Aerosolized e-liquid base constituents induce cytotoxicity and genotoxicity in oral keratinocytes.

OBJECTIVE: Electronic cigarette (e-cigarette) use among adults in the United States continues to rise. Particularly concerning is the impact of e-cigarette aerosol inhalation on the oral mucosa. Aerosols are derived from a heated e-liquid base of propylene glycol/glycerin (PG/G) often mixed with nicotine and chemical flavors. Of note, harmful and potentially harmful constituents (HPHCs), including metals and volatile organic compounds, have been detected in e-cigarette aerosols. It remains unknown, however, whether aerosols exclusively derived from e-liquid PG/G are detrimental to oral keratinocytes. The present study analyzed toxicological outcomes in normal oral keratinocytes exposed to model nicotine-free, unflavored PG/G e-liquid aerosols. MATERIALS AND METHODS: Cell viability/cytotoxicity, genotoxicity, and immunoblotting assays were conducted in NOKSI, a gingiva-derived oral keratinocyte cell line, following exposure to model e-liquid aerosols or non-aerosolized controls. The HPHC acrolein, reported to form DNA adducts in the buccal mucosa from e-cigarette users, was also used in similar assays. RESULTS: PG/G e-liquid aerosol extracts significantly enhanced cytotoxic and DNA damaging responses in NOKSI cells when compared to non-aerosolized e-liquid treatment. Acrolein treatment led to similar results. CONCLUSIONS: The aerosolization process of PG/G e-liquid is a critical determinant of marked cytotoxic and genotoxic stimuli in oral keratinocytes.

Packaging of disposable vaping products and e-liquids in England, Canada and the United States: A content analysis.

BACKGROUND AND AIMS: Vaping product packaging is varied and often features bright colours and novel designs, particularly among recently marketed disposable vapes. This study provides an overview of attributes found on the packaging of popular disposable vapes and e-liquid bottles in England, Canada and the United States (US) and assesses compliance with local packaging regulations. DESIGN: Content analysis. SETTING: Brick-and-mortar and online shops in England (London), Canada (Ontario) and the US (New Hampshire and South Carolina). CASES: 108 vaping products (including packaging) from 76 brands in a range of flavours and nicotine levels. Specifically, 48 disposable vapes (15 from England, 16 from Canada, 17 from the US) and 60 e-liquid bottles (20 per country). MEASUREMENTS: Textual and graphic branding and marketing elements, independently coded by two researchers and checked by a third. FINDINGS: Compliance with local packaging regulations varied across countries. Health warnings were present on the packaging of all but one nicotine-containing product, although 33% of disposables and 17% of e-liquids featuring the warning did not adhere to formatting requirements. Leaflets were seldom included with e-liquid bottles, even in England (45%) where mandatory, and omitted elsewhere. Labelling of nicotine type and batch numbers was inconsistent. Vaping product packaging featured claims relating to sensory perceptions (41%), most often flavours, and some (32%) featured youth-appealing content. Common graphic elements included stylised brand fonts (80%), brand logos (54%), product representations on the external packaging (47%) and abstract graphic elements (64%). Colours on packaging, disposable vapes and e-liquid bottles were associated with product flavour. CONCLUSIONS: In England, Canada and the United States, popular disposable vapes and e-liquid bottles appear to have varying compliance with local packaging regulations and inconsistent labelling of nicotine and product characteristics. The use of colourful designs, evocative descriptors and appealing graphics to promote flavours underscores the need for comprehensive packaging regulations and enforcement.

Exploring the opinions and potential impact of unflavoured e-liquid on smoking cessation among people who smoke and smoking relapse among people who previously smoked and now use e-cigarettes: findings from a UK-based mixed methods study.

