Background: Cisplatin resistance represents a major barrier to effective treatment of head and neck squamous cell carcinoma (HNSCC). A previous study by Manyanga et al. (2021) reported that nicotine and e-cigarette (e-cig) aerosols may enhance cisplatin resistance in oral cancer models. This international replication study aimed to assess those findings using harmonized protocols across multiple laboratories and to investigate the potential impact of e-cig aerosols on cisplatin responsiveness in HNSCC.

Material and Methods: Standardized in vitro experiments were conducted using three HNSCC cell lines (SCC-25, FaDu, UM-SCC-1). Cells were exposed to aqueous extracts of cigarette smoke (1R6F) and e-cig aerosols with 0, 12, or 20 mg/mL nicotine. Extracts were generated in PBS using ISO-compliant smoking/vaping machines (ISO20778:2018; ISO20768:2018) and diluted to obtain a 10 puffs/5L concentration. Cytotoxicity (MTS, NRU, trypan blue), clonogenic survival, and gene/protein expression (qRT-PCR, Western blot) were assessed to evaluate the influence of aqueous extracts on cisplatin effectiveness.

Results: Differently from the original work, our results showed no consistent alteration in cisplatin sensitivity following e-cig exposure. IC50 values, cell viability, and colony-forming ability were not significantly different from cisplatin controls. Although some variations in DNA repair gene and transporter protein expression were detected, these were not reproducible across models or conditions.

Conclusions: Our data do not support a generalized link between e-cigarette exposure and increased cisplatin resistance in HNSCC. The variability observed highlights potential context-specific effects and underscores the need for rigorous cross-laboratory validation before drawing clinical conclusions.

AUTHORS: Giuseppe Carota¹, Rosalia Emma^2,3, Konstantinos Partsinevelos¹, Sonja Rust³, Ang Sun⁴, Antonio Giordano^4,5,6, Vladislav Volarevic^7,8,9, Ronny Lesmana^10,1112, Hanna Goenawan^10,11,12, Melisa Intan Barliana¹³, Aleksandar Arsenijevic^7,8,9, Nikolina Kastratovic^7,8, Bogdan Spasic⁸, Chiara Giardina¹, Miriana Cantali¹, Alessandra Pittalà¹, Miriam Wissam Saab¹, Angela Maria Amorini¹, Riccardo Polosa^14,15, Giovanni Li Volti^1,15, Massimo Caruso^1,15

AFFILIATIONS: ¹Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy

²Department of Medicine and Surgery, “Kore” University of Enna, Contrada Santa Panasia, Enna, Italy

³ECLAT Srl, spin off of the University of Catania, Catania, Italy

⁴Department of Biology, College of Science and Technology, Temple University, Philadelphia, PA 19122, USA

⁵Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, College of Science and Technology, Temple University, Philadelphia, PA 19122, USA

⁶Department of Medical Biotechnologies, University of Siena, Italy

⁷Center for harm reduction of biological and chemical hazards, Faculty of Medical Sciences University of Kragujevac, Kragujevac, Serbia

⁸Department of Genetics, Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia

⁹Department of Microbiology and Immunology, Faculty of Medical Sciences, University of Kragujevac, Kragujevac, Serbia

¹⁰Department of Biomedical Sciences, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia

¹¹Division of Biological Activity, Central Laboratory, Universitas Padjadjaran, Bandung, Indonesia

¹²Center of Excellence for Pharmaceutical Care Innovation, Universitas Padjadjaran, Bandung, Indonesia

¹³Department of Biological Pharmacy, Faculty of Pharmacy, Universitas Padjadjaran, Bandung, Indonesia

¹⁴Department of Clinical and Experimental Medicine, University of Catania, Catania, Italy

¹⁵Center of Excellence for the Acceleration of Harm Reduction (CoEHAR), University of Catania, Catania, Italy

Background: Tobacco smoking is the leading cause of preventable morbidity and mortality worldwide. In recent years, modified risk products (MRPs), including heated tobacco products (HTPs), have transformed the tobacco industry, attracting a growing number of users. This study investigates the impact of combustible cigarette smoke and HTP aerosol on microglial function, with particular focus on the metabolomic profile, under ISO standard and clinically relevant conditions of exposure.

Material and Methods: Human microglial cells (HMC3) were exposed to aqueous extract (AqE) of cigarette smoke (1R6F reference cigarette, University of Kentucky), AqE of aerosol from a commercially available HTP (IQOS 3 DUO, Philip Morris International) and nicotine, each at the same nicotine concentration (340 nM), which falls within the range detected in the brain of smokers, in order to compare metabolomic effects. Smoke and aerosol AqEs were prepared using ISO standard procedures. Nicotine identification and quantification in AqEs, as well as evaluation of the metabolomic profile in exposed cells, were performed using a protocol suitable for HPLC analysis.

