Understanding the changes in the skin microbiome and their relationship to host skin factors during aging remains largely unknown. To better understand this phenomenon, we collected samples for metagenomic and host skin factor analyses from the forearm, buttock, and facial skin from 158 Caucasian females aged 20-24, 3034, 40-44, 50-54, 60-64, and 70-74 years. Metagenomics analysis was performed using 16S ribosomal RNA gene sequencing, whereas host sebocyte gland area, skin lipids, natural moisturizing factors, and antimicrobial peptides measurements were also performed. These analyses showed that skin bacterial diversity increased at all the skin sites with increasing age. Of the bacterial genera with an average relative abundance >1%, only Lactobacillus and Cutibacterium demonstrated a significant change (decrease) in abundance at all sampled skin sites with increasing age. Additional bacterial genera demonstrated significant age- and site-specific changes in abundance. Analysis of sebocyte area, natural moisturizing factors, lipids, and antimicrobial peptides showed an age-related decrease in sebocyte area and increases in natural moisturizing factors/antimicrobial peptides/ skin lipids, all of which correlated with changes in specific bacterial genera. In conclusion, the human skin microbiome undergoes age-associated alterations that may reflect underlying age-related changes in cutaneous biology.

The advent of electrochemical affinity assays and sensors evolved from pioneering efforts in the 1970s to broaden the field of analytes accessible to the selective and sensitive performance of electrochemical detection. The foundation of electrochemical affinity assays/sensors is the specific capture of an analyte by an affinity element and the subsequent transduction of this event into a measurable signal. This review briefly covers the early development of affinity assays and then focuses on advances in the past decade. During this time, progress on electroactive labels, including the use of nanoparticles, quantum dots, organic and organometallic redox compounds, and enzymes with amplification schemes, has led to significant improvements in sensitivity. The emergence of nanomaterials along with microfabrication and microfluidics technology enabled research pathways that couple the ease of use of electrochemical detection for the development of devices that are more user friendly, disposable, and employable, such as lab-on-a-chip, paper, and wearable sensors.

Background An important trend in the personal care industry involves the development of body wash products that not only clean the skin without damage but deposit conditioning ingredients to improve skin barrier function. Objective The objective of this study was to develop skin biomarker measures to quantify the treatment effects of body wash products. Methods We employed analysis of structural proteins (keratin 1,10,11 and involucrin), a natural moisturizing factor (pyrrolidone carboxylic acid) and an inflammatory mediator (IL-1ra/IL-1 alpha) from adhesive discs with dry skin grading, TEWL and capacitance measurements to compare the effects of direct application of petrolatum, a high petrolatum depositing body wash, and a regular body wash on dry leg skin in a standard leg-wash treatment protocol. Results High depositing body wash and petrolatum had positive effects on stratum corneum barrier function as judged by biomarker analysis, biophysical measurements and skin grading compared to the regular body wash product. Conclusions The results clearly indicate that a combination of biomarker and biophysical property measurements is effective for determining the skin benefits of moisturizing body wash products.

Pyrithione glucuronide (PTG) and 2-thiopyridine glucuronide (ThPG) have been reported to be the major urinary metabolites in multiple animal species following administration of zinc pyrithione (ZnPT). However, the formation of these metabolites has never been confirmed in humans. A simple and rugged ultra-high-performance liquid chromatography high resolution mass spectrometry (UHPLC-MS/HRMS) method was developed and validated for the quantification of PTG and ThPG to investigate human metabolism of pyrithione following topical application of ZnPT as a shampoo. A UHPLC-MS/HRMS method was required due to the matrix interferences that were observed with the typical industry standard HPLC/tandem mass spectrometry (LC-MS/MS) methodology based on nominal mass triple quadrupole (QQQ) platform approach. Using UPLC-MS/HRMS, both PTG and ThPG were identified in human urine following topical application of ZnPT. The presence of these human urinary metabolites of pyrithione are consistent with findings from earlier studies in multiple animal species and suggest the metabolism of pyrithione is similar amongst those mammalian species studied.