BACKGROUND: Although electronic cigarettes (e-cigarettes) appear to be effective in helping people who smoke to stop smoking, concerns about use of e-cigarettes among young people have led to restrictions on non-tobacco flavoured e-liquids in some countries and some US states. These restrictions could reduce the appeal of these products to non-smoking youth but could have negative consequences for people who smoke or use e-cigarettes. METHODS: In this mixed methods study, we recruited UK adults who smoked or used to smoke and subsequently vaped to explore their opinions of unflavoured e-liquids and their beliefs about how they would be impacted by hypothetical e-liquid flavour restrictions. Participants trialled an unflavoured e-liquid instead of their usual nicotine product for four hours and completed a survey and an online interview. RESULTS: Using Interpretive Phenomenological Analysis and graphically presented data, we found differences in participants' opinions of unflavoured e-liquid. If only unflavoured, tobacco flavoured, and menthol flavoured e-liquids remained on the UK market, some people who smoke or vape may be unaffected, but some may relapse to smoking or continue smoking. Despite most wanting to prevent young people from initiating vaping, participants had varying opinions on whether flavour restrictions would be an effective method. CONCLUSIONS: The findings highlight that people who smoke and vape could be impacted by flavour restrictions in a range of ways, some of which could have a potential adverse impact on harm reduction efforts in the UK (e.g., by making smoking more appealing than vaping).

Vaping: Public Health, Social Media, and Toxicity.

This viewpoint aims to provide a comprehensive understanding of vaping from various perspectives that contribute to the invention, development, spread, and consequences of e-cigarette products and vaping. Our analysis showed that the specific characteristics of e-cigarette products as well as marketing strategies, especially social media marketing, fostered the spread of vaping and the subsequent effects on human health and toxicity. We analyzed the components of e-cigarette devices and e-liquids, including the latest variants whose impacts were often overlooked. The different forms of nicotine, including salts and freebase nicotine, tobacco-derived nicotine, tobacco-free nicotine, and cooling agents (WS3 and WS23), have brought more choices for vapers along with more ways for e-cigarette manufacturers to advertise false understandings and present a greater threat to vapers' health. Our work emphasized the products of brands that have gained significant influence recently, which are contributing to severe public health issues. On the other hand, we also discussed in detail the toxicity of e-liquid components and proposed a toxicity mechanism. We also noticed that nicotine and other chemicals in e-liquids promote each other's negative effects through the oxidative stress and inflammatory nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway, a mechanism leading to pulmonary symptoms and addiction. The impact of government regulations on the products themselves, including flavor bans or regulations, has been limited. Therefore, we proposed further interventions or harm reduction strategies from a public health perspective.

Ecotoxicological effects of leachate from e-cigarettes and e-liquid on the performance of perennial ryegrass (Loliumperenne).

Once littered, disposable e-cigarettes present a complex type of waste in the environment. They typically contain a lithium battery, electronics to produce vapour and remnant e-liquid, all of which could leach into the environment. The effects of littered e-cigarettes are not well understood, and they have not been tested in terrestrial ecosystems. To address this, an experiment was set up to assess how leachate from e-cigarettes with or without a battery, but also e-liquid on its own can alter fundamental physical characteristics of Lolium perenne (perennial ryegrass) when irrigated with contaminated water. After 31 days, shoot length of L. perenne was not measurably affected, but the biomass was significantly reduced by 30% when e-liquid, and 24% when leachate from intact e-cigarettes was present compared to control plants. Plants grown with leachate or e-liquid displayed a significant level of early senescence of leaf apices, and the chlorophyll content was increased. Furthermore, root biomass was significantly less (29-46%) compared to the control. Leachate from used disposable e-cigarettes can affect the performance of plants when entering the soil ecosystem, therefore stricter regulations are needed to prevent this new type of electronic litter from becoming more widespread.

Recent advances in the analysis of electronic cigarette liquids and aerosols: Sample preparation and chromatographic characterization.

Electronic cigarette (e-cigarette) usage has risen dramatically worldwide in recent years. It has been publicized as a safer alternative to the conventional combustible cigarette. This, however, has not yet been supported by robust toxicological research evidence. Analysis of the chemical compositions of e-liquids and generated aerosols is an important step in evaluating the toxicity effects of e-cigarettes. Currently, a broad spectrum of analytical methods have been employed for qualitative and quantitative analysis of chemical compositions of e-cigarette liquids and aerosols. The aim of this article is to review the advances in the chromatographic characterization of chemical composition of the latter in the recent five years. In addition, sample preparation methods for e-liquids and aerosols are surveyed and discussed. A study of the relevant literature indicates that, expectedly, gas chromatography and liquid chromatography with a variety of detection systems, particularly mass spectrometry, have been the main analytical techniques used in this field. Sample preparation procedures primarily include headspace sampling, dilute-and-shoot approach, liquid-liquid extraction and sorbent-based extraction for e-liquids and for aerosols (the latter usually with laboratory-built collection devices). Some challenges of current e-cigarette analytical research, and an overview on prospective work are also presented.

