-1 IL-1ß+Raw 264.7 IL-1ß were found to be suitable for prediction of skin sensitization potency following classification as either skin sensitizers or non-sensitizers (accuracies of 91.7%, 87.5%, 83.3%, and 82.6%, respectively). A significant positive correlation was found between biomarkers and skin sensitization potency, with a correlation coefficient (R) of 0.7 or more (correlation coefficients of 0.77, 0.72, 0.7, and 0.84, respectively). Finally, the skin sensitization potency effective threefold concentration (EC) 3% was predicted using a biomarker equation, with resulting prediction rates (match rate with actual data) of 58.3%, 54.2%, 62.5%, and 60.9%, respectively. The prediction accuracy for the EC3 value obtained from animal data was calculated as 83.3%, 79.2%, 79.2%, and 73.9%, respectively. Thus, these biomarkers, IL-1ß and iNOS, may be alternatively used to predict skin sensitization potency and risk assessment.

Quantitative structure-activity and quantitative structure-property relationship approaches as alternative skin sensitization risk assessment methods.

This study aimed to predict skin sensitization potency of selected chemicals by quantitatively analyzing their physicochemical properties by employing quantitative structure-activity relationship (QSAR) and quantitative structure-property relationship (QSPR) approaches as alternative risk assessment methods to animal testing. Correlations between effective concentration for a stimulation index of 3 (EC3) (%), the amount of a chemical required to elicit a threefold increase in lymph node cell proliferative activity (stimulation index, ≥3), were calculated using local lymph node assay (LLNA) and physicochemical properties of 212 skin sensitizers and 38 non-sensitizers were investigated. The correlation coefficients between melting point (MP) and EC3 and between surface tension (ST) and EC3 were 0.65 and 0.69, respectively. The correlation coefficient for MP + ST and EC3 was estimated to be 0.72. Thus, correlation coefficients between EC3 and MP, ST, and MP + ST reliably predicted the skin sensitization potential of the chemicals with sensitivities of 72% (126/175), 70% (122/174), and 73% (116/158); specificities of 77% (27/35), 69% (22/32), and 81% (26/32); and accuracies of 73% (153/210), 70% (144/206), and 75% (142/190), respectively. Our findings suggest that the EC3 value may be more accurately predicted using the ST values of chemicals as opposed to MP values. Thus, information on MP and ST parameters of chemicals might be useful for predicting the EC3 values as not only an alternative approach to animal testing, but as a risk assessment method for skin sensitization.

Risk assessment of lithium-ion battery explosion: chemical leakages.

Use of lithium-ion batteries has raised safety issues owing to chemical leakages, overcharging, external heating, or explosions. A risk assessment was conducted for hydrofluoric acid (HF) and lithium hydroxide (LiOH) which potential might leak from lithium-ion batteries. The inhalation no-observed-adverse-effect-level (NOAEL) for HF was 0.75 mg/kg/d. When a lithium-ion battery explodes in a limited space, HF emissions amount to 10-100 ppm. Assuming the worst-case scenario, the conversion rate was calculated to be 81.8 mg/m3, and the average daily dose (ADD) was 19.5 mg/kg/d. Consequently, the margin of exposure (MOE = NOAEL/ADD) was 0.034, a value which constitutes an unsafe inhalation exposure for HF. Conversely, skin toxicity NOAEL for LiOH was 41.35 mg/kg/d-. This LiOH value reflects the amount of lithium in the lithium-ion battery, which is generated upon contact between water and the electrolyte. The quantity of lithium in a mobile phone is approximately 295 mg, and systemic exposure dose (SED) was 4.92 mg/kg/d. Accordingly, the MOE (NOAEL/SED) value was 8.41, and skin exposure of LiOH was deemed as safe for humans. However, it is important that Energy Storage System batteries still require safety measures and technologies for next-generation batteries, to prevent any potential explosions of lithium-ion batteries.

IL-1α and IL-1ß as alternative biomarkers for risk assessment and the prediction of skin sensitization potency.