This is a story of the 100-year path for voltammetric electroanalytical chemistry from the serendipitous discovery of polarography by Jaroslav Heyrovsky in 1922 to the miniature biosensors of today. In spite of issues with the dropping mercury electrode (DME), polarography was dominant for almost 50 years due to the good quantitative results it produced. Then, significant developments led to today's methods with drastic improvements in detectable concentration and amount, selectivity, ease of use, and breadth of application. Important steps forward include the shift from the DME to solid electrodes, the strategic modification of electrode surfaces chemically and with membranes, electrochemistry in thin layers of solution with the associated decrease in sample amount from milliliters to microliters and below, liquid chromatography with electrochemical detection, more powerful and smaller instrumentation, microfabrication of electrodes, pulse techniques that improved concentration limits of detection by discriminating against double layer charging, spectroelectrochemistry for enhanced selectivity by electrochemically changing a spectroscopic signal, cyclic voltammetry for the general utility that makes it the work-horse of voltammetry, and biosensors that dramatically expanded the applicability of voltammetry through the use of nature's biological catalysts (enzymes) and capture agents (antibodies, aptamers).

1,2-Unsaturated pyrrolizidine alkaloids (PAs) are sometimes present in foods or herbal supplements/medicines as impurities and pose potential concerns for liver genotoxicity/carcinogenicity. PAs display a strong structure toxicity relationship, however, current regulatory approaches to risk assessment take the precautionary approach of assuming all PAs display the same potency as the most toxic congeners lasiocarpine (LAS) and riddelliine (RID). Here we explore the relative potencies of a series of structurally diverse PAs by measuring DNA adduct formation in vitro in a rat sandwich culture hepatocyte (SCH) cell system. The adducts generated are consistent with those identified in vivo as biomarkers of PA exposure and potential liver-tumor formation. DNA reactive PAs require metabolic activation to form intermediates that bind DNA, therefore, adduct formation is a direct reflection of reactive metabolite formation. Since the area under the concentration versus time curve (AUC) for the depletion of parent PA from the extracellular media is a measure of PA exposure, the ratio of adducts/AUC provides a measure of hepatocyte exposure to DNA-binding metabolites corresponding to an intrinsic potency for DNA adduct formation. Intrinsic potencies relative to potencies for LAS compare well with existing relative potency data further affirming that PA toxicity varies considerably with chemical structure.

The cationic surfactant diethyldialkylester dimethyl ammonium chloride (DEEDMAC) is an active ingredient in liquid fabric softeners and, as such, is disposed of down the drain after consumer use. A monitoring program was conducted across the continental United States to measure the concentration of DEEDMAC in the effluent and sludge from 41 wastewater treatment plants (WWTPs). The concentration in the effluent ranged from 32.4 to 2660ng/L, with a mean and standard deviation of 232±450ng/L. The concentration in the sludge ranged from 0.707 to 314mg/kg dw, with a mean and standard deviation of 29.2±50mg/kg dw. The distribution of measured effluent concentrations was combined with a distribution of mixing zone dilutions factors to predict the distribution of DEEDMAC concentrations in mixing zones and sediments under mean flow and 10-year, 7 consecutive day lowest flow (7Q10 low flow) conditions. Additionally, the distribution of measured sludge concentrations was combined with a distribution of land applied sludge volumes and US tilling practices to obtain a predicted distribution of DEEDMAC concentrations in sludge amended soils. The 90th percentile concentrations of DEEDMAC in mixing zones and sediments under mean flow conditions was 10.3ng/L and 451ng/kg, respectively. The 90th percentile concentration in sludge amended soils was 1.92mg/kg. These predicted exposure concentrations were compared to published eco-toxicity data and showed that DEEDMAC has a wide margin of safety and poses negligible ecologic risk to aquatic, sediment, or terrestrial compartments.