Photometric Monitoring of Electronic Cigarette Puff Topography.

To study and monitor the adverse health consequences of using electronic cigarettes, a user's puff topography, which are quantification parameters of the user's vaping habits, plays a central role. In this work, we introduce a topography sensor to measure the mass of total particulate matter generated in every puff and to estimate the nicotine yield. The sensor is compact and low-cost, and is integrated into the electronic cigarette device to promptly and conveniently monitor the user's daily puff topography. The topography sensor is comprised of a photometric sensor and a pressure sensor. The photometric sensor measures the mass concentration of the aerosol, based on scattering of near-infrared light from airborne particles, while the pressure sensor measures the flow rate. The topography sensor was tested under various conditions including a wide range of atomizer power, puff duration, and inhalation pressure. The sensor's accuracy was validated by comparing the sensor's readings with reference measurements, and the results matched closely with the trends reported by existing studies on electronic cigarettes. An example application for tracking a user's puff topography was also demonstrated. Our topography sensor holds great promise in mitigating the health risks of vaping, and in promoting quality control of electronic cigarette products.

Hypothetical e-liquid flavor ban and opinions among vape shop retailers in the Greater Los Angeles Area.

INTRODUCTION: Evaluating anticipated responses to flavor bans in the context of vape shops is needed to inform legislation and enforcement. This cross-sectional study examined vape shop retailers' opinions about the potential impacts of an e-liquid flavor ban on shop sales and customer behavior-change intentions. METHODS: From December 2019 to October 2020 we conducted structured interviews over the phone with 46 brick-and-mortar vape shop retailers in the Greater Los Angeles Area. RESULTS: Most participants were managers (43.5%), followed by owners (26.1%) and clerks (26.1%). More than half (52.2%) reported that sales would drop a lot if flavored e-liquids were banned in all vape shops. Controlling for store position, multivariable linear regression showed that opposition to a hypothetical ban on non-tobacco flavored e-liquids was associated with participants' opinions that customers would likely not purchase tobacco flavored e-liquids (b= -0.44, p<0.01), and would likely use combustible tobacco products (b=0.47, p<0.05). CONCLUSIONS: In this cross-sectional study, vape shop retailers in the Greater Los Angeles Area reported that if a ban on non-tobacco e-liquid flavors occurred, they would oppose strongly, and that a ban would have a negative impact on their shop (e.g. loss in sales) and customer behavior (e.g. would replace vaping with smoking combustible tobacco products). Implications for research and practice are discussed.

Impact of e-cigarette liquid on porcine lung tissue-Ex vivo confocal Raman micro-spectroscopy study.

Ex vivo porcine lung immersed in e-liquid was investigated in-depth using confocal Raman micro-spectroscopy to assess the e-liquid influence on the lung. It was found that lung-related Raman band intensities at 1002, 1548, 1618 and 1655 cm-1 increased after first and second treatments except the surface, which was attributed to the well-known optical clearing (OC) effect due to alveoli filling with e-liquid resulting in light scattering reduction. The autofluorescence enhancement was explained by oxidative stress induced in lung during exposure to e-liquid. Moreover, e-liquid induced collagen dehydration was revealed by the I937 /I926 Raman band intensity ratio change. The effect was enhanced after the second treatment of the same lung tissue that indicates the possibility of multi-step OC treatment. We hypothesize that the nicotine-flavour-free e-liquids containing glycerol and propylene glycol could potentially be used in clinical protocols as OC agent for enhanced in-depth Raman-guided bronchoscopy.

In Vitro high-throughput toxicological assessment of E-cigarette flavors on human bronchial epithelial cells and the potential involvement of TRPA1 in cinnamon flavor-induced toxicity.