Potential biomarkers of skin sensitization in RAW264.7 mouse macrophages were investigated as alternatives to animal experiments and risk assessment. The concentrations that resulted in a cell viability of 90% (CV90) and 75% (CV75) were calculated by using a water-soluble tetrazolium salt (WST)-1 assay and used to analyze the skin sensitization potency of 23 experimental materials under equivalent treatment conditions. In addition, the expression of interleukin (IL)-1α, IL-1ß, IL-31, tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), prostaglandin E2 (PGE2), and cyclooxygenase-2 (COX-2) was analyzed utilizing Western blotting. In the cell viability analysis, skin sensitizers were generally more cytotoxic and exhibited increased skin sensitization potency. However, nonsensitizers did not show any marked cytotoxic tendency. Biomarker analysis demonstrated that IL-1α, IL-1ß, and the combination of IL-1α and IL-1ß (IL-1α + IL-1ß) predicted reliably skin sensitization potential (1) sensitivities of 94.4%, 83.3%, and 83.3%, specificities of 100%, 100%, and 100%, and (2) accuracies of 95.7%, 87%, and 87%, respectively. These observations correlated most reliably as indicators for skin sensitization potency. Data suggest that IL-1α and IL-1ß may serve as potential biomarkers for skin sensitization and provide an alternative method to animal experiments for prediction of skin sensitization potency and risk assessment.

Inhibition of di(2-ethylhexyl) phthalate (DEHP)-induced endocrine disruption by co-treatment of vitamins C and E and their mechanism of action.

The endocrine disrupting actions of di(2-ethylhexyl) phthalate (DEHP) on testicular functions are postulated to involve excess free radical generation. Thus the aim of this study was to examine the ability of antioxidant vitamins C and E to prevent DEHP-induced testicular disruption in male Sprague-Dawley (SD) rats. SD male rats were administered DEHP alone or DEHP with vitamin C and/or vitamin E for 30 days. DEHP alone increased the levels of testosterone (T) and reduced estradiol (E2) concentrations. Supplementation with antioxidant vitamins diminished or restored serum T levels noted in DEHP-treated rats to control values. In contrast vitamins C and E increased E2 levels to control in rats administered DEHP. Antioxidants significantly improved the decreased testicular levels of reduced glutathione and activity of superoxide dismutase compared to DEHP-treatment alone. Co-treatment of vitamins C and E also markedly improved the reduced epididymal sperm head counts and elevated levels of malondialdehyde (MDA) or 8-hydroxydeoxyguanosine (8-OHdG) induced by DEHP treatment. These results support the concept that the adverse actions of DEHP may be related to increased free radical generation while co-treatment with vitamins C and E significantly blocked the actions of DEHP on male testicular functions.

Risk assessment of N-nitrosodiethylamine (NDEA) and N-nitrosodiethanolamine (NDELA) in cosmetics.

N-nitrosamines and their precursors found in cosmetics may be carcinogenic in humans. Thus the aim of this study was to carry out risk assessment for N-nitrosamines (N-nitrosodiethanolamine [NDELA], N-nitrosodiethylamine [NDEA]) and amines (triethanolamine [TEA], diethanolamine [DEA]) levels in cosmetics determined using validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) procedures. NDELA and NDEA concentrations were present at levels of "not detected" (N.D.) to 596.5 µg/kg and N.D. to 40.9 µg/kg, respectively. TEA and DEA concentrations ranged from N.D. to 860 µg/kg and N.D. to 26.22 µg/kg, respectively. The nitrite concentration (3-2250 mg/l), number of nitrosating agents to a maximum 5, and pH (3.93-10.09) were also assessed. The impact of N-nitrosamine formation on the levels of TEA, DEA, nitrite, and other nitrosating agents was also examined. N-nitrosamine concentrations correlated with the number of nitrosating agents and nitrite concentrations. Data demonstrated that higher nitrite concentrations and a greater number of nitrosating agents increased NDELA and NDEA yields. Further, the presence of TEA and DEA exerted a significant influence on N-nitrosamine formation. Risk assessments, including the margin of exposure (MOE) and lifetime cancer risk (LCR) for N-nitrosamines and margin of safety (MOS) for amines, were calculated using product type, use pattern, and concentrations. Exposure to maximum amounts of NDELA and NDEA resulted in MOE > 10,000 (based upon the benchmark dose lower confidence limit 10%) and LCR <1 × 10-5, respectively. In addition, TEA and DEA concentrations in cosmetic samples resulted in MOS values >100. Therefore, no apparent safety concerns were associated with cosmetic products containing NDELA, NDEA, TEA, and DEA in this study. However, since amines and nitrosating agents produce carcinogenic nitrosamines, their use in cosmetics needs to be minimized to levels as low as technically feasible.