Results: Our findings show that cigarette smoke significantly reduces GSH levels and increases NADP+/NADPH ratio, indicating a shift toward oxidative stress not observed with IQOS or nicotine. Additionally, cigarette smoke also alters the intracellular energy charge and decreases total triphosphates, reflecting a suffering state compared to all other treatments. Evidence of mitochondrial dysfunction induced by cigarette smoke is further supported by a remarkable decrease in the ATP/ADP ratio, whereas only a slight alteration was detected after IQOS exposure. Furthermore, NAD+/NADH ratio, a key parameter for oxidative phosphorylation efficiency, decreased only with cigarette smoke.

Conclusions: These results highlight the different effects of combustible cigarette and HTP on the metabolomic profile of microglial cells, suggesting a potential harm reduction strategy in the context of neurodegenerative diseases for smokers unwilling or unable to quit.

AUTHORS: Konstantinos Partsinevelos¹, Alfio Distefano¹, Laura Orlando¹, Lucia Longhitano¹, Rosalia Emma^2,3,4, Massimo Caruso^1,3,5, Nunzio Vicario¹, Simona Denaro¹, Ang Sun^5,6, Antonio Giordano^5,6, Barbara Tomasello⁷, Amer M. Alanazi⁸, Giovanni Li Volti^1,3, Angela Maria Amorini¹

AFFILIATIONS: ¹Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, Catania (CT), Italy

²Department of Clinical and Experimental Medicine, University of Catania, Catania (CT), Italy

³Center of Excellence for the Acceleration of Harm Reduction (CoEHAR), University of Catania, Catania (CT), Italy

⁴Department of Medicine and Surgery, "Kore" University of Enna, Contrada Santa Panasia, Enna, Italy

⁵Department of Biology, College of Science and Technology, Sbarro Institute for Cancer Research and Molecular Medicine, Temple University, Philadelphia, USA

⁶Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, College of Science and Technology, Temple University, Philadelphia, USA

⁷Department of Drug and Health Science, Section of Biochemistry, University of Catania, Catania (CT), Italy

⁸Pharmaceutical Biotechnology Laboratory, Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia

Background: Smoking status (SS) is associated with structural and functional vascular changes which may lead to increased arterial stiffness and, as a consequence, the formation of early vascular aging.

The purpose of this study was to investigate the relationship between the modifiable risk factors and early vascular aging (EVA-syndrome).

Material and Methods: We analyzed data from 783 men aged 20-92 (mean age 48.2 ± 10.32 years) undergoing medical examinations. Vascular age (VA) was assessed using photoplethysmography (AngioScan-01), evaluating the stiffness index (SI), reflection index (RI), augmentation index (AIp75), index (AGI). EVA syndrome was defined as AGI values above the 90th percentile, while young VA was defined as AGI values below the 10th percentile. Correlations with passport age were analyzed.

Results: The “age index” (AGI) showed the highest correlation with passport age (r=0.54, p<0.000). EVA-syndrome criteria were considered as values of the AGI, beyond the 90th percentile (n = 65), criteria for “young vascular age” (n=87) were AGI values below 10th percentile. Comparison of these two alternative subgroups (in men aged from 40 to 59) in terms of arterial stiffness showed significant differences in terms of SI, Alp75, PV-type. The leading vascular risk factors for EVA syndrome were arterial hypertension (OR=4.4; p<0.05), hyperglycemia (OR=3.2; p<0.05), hypertriglyceridemia (OR) = 2.9; p< 0.05), smoking status (OR = 2.23, p<0.05) Long-term (more than 20 years) smoking history was associated with 11.6 potential years of life lost compared with never smokers, former smokers, or users of reduced-risk products.

Conclusions: Our findings emphasize the importance of developing algorithms for clinicians using the harm reduction concept in the fight against smoking as a key behavioral risk factor for the development of ASCVD and a reduction in life expectancy and recommendations for the use of less harmful alternatives to slow or reverse the progression of arterial ageing.

AUTHORS: Aleksander Filippov, Artur Tuktarov

AFFILIATION: St. Petersburg State University, St Petersburg, Russian Federation

Background: The gradual shift from the widespread use of traditional cigarettes to the active use of alternative nicotine delivery systems (ANDS) in today’s young population aged 35-44 years determines the need for scientific research aimed at objectifying their impact on the body.

The objective of this study was to investigate hemodynamic, metabolism, proinflammatory indices in healthy men depending on the nicotine-containing products consumption ways.