Electronic cigarettes (ECs) are considered a less hazardous alternative to tobacco smoking but are not harmless. Growing concerns about the safety profiles of flavors in e-liquids underpin the need for this study. Here, we screened 53 nicotine-free flavored e-liquids (across 15 flavor categories) across a 3-point concentration range (0.25%, 0.5%, and 1% v/v) in a high-throughput fashion in human bronchial epithelial (HBEC-3KT) submerged cell cultures to identify 'toxic hits' using in vitro endpoint assays comprising cell count, cell viability, and lactate dehydrogenase (LDH). We observed significant, dose-dependent adverse effects only with cinnamon, vanilla tobacco, and hazelnut e-liquids compared to media-only and PG/VG vehicle controls. Hence, we further analyzed these three flavors for their effects on HBEC-3KT proliferation, mitochondrial health, and oxidative stress. A significant decrease in cell proliferation after 36 h was observed for each e-liquid toxic hit compared to media-only and PG/VG controls. Hazelnut (at all concentrations) and vanilla tobacco (1%) increased cytoplasmic reactive oxygen species generation compared to media-only and PG/VG controls. Conversely, all three flavors at 0.5% and 1% significantly decreased mitochondrial membrane potential compared to PG/VG and media-only controls. Chemical analysis revealed that all three flavors contained volatile organic compounds. We hypothesized that the cytotoxicity of cinnamon might be mediated via TRPA1; however, TRPA1 antagonist AP-18 (10 µM) did not mitigate these effects, and cinnamon significantly increased TRPA1 transcript levels. Therefore, pathways mediating cinnamon's cytotoxicity warrant further investigations. This study could inform public health authorities on the relative health risks assessment following exposure to EC flavor ingredients.

Nicotine and Microvascular Responses in Skeletal Muscle from Acute Exposure to Cigarettes and Vaping.

Despite claims of safety or harm reduction for electronic cigarettes (E-cig) use (also known as vaping), emerging evidence indicates that E-cigs are not likely safe, or necessarily safer than traditional cigarettes, when considering the user's risk of developing vascular dysfunction/disease. E-cigs are different from regular cigarettes in that E-cig devices are highly customizable, and users can change the e-liquid composition (such as the base solution, flavors, and nicotine level). Since the effects of E-cigs on the microvascular responses in skeletal muscle are poorly understood, we used intravital microscopy with an acute (one-time 10 puff) exposure paradigm to evaluate the individual components of e-liquid on vascular tone and endothelial function in the arterioles of the gluteus maximus muscle of anesthetized C57Bl/6 mice. Consistent with the molecular responses seen with endothelial cells, we found that the peripheral vasoconstriction response was similar between mice exposed to E-cig aerosol or cigarette smoke (i.e., 3R4F reference cigarette); this response was not nicotine dependent, and endothelial cell-mediated vasodilation was not altered within this acute exposure paradigm. We also report that, regardless of the base solution component [i.e., vegetable glycerin (VG)-only or propylene glycol (PG)-only], the vasoconstriction responses were the same in mice with inhalation exposure to 3R4F cigarette smoke or E-cig aerosol. Key findings from this work reveal that some component other than nicotine, in inhaled smoke or aerosol, is responsible for triggering peripheral vasoconstriction in skeletal muscle, and that regardless of one's preference for an E-cig base solution composition (i.e., ratio of VG-to-PG), the acute physiological response to blood vessels appears to be the same. The data suggest that vaping is not likely to be 'safer' than smoking towards blood vessels and can be expected to produce and/or result in the same adverse vascular health outcomes associated with smoking cigarettes.

Effect of Sample Plates and Sample Matrix on the Quantification Capabilities of Surface-Assisted Flowing Atmospheric-Pressure Afterglow Mass Spectrometry (SA-FAPA-MS).

Ambient desorption/ionization mass spectrometry (ADI-MS) has been broadly applied to accomplish direct analysis without sample preparation or separation. However, quantification capabilities and analytical performance are sometimes limited. Here, we report signal enhancement effects and improved quantification capabilities in plasma-based ADI-MS, when a flowing atmospheric-pressure afterglow (FAPA) source is used to probe analytes on tailored thin-layer chromatography (TLC) plates. It was found that quantitative results could be achieved when the TLC plate merely served as a sampling plate without a preceding separation step. Specifically, the dynamic response of caffeine, nicotine, acetaminophen, and progesterone was investigated with FAPA-MS on a variety of different TLC surfaces (normal-phase silica, reversed-phase-modified silica, cyano [CN]-modified silica, and dimethyl [RP2]-modified silica). All analytes were studied as single-analyte standards and in a multianalyte mixture to evaluate the effect of sample plates and sample matrix on analytical performance and competitive ionization processes. Overall, dimethyl (RP2)- and CN-modified silica resulted in superior performance compared to other TLC materials. After careful optimization and without the use of internal standards, linear ranges of five orders of magnitude were accessible for caffeine and nicotine. Limits of detection down to femtomole amounts of analyte were achieved. Quantitation limits using RP2-TLC and FAPA-MS were 0.062, 0.062l, 0.31, and 14 pmol for caffeine, nicotine, progesterone, and acetaminophen, respectively. Interestingly, the presence of nicotine at relatively high amounts reduced the signal of the other analytes, an observation that was found to correlate with the differences in the enthalpy of vaporization (ΔHvap) and proton affinity. To prove the quantitative capabilities, nicotine quantification in a real matrix-heavy e-liquid sample was demonstrated using an isotopically labeled standard. The use of TLC-based surfaces with FAPA-MS can aid in the direct and quantitative mass spectrometric investigation of complex mixtures.