Non-cancer, cancer, and dermal sensitization risk assessment of heavy metals in cosmetics.

The heavy metal content of cosmetics may be a cause for concern in that exposure to these metals is associated with adverse consequences. Thus, the aim of this study was to assess consequences attributed to exposure to heavy metals in cosmetics as determined by non-cancer, cancer, and sensitization risks methodologies. The quantification and exposure assessments of aluminum (Al), chromium (Cr), manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), lead (Pb), mercury (Hg), cadmium (Cd), antimony (Sb), and titanium (Ti) were performed by inductively coupled plasma-mass spectrometry. The non-cancer risk assessment of Al, Cr3+, Mn, Fe, Co, Ni, Cu, Zn, Cd, Sb, and Ti in cosmetic samples resulted in a margin of safety (MOS) greater than 100 or a hazard index (HI) of less than 1. However, the probability of lifetime cancer risk (LCR) resulting from dermal exposure to heavy metals from cosmetics exceeded the acceptable risk levels (LCR > 10-5). An exposure-based sensitization quantitative risk assessment determined that the ratios of acceptable exposure level to consumers for Ni, Co, Cu, or Hg were above 1, suggesting an absence of skin-sensitizing potential. For an average daily user of lip cosmetics, the estimated intakes of heavy metals were within the acceptable daily intake (ADI). The percentage of heavy users for which metal intakes exceeded ADIs were 20.37% for Pb, 9.26% for Mn, 1.85% for Cr3+, and 1.85% for Cr6+, respectively. Data suggested that the heavy metals present in cosmetics do not appear to pose a serious risk to health. However, for heavy users of lip cosmetics, contamination with some heavy metals, such as Pb, Mn, and Cr needs to be minimized.

Formation and inhibition of N-nitrosodiethanolamine in cosmetics under pH, temperature, and fluorescent, ultraviolet, and visual light.

N-nitrosodiethanolamine (NDELA), a type of nitrosamine, is a possible human carcinogen that may form in cosmetic products. The aim of this study was to examine the formation and inhibition of NDELA through chemical reactions of secondary amines including mono-ethanolamine, di-ethanolamine (DEA), and tri-ethanolamine (TEA), and sodium nitrite (SN) under varying conditions such as pH, temperature, and fluorescent, ultraviolet (UV), and visual light (VIS) using liquid chromatography-mass spectroscopy. In a mixture of TEA and SN under acidic conditions pH 2, residual NDELA concentrations rose significantly under various storage conditions in the following order: 50°C > 40°C > UV (2 W/m2) > VIS (4000 lux) > fluorescent light > 25°C > 10°C. In a mixture of DEA and SN under the same acidic pH 2 conditions, NDELA formation was significantly elevated in the following order: UV (2 W/m2) > VIS (4000 lux) > 50°C > 40°C > fluorescent light > 25°C > 10°C. Inhibition of NDELA formation by d-mannitol, vitamin C (Vit C), or vitamin E (Vit E) was determined under varying conditions of pH, temperature, and fluorescent, UV, and VIS. At high concentrations of 100 or 1000 µg/ml, Vit E significantly decreased residual NDELA compared with control levels under acidic pH 2, but not under basic pH 6. Among various antioxidants, Vit E reacted more effectively with many nitrosating agents such as nitrate and nitrite found in cosmetic products. Therefore, to reduce NDELA, it is recommended that cosmetics be stored under cool/amber conditions and that Vit E or Vit C inhibitors of nitrosation be optimally added to cosmetic formulations at concentrations between 100 and 1000 µg/ml.