Amine oxide (AO) surfactants are used widely in North American household detergents resulting in > 44,000 mtons disposed down the drain annually. Due to AOs substantial down the drain disposal volume, wide dispersive use, and high aquatic toxicity, there is a need to evaluate ecological exposure and corresponding risk. This study refined the current knowledge regarding the fate of AO disposed down the drain through laboratory simulation studies to evaluate biodegradation in the sewer and during activated sludge wastewater treatment. A monitoring program which measured effluent AO concentrations for the dominant carbon chain lengths, C12 and C14, at 44 wastewater treatment plants (WWTP) across the continental US was also conducted. The study results were then used as input into probabilistic exposure models to predict US receiving stream concentrations. In three separate OECD 314A Sewer Water Die-Away studies AO was rapidly biodegraded with > 76% mineralized by study completion and the geometric mean of the primary biodegradation rates being 0.184 h−1. Two OECD 303A Activated Sludge WWTP Simulation studies showed rapid and complete biodegradation of AO with ≤ 0.09% of parent AO remaining in the effluent, ≤ 0.03% of parent AO sorbed to sludge solids, and > 97% complete mineralization of AO. Monitoring at US WWPTs confirmed low levels of AO in effluents with mean C12 and C14AO concentrations of 52.8 and 20.1 ng/L respectively. Based on the monitoring data, the 90th percentile concentrations of C12 and C14AO for 7Q10 low flow stream conditions were > 2 orders of magnitude lower than the predicted no effect concentrations indicating negligible aquatic risk from AO in US receiving streams. This study verifies that AO is safe for the aquatic environment even at the currently high usage volumes due to rapid biodegradation during transit through the sewer and wastewater treatment.

Alcohol sulfates (AS), alcohol ethoxysulfates (AES), linear alkyl benzenesulfonates (LAS) and methyl ester sulfonates (MES) are anionic surfactants that are widely used in household detergents and consumer products resulting in over 1 million tons being disposed of down the drain annually in the US. A monitoring campaign was conducted which collected grab effluent samples from 44 wastewater treatment plants (WWTPs) across the US to generate statistical distributions of effluent concentrations for anionic surfactants. The mean concentrations for AS, AES, LAS and MES were 5.03±4.5, 1.95±0.7, 15.3±19, and 0.35±0.13μg/L respectively. Since each of these surfactants consist of multiple homologues that differ in their toxicity, the concentration of each homologue measured in an effluent sample was converted into a toxic unit (TU) by normalizing to the predicted no effect concentration (PNEC) derived from high tier effects data (mesocosm studies). The statistical distributions of the combined TUs in the effluents were used in combination with distributions of dilution factors for WWTP mixing zones to conduct a US-wide probabilistic risk assessment for the aquatic environment for each of the surfactants. The 90th percentile level of TUs for AS, AES, LAS and MES in mixing zones were 1.89×10-2, 2.73×10-3, 2.72×10-2, and 3.65×10-5 under 7Q10 (lowest river flow occurring over a 7day period every 10years) low flow conditions. Because these surfactants have the same toxicological mode of action, the TUs were summed and the aquatic safety for anionic surfactants as a whole was assessed. At the 90th percentile level under the conservative 7Q10 low flow conditions the forecasted TUs were 4.21×10-2 which indicates that there is a significant margin of safety for the class of anionic surfactants in US aquatic environments.