Synthetic nicotine e-liquids sold in US online vape shops.

Synthetic nicotine (relative to tobacco-derived, or "natural" nicotine) is an emerging feature of e-cigarettes including e-liquids in the online marketplace. This study investigated a total of 11,161 unique nicotine e-liquids sold in online stores in the US during 2021, using keyword matching approach to identify the feature of synthetic nicotine based on product description texts. We showed that in 2021, 2.13% of nicotine-containing e-liquids in our sample were marketed as synthetic nicotine e-liquids. About a quarter of the synthetic nicotine e-liquids that we identified were salt-based; the nicotine strength varied; and those synthetic nicotine e-liquids had a variety of flavor profiles. Synthetic nicotine containing e-cigarettes are likely to remain in the market and manufacturers might market those products as "tobacco-free," to attract consumers who this feature as healthier or less addictive. It is important to monitor synthetic nicotine in the e-cigarette marketplace and assess how this feature influences consumer behaviors.

Effects of Device Settings and E-Liquid Characteristics on Mouth-Throat Losses of Nicotine Delivered with Electronic Nicotine Delivery Systems (ENDS).

Currently it is not fully understood how the device settings and electronic liquid (e-liquid) composition, including their form of nicotine content, impact mouth and throat losses, and potentially lead to the variations in total nicotine delivery to the human lungs. An in situ size assessment method was developed for real-time measurements at the mouthpiece and outlet of a biorelevant mouth-throat to account for the dynamic nature of the aerosol. The aerosol size, temperature, and delivery through the mouth-throat replica and the exhaled aerosol between the puff intervals were measured at different wattages using various e-liquid compositions. The effects of body temperature and humidity on aerosol size and nicotine delivery were also explored to evaluate the importance of considering realistic in vivo conditions in in vitro measurements. Notably, in vitro tests with body temperature and humidity in mouth-throat model vs room conditions, resulted in larger aerosol size at the end of the throat (Dv50=5.83±0.33 µm vs 3.05±0.15 µm), significantly higher thoracic nicotine delivery (>90% vs 50-85%) potentially due to the lower exhaled amount (<10% vs 15-50%). Besides, higher VG/PG ratios resulted in significantly lower exhaled amount and higher mouth-throat nicotine deposition. One of the main outcomes of the study was finding significantly lower exhaled amount and higher thoracic nicotine delivery with nicotine salt form vs free-base. Considering body temperature and humidity also showed significant enhancement in nicotine delivery, so it is essential to account for biorelevant experimental conditions in benchtop testing.

Profiling flavourings in strawberry-flavoured e-liquid using headspace-gas chromatography-mass spectrometry.

E-liquids typically contain nicotine and flavourings in a matrix of propylene glycol (PG) and vegetable glycerine (VG). Some nicotine-free e-liquids are flavouring only in the aerosol carrier with the option for users to add their own nicotine. It is only the nicotine that is monitored in terms of level, as specified by the manufacturers. Little is known of the toxicological effect for some of the flavourings in the context of vaping as these are only regulated for ingestion and not inhalation. A method was developed to analyse volatile organic compounds (VOCs) evolved when e-liquids are vaporised based on headspace-gas chromatography-mass spectrometry (HS-GC-MS) for e-liquids. An in-house standard was prepared with sample matrix and purchased strawberry flavouring to simulate a simple e-liquid but with known levels. This standard was then used to optimise the analysis for use with e-liquid samples but not for full quantification purposes. These were purchased from a range of retailers and with different batches but mainly focussed on strawberry flavour. The results identified three key components indicative of strawberry flavour (ethyl-3-methyl butanoate, ethyl 2-methyl butanoate and ethyl butanoate) and showed considerable variation between both manufacturers and batches. Flavouring VOCs are regulated for ingestion but are not regulated for e-liquid inhalation, so these could have toxicological implications. In addition, the inconsistency between samples suggests further issues when users add their own nicotine to the e-liquids as the viscous sample matrix makes homogeneous mixing difficult.