Risk assessment of zinc oxide, a cosmetic ingredient used as a UV filter of sunscreens.

Zinc oxide (ZnO), an inorganic compound that appears as a white powder, is used frequently as an ingredient in sunscreens. The aim of this review was to examine the toxicology and risk assessment of ZnO based upon available published data. Recent studies on acute, sub-acute, and chronic toxicities of ZnO indicated that this compound is virtually non-toxic in animal models. However, it was reported that ZnO nanoparticles (NP) (particle size, 40 nm) induced significant changes in anemia-related hematologic parameters and mild to moderate pancreatitis in male and female Sprague-Dawley rats at 536.8 mg/kg/day in a 13-week oral toxicity study. ZnO displayed no carcinogenic potential, and skin penetration is low. No-observed-adverse-effect level (NOAEL) ZnO was determined to be 268.4 mg/kg/day in a 13-week oral toxicity study, and a maximum systemic exposure dose (SED) of ZnO was estimated to be 0.6 mg/kg/day based on topical application of sunscreen containing ZnO. Subsequently, the lowest margin of safety (MOS) was estimated to be 448.2, which indicates that the use of ZnO in sunscreen is safe. A risk assessment was undertaken considering other routes of exposure (inhalation or oral) and major product types (cream, lotion, spray, and propellant). Human data revealed that MOS values (7.37 for skin exposure from cream and lotion type; 8.64 for skin exposure of spray type; 12.87 for inhalation exposure of propellant type; 3.32 for oral exposure of sunscreen) are all within the safe range (MOS > 1). Risk assessment of ZnO indicates that this compound may be used safely in cosmetic products within the current regulatory limits of 25% in Korea.

Time-resolved energy transduction in a quantum capacitor.

The capability to deposit charge and energy quantum-by-quantum into a specific atomic site could lead to many previously unidentified applications. Here we report on the quantum capacitor formed by a strongly localized field possessing such capability. We investigated the charging dynamics of such a capacitor by using the unique scanning tunneling microscopy that combines nanosecond temporal and subangstrom spatial resolutions, and by using Si(001) as the electrode as well as the detector for excitations produced by the charging transitions. We show that sudden switching of a localized field induces a transiently empty quantum dot at the surface and that the dot acts as a tunable excitation source with subangstrom site selectivity. The timescale in the deexcitation of the dot suggests the formation of long-lived, excited states. Our study illustrates that a quantum capacitor has serious implications not only for the bottom-up nanotechnology but also for future switching devices.

Risk Assessment and Risk Reduction of Ptaquiloside in Bracken Fern.

This study was conducted to determine the optimal boiling time to reduce ptaquiloside (PTA) and to carry out a risk assessment for PTA, a representative toxic substance found in bracken fern (BF; Pteridium aquilinum), which is frequently consumed as food in East Asian countries. High-performance liquid chromatography showed that the concentration of PTA in BF was reduced by up to 99% after boiling for 20 min. Risk assessment results showed that the cancer margin of exposure (MOE; ≥ 25,000 = safe) to PTA for an average daily exposure scenario after boiling BF for 20 min was considered safe. In addition, the non-cancer MOE (≥ 300 = safe) to PTA under an average daily exposure scenario after BF boiling for 20 min was considered safe. However, human exposure to PTA was considered unsafe under the non-boiled BF exposure and maximum daily exposure scenarios. Therefore, boiling BF for at least 20 min is recommended before consumption, to reduce exposure to PTA as much as possible.

Monitoring of vegetation impact due to trampling on Cadillac Mountain summit using high spatial resolution remote sensing data sets.