OTNE [1-(1,2,3,4,5,6,7,8-octahydro-2,3,8,8-tetramethyl-2-naphthyl)ethan-1-one; trade name Iso E Super] is a fragrance ingredient commonly used in consumer products which are disposed down the drain. This research measured effluent and sludge concentrations of OTNE at 44 US wastewater treatment plants (WWTP). The mean effluent and sludge concentrations were 0.69 ± 0.65 μg/L and 20.6 ± 33.8 mg/kg dw respectively. Distribution of OTNE effluent concentrations and dilution factors were used to predict surface water and sediment concentrations and distributions of OTNE sludge concentrations and loading rates were used to predict terrestrial concentrations. The 90th percentile concentration of OTNE in US WWTP mixing zones was predicted to be 0.04 and 0.85 μg/L under mean and 7Q10 low flow (lowest river flow occurring over a 7 day period every 10 years) conditions respectively. The 90th percentile sediment concentrations under mean and 7Q10 low flow conditions were predicted to be 0.081 and 1.6 mg/kg dw respectively. Based on current US sludge application practices, the 90th percentile OTNE terrestrial concentration was 1.38 mg/kg dw. The probability of OTNE concentrations being below the predicted no effect concentration (PNEC) for the aquatic and sediment compartments was greater than 99%. For the terrestrial compartment, the probability of OTNE concentrations being lower than the PNEC was 97% for current US sludge application practices. Based on the results of this study, OTNE concentrations in US WWTP effluent and sludge do not pose an ecological risk to aquatic, sediment and terrestrial organisms.

The skin on the lower legs of 25 female subjects was evaluated first in the winter, and then again in the summer of the same subjects. Barrier function was determined by measuring transepidermal water loss (TEWL), and skin hydration and dryness were evaluated by electrical measurements (Corneometer ® CM825) and visual grading. Stratum corneum (SC) was sampled using 10 sequential D-Squame sampling discs and analyzed for 2-pyrrolidone-5-carboxylic acid (PCA), keratin-1,10,11, interleukin 1α (IL-1α), interleukin 1 receptor antagonist (IL-1ra), selected ceramides, cholesterol, cholesterol sulfate, and selected free fatty acids. TEWL as well as the visual dryness grades were significantly lower in the summer while hydration was higher. PCA was significantly higher in the summer as were the keratins. The ratio IL-1ra:IL-1α, an indicator of skin inflammation, was significantly lower in the summer. The amount of protein removed by the tape strips was also significantly lower in summer indicating better SC cohesion. Among the SC lipids measured, total ceramides, individual ceramides, total fatty acids, and cholesterol were higher in summer compared to winter. Stearic acid and cholesterol sulfate were not significantly different between winter and summer.

Amine oxide (AO) is a cationically charged surfactant at environmental pH and has previously been assessed in the OECD (Organization for Economic Cooperation and Development) High Production Volume (HPV) chemicals program. Typical of cationic chemicals, AO is highly aquatically toxic. In this study we vastly improve the knowledge of AO toxicity by developing acute Quantitative Structure Activity Relationships (QSARs) for an alga (Desmodesmus subspicatus), an invertebrate (Daphnia magna) and a fish (Danio rerio) using the appropriate array of OECD Test Guidelines. A chronic toxicity QSAR was also determined for the most sensitive taxon, Desmodesmus. Pure AO spanning the chain lengths of C8 to C16 were tested individually with trace analytical confirmation of exposures in all tests. The QSARs were all of high quality (R2 0.92-0.98) with slopes ranging from -0.338 to -0.484. QSARs were then used to normalize toxicity outcomes for a larger, previously published data set used in HPV, European REACH (Registration, Evaluation, and Authorization of Chemicals), and peer reviewed publications. Two additional species, Lemna gibba (macrophyte) and Ankistrodesmus falcatus (alga) were studied in exposures to dodecyl (C12) AO to provide sufficient taxonomic diversity to conduct a Species Sensitivity Distribution (SSD) analysis. The SSD 5th percentile hazardous concentration (HC5) to C12 AO was found to be 0.052mg/L which is similar to an existing AO 28-d, 3-community periphyton community bioassay normalized to C12 AO (No-observed-effect-concentration or NOEC=0.152mg/L). The statistical properties of the SSD was probed suggesting that new studies of additional taxa would be required that were at least 10-fold more sensitive than the most sensitive taxon to move the HC5 lower by a factor of 3. The overall AO hazard assessment suggests a large margin of safety relative to published environmental exposure data.