"They're sleek, stylish and sexy:" selling e-cigarettes online.

OBJECTIVE: We examined the product range, marketing strategies, access and marketing claims made by Australian and New Zealand (NZ) online e-cigarette retailers. METHODS: Twenty Australian (n=10) and NZ (n=10) e-cigarette retail websites were identified via Google using a combination of keywords nominated by an expert panel and identified via a literature review: 'e-cigarette', 'e-cigs', 'vape', and 'vaping', combined with 'Australia', 'AU', 'New Zealand' and 'NZ' and then examined. RESULTS: Products were extensive (disposable, pod-based, reusable, replacement parts), 95% (n=19) offered 'Starter Kits,' flavoured e-liquid (n=1,032), most containing nicotine (70%, n=14). Most retailers (85%, n=17) offered price discounts and free delivery. There were unsubstantiated health claims (80%, n=16), cessation claims (65%, n=13) and cost-benefit claims (50%, n=10) promoting e-cigarette use. Most (n=14) website age verification features simply required the purchaser to indicate they were aged 18 years. CONCLUSIONS: Although e-cigarette regulations are different in Australia and NZ, the online product range, marketing strategies, access and marketing claims were similar and sold e-liquid containing nicotine. The health and cessation e-cigarette marketing claims were outlandish and unsubstantiated. IMPLICATIONS FOR PUBLIC HEALTH: Most purchasing of e-cigarettes occurs online. Regulations and enforcement to limit access and stop unsubstantiated marketing claims must be a public health priority.

High content analysis of in vitro alveolar macrophage responses can provide mechanistic insight for inhaled product safety assessment.

Assessing the safety of inhaled substances in the alveolar region of the lung requires an understanding of how the respired material interacts with both physical and immunological barriers. Human alveolar-like macrophages in vitro provide a platform to assess the immunological response in the airways and may better inform the understanding of a response to an inhaled challenge being adaptive or adverse. The aim of this study was to determine if a morphometric phenotyping approach could discriminate between different inhaled nicotine products and indicate the potential mechanism of toxicity of a substance. Cigarette smoke (CS) and e-liquids extracted into cell culture medium were applied to human alveolar-like macrophages in mono-culture (ImmuONE™) and co-culture (ImmuLUNG™) to test the hypothesis. Phenotype profiling of cell responses was highly reproducible and clearly distinguished the different responses to CS and e-liquids. Whilst the phenotypes of untreated macrophages were similar regardless of culture condition, macrophages cultured in the presence of epithelial cells were more sensitive to CS-induced changes related to cell size and vacuolation processes. This technique demonstrated phenotypical observations typical for CS exposure and indicative of the established mechanisms of toxicity. The technique provides a rapid screening approach to determine detailed immunological responses in the airways which can be linked to potentially adverse pathways and support inhalation safety assessment.

Health effects and known pathology associated with the use of E-cigarettes.

In recent years, new nicotine delivery methods have emerged, and many users are choosing electronic cigarettes (e-cigarettes) over traditional tobacco cigarettes. E-cigarette use is very popular among adolescents, with more than 3.5 million currently using these products in the US. Despite the increased prevalence of e-cigarette use, there is limited knowledge regarding the health impact of e-cigarettes on the general population. Based on published findings by others, E-cigarette is associated with lung injury outbreak, which increased health and safety concerns related to consuming this product. Different components of e-cigarettes, including food-safe liquid solvents and flavorings, can cause health issues related to pneumonia, pulmonary injury, and bronchiolitis. In addition, e-cigarettes contain alarmingly high levels of carcinogens and toxicants that may have long-lasting effects on other organ systems, including the development of neurological manifestations, lung cancer, cardiovascular disorders, and tooth decay. Despite the well- documented potential for harm, e-cigarettes do not appear to increase susceptibility to SARS-CoV- 2 infection. Furthermore, some studies have found that e-cigarette users experience improvements in lung health and minimal adverse effects. Therefore, more studies are needed to provide a definitive conclusion on the long-term safety of e-cigarettes. The purpose of this review is to inform the readers about the possible health-risks associated with the use of e-cigarettes, especially among the group of young and young-adults, from a molecular biology point of view.