Cadillac Mountain--the highest peak along the eastern seaboard of the United States--is a major tourist destination in Acadia National Park, Maine. Managing vegetation impact due to trampling on the Cadillac Mountain summit is extremely challenging because of the large number of visitors and the general open nature of landscape in this fragile subalpine environmental setting. Since 2000, more intensive management strategies--based on placing physical barriers and educational messages for visitors--have been employed to protect threatened vegetation, decrease vegetation impact, and enhance vegetation recovery in the vicinity of the summit loop trail. The primary purpose of this study was to evaluate the effect of the management strategies employed. For this purpose, vegetation cover changes between 2001 and 2007 were detected using multispectral high spatial resolution remote sensing data sets. A normalized difference vegetation index was employed to identify the rates of increase and decrease in the vegetation areas. Three buffering distances (30, 60, and 90 m) from the edges of the trail were used to define multiple spatial extents of the site, and the same spatial extents were employed at a nearby control site that had no visitors. No significant differences were detected between the mean rates of vegetation increase and decrease at the experimental site compared with a nearby control site in the case of a small spatial scale (≤30 m) comparison (in all cases P > 0.05). However, in the medium (≤60 m) and large (≤90 m) spatial scales, the rates of increased vegetation were significantly greater and rates of decreased vegetation significantly lower at the experimental site compared with the control site (in all cases P < 0.001). Research implications are explored that relate to the spatial extent of the radial patterns of impact of trampling on vegetation at the site level. Management implications are explored in terms of the spatial strategies used to decrease the impact of trampling on vegetation.

Detecting vegetation cover change on the summit of Cadillac Mountain using multi-temporal remote sensing datasets: 1979, 2001, and 2007.

This study examines the efficacy of management strategies implemented in 2000 to reduce visitor-induced vegetation impact and enhance vegetation recovery at the summit loop trail on Cadillac Mountain at Acadia National Park, Maine. Using single-spectral high-resolution remote sensing datasets captured in 1979, 2001, and 2007, pre-classification change detection analysis techniques were applied to measure fractional vegetation cover changes between the time periods. This popular sub-alpine summit with low-lying vegetation and attractive granite outcroppings experiences dispersed visitor use away from the designated trail, so three pre-defined spatial scales (small, 0-30 m; medium, 0-60 m; and large, 0-90 m) were examined in the vicinity of the summit loop trail with visitor use (experimental site) and a site chosen nearby in a relatively pristine undisturbed area (control site) with similar spatial scales. Results reveal significant changes in terms of rates of vegetation impact between 1979 and 2001 extending out to 90 m from the summit loop trail with no management at the site. No significant differences were detected among three spatial zones (inner, 0-30 m; middle, 30-60 m; and outer, 60-90 m) at the experimental site, but all were significantly higher rates of impact compared to similar spatial scales at the control site (all p < 0.001). In contrast, significant changes in rates of recovery between 2001 and 2007 were observed in the medium and large spatial scales at the experimental site under management as compared to the control site (all p < 0.05). Also during this later period a higher rate of recovery was observed in the outer zone as compared to the inner zone at the experimental site (p < 0.05). The overall study results suggest a trend in the desired direction for the site and visitor management strategies designed to reduce vegetation impact and enhance vegetation recovery at the summit loop trail of Cadillac Mountain since 2000. However, the vegetation recovery has been rather minimal and did not reach the level of cover observed during the 1979 time period. In addition, the advantages and some limitations of using remote sensing technologies are discussed in detecting vegetation change in this setting and potential application to other recreation settings.

Risk Assessment of 5-Chloro-2-Methylisothiazol-3(2H)-One/2-Methylisothiazol-3(2H)-One (CMIT/MIT) Used as a Preservative in Cosmetics.