Systemic exposure was measured in humans after hair dyeing with oxidative hair dyes containing 2.0% (A) or 1.0% (B) [(14)C]-p-phenylenediamine (PPD). Hair was dyed, rinsed, dried, clipped and shaved; blood and urine samples were collected for 48 hours after application. [(14)C] was measured in all materials, rinsing water, hair, plasma, urine and skin strips. Plasma and urine were also analysed by HLPC/MS/MS for PPD and its metabolites (B). Total mean recovery of radioactivity was 94.30% (A) or 96.21% (B). Mean plasma Cmax values were 132.6 or 97.4 ng [(14)C]-PPDeq/mL, mean AUC(0-∞) values 1415 or 966 ng [(14)C]-PPDeq/mL*hr in studies A or B, respectively. Urinary excretion of [(14)C] mainly occurred within 24 hrs after hair colouring with a total excretion of 0.72 or 0.88% of applied radioactivity in studies A or B, respectively. Only N,N'-diacetylated-PPD was detected in plasma and the urine. A TK-based human safety assessment estimated margins of safety of 23.3- or 65-fold relative to respective plasma AUC or Cmax values in rats at the NOAEL of a toxicity study. Overall, hair dyes containing PPD are unlikely to pose a health risk since they are used intermittently and systemic exposure is limited to the detoxified metabolite N,N'-diacetyl-PPD.

The polycyclic musks, AHTN and HHCB are fragrance ingredients widely used in consumer products. A monitoring campaign was conducted and collected grab effluent and sludge samples at 40 wastewater treatment plants (WWTP) across the United States to understand their occurrence and statistical distribution in these matrices. AHTN concentration in effluent ranged from <0.05 μg/L (LOQ) to 0.44 μg/L with a mean and standard deviation of 0.18 ± 0.11 μg/L. HHCB concentrations in effluent ranged from 0.45 to 4.79 μg/L with a mean of 1.86 ± 1.01 μg/L. AHTN concentrations in sludge ranged from 0.65 to 15.0mg/kg dw (dry weight) with a mean and standard deviation being 3.69 ± 2.57 mg/kg dw, while HHCB sludge concentrations were between 4.1 and 91 mg/kg with a mean of 34.0 ± 23.1mg/kg dw. Measured concentrations of AHTN and HHCB were significantly correlated with each other in both effluent and sludge. The concentrations of HHCB in both effluent and sludge were approximately an order of magnitude higher than those for AHTN, consistent with 2011 usage levels. The highest measured effluent concentrations for both AHTN and HHCB were below their respective freshwater PNECs (predicted no effect concentrations), indicating a negligible risk to biological communities below WWTPs, even in the absence of upstream dilution. Moreover, the large number of effluents and sludges sampled provides a statistical distribution of loadings that can be used to develop more extensive probabilistic exposure assessments for WWTP mixing zones and sludge amended soils.

BACKGROUND:Barrier function is integral to the health of epithelial tissues. Currently, there is a broad need to develop and improve our knowledge with regard to barrier function for reversal of mild skin irritation and dryness. However, there are few in vitro models that incorporate modulations of both lipids and epidermal differentiation programs for pre-clinical testing to aid in the understanding of barrier health.OBJECTIVE:We have generated a reconstituted epidermis on a decellularized dermis (DED) and characterized its barrier properties relative to human epidermis in order to determine its utility for modeling barrier formation and repair.METHODS:We followed the process of epidermal differentiation and barrier formation through immunocytochemistry and transcriptional profiling. We examined barrier functionality through measurements of surface pH, lipid composition, stratum corneum water content, and the ability to demonstrate topical dose-dependent exclusion of surfactant.RESULTS:Transcriptional profiling of the epidermal model during its formation reveals temporal patterns of gene expression associated with processes regulating barrier function. The profiling is supported by gradual formation and maturation of a stratum corneum and expression of appropriate markers of epidermis development. The model displays a functional barrier and a water gradient between the stratum corneum and viable layers, as determined by confocal Raman spectroscopy. The stratum corneum layer displays a normal acidic pH and an appropriate composition of barrier lipids.CONCLUSION:The epidermal model demonstrates its utility as an investigative tool for barrier health and provides a window into the transcriptional regulation of multiple aspects of barrier formation.