Cariogenic potential of sweet flavors in electronic-cigarette liquids.

BACKGROUND: Most electronic-cigarette liquids contain propylene glycol, glycerin, nicotine and a wide variety of flavors of which many are sweet. Sweet flavors are classified as saccharides, esters, acids or aldehydes. This study investigates changes in cariogenic potential when tooth surfaces are exposed to e-cigarette aerosols generated from well-characterized reference e-liquids with sweet flavors. METHODS: Reference e-liquids were prepared by combining 20/80 propylene glycol/glycerin (by volume fraction), 10 mg/mL nicotine, and flavors. Aerosols were generated by a Universal Electronic-Cigarette Testing Device (49.2 W, 0.2 Ω). Streptococcus mutans (UA159) were exposed to aerosols on tooth enamel and the biological and physiochemical parameters were measured. RESULTS: E-cigarette aerosols produced four-fold increase in microbial adhesion to enamel. Exposure to flavored aerosols led to two-fold increase in biofilm formation and up to a 27% decrease in enamel hardness compared to unflavored controls. Esters (ethyl butyrate, hexyl acetate, and triacetin) in e-liquids were associated with consistent bacteria-initiated enamel demineralization, whereas sugar alcohol (ethyl maltol) inhibited S. mutans growth and adhesion. The viscosity of the e-liquid allowed S. mutans to adhere to pits and fissures. Aerosols contained five metals (mean ± standard deviation): calcium (0.409 ± 0.002) mg/L, copper (0.011 ± 0.001) mg/L, iron (0.0051 ± 0.0003) mg/L, magnesium (0.017 ± 0.002) mg/L, and silicon (0.166 ± 0.005) mg/L. CONCLUSIONS: This study systematically evaluated e-cigarette aerosols and found that the aerosols have similar physio-chemical properties as high-sucrose, gelatinous candies and acidic drinks. Our data suggest that the combination of the viscosity of e-liquids and some classes of chemicals in sweet flavors may increase the risk of cariogenic potential. Clinical investigation is warranted to confirm the data shown here.

Point-of-care production of therapeutic proteins of good-manufacturing-practice quality.

Manufacturing technologies for biologics rely on large, centralized, good-manufacturing-practice (GMP) production facilities and on a cumbersome product-distribution network. Here, we report the development of an automated and portable medicines-on-demand device that enables consistent, small-scale GMP manufacturing of therapeutic-grade biologics on a timescale of hours. The device couples the in vitro translation of target proteins from ribosomal DNA, using extracts from reconstituted lyophilized Chinese hamster ovary cells, with the continuous purification of the proteins. We used the device to reproducibly manufacture His-tagged granulocyte-colony stimulating factor, erythropoietin, glucose-binding protein and diphtheria toxoid DT5. Medicines-on-demand technology may enable the rapid manufacturing of biologics at the point of care.

Analysis of synthetic phenethylamine street drugs using direct sample analysis coupled to accurate mass time of flight mass spectrometry.

Serious health complications and fatal overdoses have brought phenethylamine, otherwise known as 2C, drug abuse to the public's attention. These compounds are 2C-X series analogs of mescaline. The name "2C" results from the two carbons in the ethyl chain. The substituents on these emerging drugs are constantly changed in order to avoid listing as controlled substances. These synthetic drugs are marketed as having affects similar to LSD and MDMA and are typically consumed sublingually via blotter paper. Twenty-six commercially available standards were analyzed using a direct sample analysis technique coupled to accurate mass time of flight mass spectrometry. Direct sample analysis is an ambient ionization technique employing direct charge transfer from nitrogen reagent gas resulting in ionization and in-source collision induced dissociation. Mass spectral analysis was used to determine the fragmentation patterns of the 2C compounds. While the mass spectral data for the compounds are similar in fragmentation, they also display differences that allow the analyst to distinguish which compound is present. The fragmentation patterns, high-resolution mass spectral data, and isotope ratios are used to qualitatively identify two blotter paper street samples that were extracted together. As these drugs are emerging substances of abuse, there are no generally accepted protocols for this analysis. Additionally, presumptive, or screening, tests have not been widely developed for these materials. Direct sample analysis allows for a quick screening of seized compounds resulting in a preliminary identification, which can then be analyzed and confirmed using other analytical techniques.

Universal electronic-cigarette test: physiochemical characterization of reference e-liquid.