The mixture of 5-chloro-2-methylisothiazol-3(2H)-one (CMIT) and 2-methylisothiazol-3(2H)-one (MIT), CMIT/MIT, is a preservative in cosmetics. CMIT/MIT is a highly effective preservative; however, it is also a commonly known skin sensitizer. Therefore, in the present study, a risk assessment for safety management of CMIT/MIT was conducted on products containing 0.0015% of CMIT/MIT, which is the maximum MIT level allowed in current products. The no observed adverse effect level (NOAEL) for CMIT/MIT was 2.8 mg/kg bw/day obtained from a two-generation reproductive toxicity test, and the skin sensitization toxicity standard value for CMIT/MIT, or the no expected sensitization induction level (NESIL), was 1.25 µg/cm2/day in humans. According to a calculation of body exposure to cosmetics use, the systemic exposure dosage (SED) was calculated as 0.00423 mg/kg bw/day when leave-on and rinse-off products were considered. Additionally, the consumer exposure level (CEL) amounted to 0.77512 µg/cm2/day for all representative cosmetics and 0.00584 µg/cm2/day for rinse-off products only. As a result, the non-cancer margin of safety (MOS) was calculated as 633, and CMIT/MIT was determined to be safe when all representative cosmetics were evaluated. In addition, the skin sensitization acceptable exposure level (AEL)/CEL was calculated as 0.00538 for all representative cosmetics and 2.14225 for rinse-off products; thus, CMIT/MIT was considered a skin sensitizer when all representative cosmetics were evaluated. Current regulations indicate that CMIT/MIT can only be used at concentrations 0.0015% or less and is prohibited from use in other cosmetics products. According to the results of this risk assessment, the CMIT/MIT regulatory values currently used in cosmetics are evaluated as appropriate.

Risk Assessment of Triclosan, a Cosmetic Preservative.

Triclosan (TCS) is an antimicrobial compound used in consumer products. The purpose of current study was to examine toxicology and risk assessment of TCS based on available data. Acute toxicities of oral, transdermal and inhalation routes were low, and phototoxicity and neurotoxicity were not observed. Topical treatment of TCS to animal caused mild irritation. TCS did not induce reproductive and developmental toxicity in rodents. In addition, genotoxicity was not considered based on in vitro and in vivo tests of TCS. It is not classified as a carcinogen in international authorities such as International Agency for Research on Cancer (IARC). No-observed-adverse-effect level (NOAEL) was determined 12 mg/kg bw/day for TCS, based on haematoxicity and reduction of absolute and relative spleen weights in a 104-week oral toxicity study in rats. Percutaneous absorption rate was set as 14%, which was human skin absorption study reported by National Industrial Chemicals Notification and Assessment Scheme (NICNAS) (2009). The systemic exposure dosage (SED) of TCS has been derived by two scenarios depending on the cosmetics usage of Koreans. The first scenario is the combined use of representative cosmetics and oral care products. The second scenario is the combined use of rinse-off products of cleansing, deodorants, coloring products, and oral care products. SEDs have been calculated as 0.14337 mg/kg bw/day for the first scenario and 0.04733 mg/kg bw/day for the second scenario. As a result, margin of safety (MOS) for the first and second scenarios was estimated to 84 and 253.5, respectively. Based on these results, exposure of TCS contained in rinse-off products, deodorants, and coloring products would not pose a significant health risk when it is used up to 0.3%.

Identification of a peptide sequence that improves transport of macromolecules across the intestinal mucosal barrier targeting goblet cells.

In this study, we demonstrated that the CSKSSDYQC-peptide ligand which was identified from a random phage-peptide library through an in vivo phage display technique with rats could prominently improve the transport efficiency of macromolecules, such as large filamentous phage particles (M13 bacteriophage), across the intestinal mucosal barrier. Synthetic CSKSSDYQC-peptide ligands significantly inhibited the binding of phage P1 encoding CSKSSDYQC-peptide ligands to the intestinal mucosal tissue and immunohistochemical analysis showed that the CSKSSDYQC-peptide ligands could be transported across the intestinal mucosal barrier via goblet cells as their specific gateway. Thus, we inferred that CSKSSDYQC-peptide ligand might have a specific receptor on the goblet cells and transported from intestinal lumen to systemic circulation by transcytosis mechanism. These results suggest that CSKSSDYQC-ligand could be a promising tool for development of an efficient oral delivery system for macromolecular therapeutics in the carrier-drug conjugate strategy.

