September 14, 2022

Research Track Session
Thursday 14:00 - 14:30


Chairs: Konstantinos Poulas & Amaliya Amaliya

  • Toxicology and aerosol chemistry [31-34]

The fate of nicotine molecule, when heated at different temperatures in different surroundings (i.e. atmospheres) is of fundamental importance to harm reduction when inhaling the aerosols of different tobacco products. In the present study, we tackle this issue from the viewpoint of molecular modelling. Series of ab initio molecular dynamics simulations of free nicotine molecule as well as of nicotine molecule in the presence of oxygen molecules at series of different temperatures ranging from 600 to 1200 K are performed with the atom-centered density matrix propagation scheme (ADMP). Simulations are performed at different levels of theory. The possible degradation pathways implied by the generated molecular dynamics trajectories are related to the products detected in the smoke of conventional cigarettes and aerosols generated by electronic devices for controlled heating of tobacco.


AUTHORS: Mereme Idrizi1, Blagoj Achevski1, Hrisanta Godzo1, Jasmina Tonic-Ribarska1, Ljupcho Pejov2,3

AFFILIATIONS: 1Faculty of Pharmacy, SS. Cyril and Methodius University, Skopje, North Macedonia || 2Faculty of Natural Sciences and Mathematics, SS. Cyril and Methodius University, Skopje, North Macedonia || Department of Chemistry, Bioscience and Environmental Engineering, Faculty of Science and Technology, University of Stavanger, Norway

Background: Heated tobacco products (HTPs) are becoming more and more popular during the last decades among smokers. HTPs recent innovative non-combusted products due to heating of tobacco up to 350°C (lower than 600°C as in conventional cigarettes). Unfortunately, combustible cigarette smoking is the most important preventable cause of cancer. Therefore, strict standards and requirements have been developed and applied to these products. Thus, HTPs offer alternatives to combustible cigarettes and have partially replaced combustible cigarettes. Moreover, the HTPs use is rising, but potential health risk in the result of long-term use has not been studied completely. The aim of the study is to analyze and compare the different methodologies based on evaluation of the cancer risk associated with the use of HTPs.

Material and Methods: Advanced bibliographic study of 35 bibliographic sources from databases: Medline, Scopus, HINARI, PubMed, and Cochrane Electronic.

Results: The cancer risk assessment using different methodologies were analyzed and compared. The first method was based on the change in cumulative exposure (CCE) of the compounds emitted. Another was focused on margin of exposure (MOE) determination, which represented as the ratio between the toxicological thresholds of the compound to the estimated human expose (with a higher MOE indicating a lower risk). The next one attempted to estimate the cancer potencies of various product types using levels of chemical constituents found in the aerosols of each product type and their associated inhalation unit risks.

Conclusions: Three modelling cancer risk assessments estimated that the heated tobacco product under performed investigations had a lower cancer potency (compared to that of conventional cigarettes).


AUTHOR: Ecaterina Mazur

AFFILIATION: Nicolae Testemitanu University State of Medicine and Pharmacy, Chisinau, Republic of Moldova

E-cigarette devices that youth prefer have evolved substantially over time, from early-generation cigar like ECs, to more advanced modifiable tank-style ECs, to the recent emergence of a sophisticated, sleekly designed, discreet high-tech device called JUUL. JUUL is an electronic cigarette which uses nicotine salts, prefilled with a high concentration in pods, rather than free-base nicotine. Due to its popularity among young ages, concerns have been voiced and public health officials have pressed the FDA to step in. E-liquids contain various combinations of nicotine, flavors, and carriers. These factors affect nicotine delivery, appeal, and ease of product use and underscore the degree to which individual preferences may play a role in use patterns. Cartridge nicotine solution in ECs seems to exacerbate asthmatic symptoms by elevating infiltration of inflammatory cells including eosinophils into airways and promotion of oxidative stress.

The aim of this study is to evaluate the effects of JUUL on human alveolar cell lines of upper respiratory and bronchial epithelium, besides with conventional cigarette exposure, on cell toxicity and oxidative stress. AqE extracts from JUUL different flavors were created in different concentrations with Borgwaldt LM4E vaping machine, to determine the concentration that exhibits toxicity. 1R6F reference cigarette was used as control and the condensates prepared with Borgwaldt LM1 smoking machine. A549 human alveolar cell line and NCI-H292 bronchial epithelial cells were tested. Cell toxicity was monitored with MTT assay for cell viability and Superoxide Dismutase Assay was performed as indicator of ROS activity. The results indicate a loss of viability only in high puff volumes, in high concentrated AqEs, with small differences between different flavors in 24h treatment. In 48h treatment differences were observed between flavors toxicity. SOD induction observed in different flavors between the two cell lines.


AUTHORS: Konstantinos Mesiakaris1,2, Aggeliki Thanasoula1, Maria C. Kotsira1, Marilena Kaperoni1,2, Konstantinos Poulas1,2

AFFILIATIONS: 1Laboratory of Molecular Biology and Immunology, Department of Pharmacy, University of Patras, Rio, Greece || 2Institute for Research and Innovation, IRIS, Patras Science Park, Patras, Greece

There are several different methods that have been used to investigate and assess the biological effects of cigarette smoke, with a range at the design of an inhalation machine. Smoking Machines are designed to fulfill the requirements of in vitro experiments as well as of various other research tasks. Μany old studies used hand-made smoking delivery machines and it was difficult to transfer the knowledge and maintain strict conditions on the experiments. Newest studies use experimental set-ups with automated methods, that can follow experimental regimes such as Health Canada Intense (HCI), ISO and CRM and control the experimental parameters strictly, such as puff volume, puff duration, puff profile etc. Several in vitro techniques have been reported, which involve the exposure of submerged cellular cultures to either total particulate matter (TPM) or aqueous cigarette smoke extracts.