Expanding the E-Liquid Flavor Wheel: Classification of Emerging E-Liquid Flavors in Online Vape Shops.

INTRODUCTION: Electronic cigarettes are the most popular tobacco product among U.S. youth, and over 80% of current youth users of e-cigarettes use flavored e-cigarettes, with fruit, mint/menthol, and candy/sweets being the most popular flavors. A number of new e-liquid flavors are currently emerging in the online e-cigarette market. Menthol and other flavored e-cigarettes could incentivize combustible tobacco smokers to transition to e-cigarette use. METHODS: From February to May 2021, we scraped data of over 14,000 e-liquid products, including detailed descriptions of their flavors, from five national online vape shops. Building upon the existing e-liquid flavor wheel, we expanded the semantic databases (i.e., key terms) to identify flavors using WordNet-a major database for keyword matching and group discussion. Using the enriched databases, we classified 14,000+ e-liquid products into the following 11 main flavor categories: "fruit", "dessert/candy/sweets", "coffee/tea", "alcohol", "other beverages", "tobacco", "mint/menthol", "nuts", "spices/pepper", "other flavors", and "unspecified flavor". RESULTS: We find that the most prominent flavor sold in the five online vape shop in 2021 was fruit flavored products, followed by dessert/candy/other sweets. Online vendors often label a product with several flavor profiles, such as fruit and menthol. CONCLUSIONS: Given that online stores market products with multiple flavor profiles and most of their products contain fruit flavor, the FDA may have issued marketing denial orders to some of these products. It is important to further examine how online stores respond to the FDA flavor restrictions (e.g., compliance or non-compliance).

Histopathological scoring system role in evaluation of electronic cigarette's impact on respiratory pathway in albino rat: Biochemical, histo-morphometric and ultrastructural study.

BACKGROUND: Electronic cigarettes have gained growing popularity especially among adolescents and young adults. Although the dominant consideration by consumers and the companies claims, whether electronic smoking is indeed a much safer alternative to traditional smoking is hotly debated. AIM: To declare the biochemical, histopathological and ultrastructural impact of electronic cigarette exposure on the tissues of respiratory passage. MATERIAL AND METHODS: Twenty adult male Wistar albino rats were allocated into two groups of 10 animals in each; group I as control group and group II which was exposed to 1 ml/day for one hour of E-liquid involving (18 mg/ml nicotine) for 5 sequential days/ week for 4 weeks. Assessment of markers of oxidative stress; malondialdehyde (MDA) & total antioxidant capacity (TAC) was performed. Specimens of their larynxes and lungs were subjected to histopathological examination with light microscopy (H&E, toluidine blue and orcein stain) with histopathological injury scoring, electron microscopic, histo-morphometric, and immune-histochemical studies of protein α-SMA, p53 and ki-67 RESULTS: E-liquid vapors exposed rats exhibited a significant elevation of MDA and suppression in TAC levels in comparison with the control group in all respiratory pathway organs. Microscopic analysis of respiratory epithelium of exposed group revealed hyperplasia, loss of their cilia and vascular dilatation in lamina propria while the lung examination showed thickened alveolar septa, inflammatory infiltrate and congested thick wall blood vessels. The bronchus was lined with multilayered dark stained nuclei epithelium, and there were thickened irregular smooth muscle fibers. Morphometric analysis revealed increase of area percent of elastic fibers in experimental rats' lungs. Increase of α-SMA and P53 immuno-expression but decrease of Ki-67 immuno-expression in exposed groups were observed when compared to control. Ultrastructural examination showed cilial loss in respiratory epithelium with shrunken type 1 & 2 alveolar cells, with cytoplasmic vacuolations, atypical lamellar bodies and shrunken nuclei. Significant increase of the score of histopathological injury in larynx and lung. CONCLUSION: Exposure to e-liquid with nicotine induces oxidative stress and has adverse impact on respiratory pathway including histo-pathological and ultrastructural disorganization of respiratory epithelium and lung tissues.