Detoxifying effect of pyridoxine on acetaminophen-induced hepatotoxicity via suppressing oxidative stress injury.

The detoxifying effect of pyridoxine against acetaminophen (APAP)-induced hepatotoxicity was investigated. HepG2 cells were co-treated with APAP and pyridoxine to compare with betaine or methionine for 24 h. LDH, ALT and AST activities were measured to determine direct cells damage in vitro and in vivo. Lipid peroxidation, antioxidant enzymes activity, and glutathione level were measured. Cytochrome c releaseand procaspase-3, cleaved caspase-3, Bcl-2, or Bax protein levels were measured to determine APAP-induced apoptotic cell death. Pyridoxine treatment significantly increased cell viability and decreased leakage of LDH activity against APAP-induced hepatotoxicity in HepG2 cells. ALT and AST activities were dose-dependently reduced by pyridoxine treatment compared to APAP-treated group. Significant increases in activities of GST and GPx were observed after co-treatment with APAP and pyridoxine. Although APAP-induced Nrf2 and HO-1 expression levels were gradually reduced in HepG2 cells by pyridoxine treatment, induction of antioxidant enzymes activities were dose-dependently increased. These protected effects of pyridoxine against APAP-induced hepatoxicity were closely associated with suppression of APAP-induced oxidative stress and apoptotic cell death in HepG2 cells. These data indicated that the protective action of pyridoxine against hepatic cell injuries was involved in the direct antioxidant activity which provides a pivotal mechanism for its potential hepatoprotective action.

Artificial neural network models for prediction of intestinal permeability of oligopeptides.

BACKGROUND: Oral delivery is a highly desirable property for candidate drugs under development. Computational modeling could provide a quick and inexpensive way to assess the intestinal permeability of a molecule. Although there have been several studies aimed at predicting the intestinal absorption of chemical compounds, there have been no attempts to predict intestinal permeability on the basis of peptide sequence information. To develop models for predicting the intestinal permeability of peptides, we adopted an artificial neural network as a machine-learning algorithm. The positive control data consisted of intestinal barrier-permeable peptides obtained by the peroral phage display technique, and the negative control data were prepared from random sequences. RESULTS: The capacity of our models to make appropriate predictions was validated by statistical indicators including sensitivity, specificity, enrichment curve, and the area under the receiver operating characteristic (ROC) curve (the ROC score). The training and test set statistics indicated that our models were of strikingly good quality and could discriminate between permeable and random sequences with a high level of confidence. CONCLUSION: We developed artificial neural network models to predict the intestinal permeabilities of oligopeptides on the basis of peptide sequence information. Both binary and VHSE (principal components score Vectors of Hydrophobic, Steric and Electronic properties) descriptors produced statistically significant training models; the models with simple neural network architectures showed slightly greater predictive power than those with complex ones. We anticipate that our models will be applicable to the selection of intestinal barrier-permeable peptides for generating peptide drugs or peptidomimetics.

Dietary hydroxycinnamates prevent oxidative damages to liver, spleen, and bone marrow cells in irradiation-exposed mice.

Dietary hydroxycinnamates are considered as attractive materials for radioprotection. This study explores whether hydroxycinnamates protect against γ-radiation-induced cellular damages and hematopoietic stem cell senescence. C57BL/6 mice were orally administered with each of caffeic acid, p-coumaric acid, and ferulic acid (20mg/kg body weight) once per three days for five times before exposure to total body radiation (5 Gy). Irradiation increased the activities of alanine amino transaminase and aspartate aminotransferase in blood serum but decreased the anti-oxidant defense enzyme activities in the liver and spleen tissues. Oral administration of the compounds almost completely prevented irradiation-mediated changes in these enzyme activities. The hydroxycinnamates also inhibited the irradiation-mediated increases in the mitochondrial superoxide anions of Lin-Sca-1+c-Kit+ (LSK) cells and CD150+CD48- LSK cells in the bone marrow. These results suggest that dietary hydroxycinnamates protect against irradiation-mediated oxidative damages of tissues and bone marrow progenitor cells.