The aim of the present study was to set-up LM4E vaping and LM1 smoking machines by Borgwaldt and the use of Air liquid Interface (ALI) technique for experimentation. Cell cultures performed at the air-liquid interface facilitate the establishment of stable and functional 3D models of the respiratory tract. In these models, the basal side of the cells is in contact with the culture medium, and the apical side is in contact with the air. For that purpose, 1R6F cigarette was tested on NCI-H292 cells, in two ways, as whole smoke and vapor phase, following ISO and HCI regimes. Cell viability was measured using NRU assay.


AUTHORS: Konstantinos Mesiakaris1,2, Marilena Kaperoni1,2, Konstantinos Poulas1,2

AFFILIATIONS: 1Laboratory of Molecular Biology and Immunology, Department of Pharmacy, University of Patras, Rio, Greece || 2Institute for Research and Innovation, IRIS, Patras Science Park, Patras, Greece


  • Preclinical evaluation [35]

Among chemicals found in cigarette smoke, World Health Organization Study Group on Tobacco Product Regulation (WHO TobReg) identified six volatile aldehydes regarded as priority toxicants. An estimation of exposure to aldehyde from cigarette smoke and risk assessment studies show that due to their pervasiveness and documented toxicity, acrolein, acetaldehyde, formaldehyde, propionaldehyde, crotonaldehyde, and butyraldehyde have particularly detrimental effects on human health. While mandatory lowering aldehyde levels in cigarette smoke is a part of regulatory strategy for reducing exposure of smokers to toxicants, pharmacological approaches mitigating their health impact are also considered. For example, aldehyde-related damage can be mitigated by boosting endogenous aldehyde dehydrogenase (ALDH) activity. ALDH is a superfamily of 19 enzymes that catalyze oxidation of aldehydes to carboxylic acids. This enzymatic activity prevents the accumulation of aldehydes derived i.a. from exogenous exposures and represents a major cytoprotective mechanism of the ALDH family members.

A recent study revealed that β-escin, a mixture of triterpenoid saponins extracted from the seed of horse chestnut tree (Aesculus hippocastanum L), is a strong activator of ALDH activity. Here we report the effect of β-escin on ALDH activity and gene expression in primary human nasal epithelial cells exposed to cigarette smoke extract (CSE). We revealed that while both β-escin (1μM) and CSE (5%) stimulated ALDH activity, their combination evoked much more pronounced response (at 24h: 132% and 137% vs 174%, respectively). The observed increase in the enzyme activity correlated with an increased expression of ALDH1A3, ALDH3A2, ALDH3B1 and ALDH18A and was significantly higher in the cells treated with the combination of β-escin and CSE. Importantly, β-escin reversed CSE-induced inhibition of ALDH1A1 and ALDH2 expression.

These results suggest that increasing the ALDH gene expression and enzyme activity by β-escin may facilitate aldehyde removal and thus be useful in reducing cigarette smoke toxicity. Such approach might be a part of harm reduction strategy which should be applied for smokers that are unwilling to quit or failed to quit smoking conventional cigarettes.


AUTHORS: Malwina Sołtysiak, Magdalena Paplińska-Goryca, Paulina Misiukiewicz-Stępień, Paulina Wójtowicz, Rafał Krenke, Katarzyna Koziak

AFFILIATION: Medical University of Warsaw, Poland


  • Biomarkers’ evaluation in animal or human studies [36]

Background: Electronic cigarettes (ECigs) use has increased substantially in recent years. ECigs have been proposed as a potentially effective smoking cessation tool, they deliver nicotine, thus dealing with the chemical part of the addiction, and, at the same time, they provide sensory and motor stimuli like smoking. In addition, the flavored liquid ECigs from plant extracts have been widely used as potentially beneficial due to plant properties.

The winemaking procedure results in the production of stems, by-products that are not environmentally friendly. However, grape stems are rich in polyphenols and, therefore, they are putatively beneficial for human health.

The aim of the present study is to investigate the effect of the grape stems extract on behavior, redox status (liver and brain regions) adult male mice and its possible use in e-liquids maintaining its antioxidant and anxiolytic properties.

Material and Methods: The grape stem extract was derived from a native Greek vine, namely Mavrodaphne and was rich in polyphenols (205.2 mg/g extract). LM4E Vaping Machine by Borgwaldt was used to create an aquatic extract enriched with vaping constituents of the E-liquid (VG/nicotine) enriched with grape stems aquatic extracts (AqE).

The stem extract was administrated in two different ways a) intraperitoneally (IP) (AqE that contains nicotine, VG, stem extract and saline) and b) orally (gavage) (stem extract) for 28 days (long-term). Behavior analysis was assessed by using the open-field test, followed by video-tracking software (Any-maze 6.3) analysis. Antioxidant analysis was performed by determining the SOD activity.

Conclusions: Anxiolytic-like behavior was observed after the stem extract and aqueous extract administration. Stem extract treatments also showed antioxidant effect. In conclusion, long term administration of grape stem extract, rich in polyphenols, appears to have anxiolytic, antioxidant activity.

AUTHORS: Korina Atsopardi1,2, Konstantinos Mesiakaris1, Dimitrios E. Providas1, Panagiota Giannakopoulou3, Marigoula Margarity2, Konstantinos Poulas1

AFFILIATIONS: 1Laboratory of Molecular Biology and Immunology, Department of Pharmacy, University of Patras, Rio, Greece || 2Laboratory of Human and Animal Physiology, Department of Biology, University of Patras, Rio, Greece || 3Laboratory of Developmental Biology, Department of Biology, University of Patras, Rio, Greece

Poster presentations: 5 min