BACKGROUND: Despite the rising health and safety concerns of e-cigarettes, a universal e-cigarette testing method is still in its early developmental stage. The aim of this study was to develop an e-liquid Reference Material that can be used to improve accuracy and reproducibility of research results, and advance health risk assessment of e-cigarette products. METHODS: E-liquid Reference Material was developed by purity assessment, gravimetric measurement, homogeneity testing, and stability testing with material and instrument traceability (adopted from ISO 35:2006E). RESULTS: Homogeneity tests showed e-liquid Reference Material requires ≥ 1 h rotation at a speed of 5 rpm to reach complete homogeneity. Stability tests showed homogeneity is intact for at least 2 weeks without secondary separation, and e-liquids are stable in 21 °C-50 °C thermocycling conditions up to 72 h. A change in the e-liquid color was first observed at day seven, and progressed to 2- and 16 - fold increase in absorbance by one and 6 months respectively. We found that e-liquids do not have inherent material instabilities such as immiscibility or secondary separation. However, discrepancies in concentration and composition arose mainly due to viscosity of propylene glycol and glycerin. Aerosol generated from the e-liquid Reference Material had 16 chemical-byproducts and was composed of ~634,000 particles of which 38% were Fine Particulate Matters (<0.5 µm in diameter). CONCLUSIONS: The efforts described here to create a standardized e-liquid Reference Material aim to provide unbiased and robust testing parameters that may be useful for researchers, the industry and government agencies. Additionally, the reference e-liquid could open a channel of conversation among different laboratories by providing the means of independent verification and validation while establishing a system of transparency and reproducibility in materials and methods.

Systems analysis in Cellvibrio japonicus resolves predicted redundancy of ß-glucosidases and determines essential physiological functions.

Degradation of polysaccharides forms an essential arc in the carbon cycle, provides a percentage of our daily caloric intake, and is a major driver in the renewable chemical industry. Microorganisms proficient at degrading insoluble polysaccharides possess large numbers of carbohydrate active enzymes (CAZymes), many of which have been categorized as functionally redundant. Here we present data that suggests that CAZymes that have overlapping enzymatic activities can have unique, non-overlapping biological functions in the cell. Our comprehensive study to understand cellodextrin utilization in the soil saprophyte Cellvibrio japonicus found that only one of four predicted ß-glucosidases is required in a physiological context. Gene deletion analysis indicated that only the cel3B gene product is essential for efficient cellodextrin utilization in C. japonicus and is constitutively expressed at high levels. Interestingly, expression of individual ß-glucosidases in Escherichia coli K-12 enabled this non-cellulolytic bacterium to be fully capable of using cellobiose as a sole carbon source. Furthermore, enzyme kinetic studies indicated that the Cel3A enzyme is significantly more active than the Cel3B enzyme on the oligosaccharides but not disaccharides. Our approach for parsing related CAZymes to determine actual physiological roles in the cell can be applied to other polysaccharide-degradation systems.

Visualization of Ambient Mass Spectrometry with the Use of Schlieren Photography.

This manuscript outlines how to visualize mass spectrometry ambient ionization sources using schlieren photography. In order to properly optimize the mass spectrometer, it is necessary to characterize and understand the physical principles of the source. Most commercial ambient ionization sources utilize jets of nitrogen, helium, or atmospheric air to facilitate the ionization of the analyte. As a consequence, schlieren photography can be used to visualize the gas streams by exploiting the differences in refractive index between the streams and ambient air for visualization in real time. The basic setup requires a camera, mirror, flashlight, and razor blade. When properly configured, a real time image of the source is observed by watching its reflection. This allows for insight into the mechanism of action in the source, and pathways to its optimization can be elucidated. Light is shed on an otherwise invisible situation.

Molecular Ionization-Desorption Analysis Source (MIDAS) for Mass Spectrometry: Thin-Layer Chromatography.

Molecular ionization-desorption analysis source (MIDAS), which is a desorption atmospheric pressure chemical ionization (DAPCI) type source, for mass spectrometry has been developed as a multi-functional platform for the direct sampling of surfaces. In this article, its utility for the analysis of thin-layer chromatography (TLC) plates is highlighted. Amino acids, which are difficult to visualize without staining reagents or charring, were detected and identified directly from a TLC plate. To demonstrate the full potential of MIDAS, all active ingredients from an analgesic tablet, separated on a TLC plate, were successfully detected using both positive and negative ion modes. The identity of each of the compounds was confirmed from their mass spectra and compared against standards. Post separation, the chemical signal (blue permanent marker) as reference marks placed at the origin and solvent front were used to calculate retention factor (Rf) values from the resulting ion chromatogram. The quantitative capabilities of the device were exhibited by scanning caffeine spots on a TLC plate of increasing sample amount. A linear curve based on peak are, R2 = 0.994, was generated for seven spots ranging from 50 to 1000 ng of caffeine per spot.

Characterization of a Direct Sample Analysis (DSA) Ambient Ionization Source.

Water cluster ion intensity and distribution is affected by source conditions in direct sample analysis (DSA) ionization. Parameters investigated in this paper include source nozzle diameter, gas flow rate, and source positions relative to the mass spectrometer inlet. Schlieren photography was used to image the gas flow profile exiting the nozzle. Smaller nozzle diameters and higher flow rates produced clusters of the type [H + (H(2)O)(n)](+) with greater n and higher intensity than larger nozzles and lower gas flow rates. At high gas flow rates, the gas flow profile widened compared with the original nozzle diameter. At lower flow rates, the amount of expansion was reduced, which suggests that lowering the flow rate may allow for improvements in sampling spatial resolution.