Material and Methods: The cohort (n=411) was divided into three groups comparable in age and body mass index according to smoking status: non-smoking subjects (NS) (n=275; age 40.94±4.90); traditional cigarette smokers (TCSs) (n=68; age 41.09±3.85); ANDS users (n=68; age 39.82±6.31). Data of office blood pressure (BP) and heart rate (HR), blood biochemical analysis were studied. There was used nonparametric statistics (Mann-Whitney U test).

Results: The systolic BP was significantly higher in TCSs and АNDS users than in NS (p=0,02, p=0,04 respectively), HR in TCSs was significantly higher than in NS (p=0,004). High-density lipoproteins (HDL) levels were the lowest and triglyceride (TG), C-reactive protein (CRP), fibrinogen levels were the highest in TCSs. HDL levels didn’t differ between NS and АNDS users (p>0.05). CRP and fibrinogen levels were higher in TCSs than in NS (p=0.08, p=0.56 respectively) and АNDS users (p=0.003, p=0.62 respectively).

Conclusions: Analysis of cardiovascular risk biomarkers such as HDL, TG, CRP, and fibrinogen levels revealed a positive trend in the АNDS users compared with TCSs, indicating a reduction in risk. The concept of harm reduction realized by ‘switching’ from cigarette smoking to АNDS use may improve the cardiovascular prognosis. Further short- and long-term АNDS studies are needed to assess their impact on smoker’s health compared with smoking traditional cigarettes and to explore the potential for incorporating the results of these studies into recommendations for the prevention and treatment of tobacco-associated diseases.

AUTHORS: Anton Barsukov¹, Vladislav Dydyshko², Ekaterina Borisova¹, Svetlana Glebova¹

AFFILIATIONS: ¹KardioKlinika, St. Petersburg, Russian Federation

²S.M. Kirov Military Medical Academy, St. Petersburg, Russian Federation

Background: This study aimed to evaluate the association between traditional smoking, vaping, BMI, and the incidence of anastomotic leak in patients undergoing colorectal surgery. A key objective was to identify vaping as an independent variable associated with anastomotic leak and wound healing in this population.

Material and Methods: A retrospective cohort study was conducted to assess these associations. Multivariable analysis was performed, adjusting for patient comorbidities and surgical approach, to determine independent risk factors for various complications.

Results: After adjustments, traditional smoking was identified as an independent risk factor for wound complications (OR 1.31, 95% CI 1.02-1.70, p = 0.035), primary pulmonary complications (OR 1.40, 95% CI 1.08-2.06, p = 0.021), and anastomotic leak (OR 1.47, 95% CI 1.20-2.46, p = 0.003). While major medical complications, surgical complications, and sepsis showed a trend towards higher risk in smokers, these did not reach statistical significance. Importantly, no significant association was found between vaping and anastomotic leak.

Conclusions: Our findings demonstrate that traditional smoking is an independent risk factor for anastomotic leak and other complications in colorectal surgery patients. These results underscore the critical need for preoperative smoking cessation strategies. Conversely, we found no evidence linking vaping to this complication. Surgeons should emphasize these elevated risks to patients considering elective colorectal surgery.

AUTHOR: Fernando Fernandez Bueno

AFFILIATION: Plataforma de Reducción de Daños por Tabaquismo, Spain

Background: Heated tobacco products (HTPs) are marketed as reduced-risk alternatives to traditional tobacco cigarettes (TCs). However, the actual reduction in toxicological burden and health benefits remains debated. We evaluated preclinical and clinical studies comparing HTPs to TCs, focusing on aerosol composition, toxicological exposure, and biomarkers of harm.

Material and Methods: A systematic PubMed search identified 1,105 peer-reviewed studies published between January 2017 and October 2024. We selected 45 studies reporting standardized aerosol characterization, in vivo/in vitro toxicology, or biomarker analysis in humans.

Results: HTPs heat rather than burn tobacco, producing aerosol with significantly lower (up to 95%) toxicant levels and reduced toxic, mutagenic, genotoxic, carcinogenic, and proinflammatory potentials by 85–95% compared to TC smoke in vitro and in vivo. Among 26 human interventional studies, 22 reported 40–97% reductions in toxicant biomarkers of exposure (BoE) —such as tobacco-specific nitrosamines, carboxyhemoglobin, volatile organic compounds, and mutagenic metabolites— in participants who switched from TCs to HTPs. These effects were observed across various time frames, ranging from minutes to 24 months. Four independent studies found no significant improvement or identified adverse effects. Most BoE changes occurred independently of nicotine blood levels. International health authorities confirmed that HTP aerosol contains some mutagenic and carcinogenic compounds, but at much lower levels than TC smoke, thus reducing health risk.

Conclusions: HTPs reduce toxicant exposure relative to TCs, but further independent, long-term studies are needed to determine their full potential health impact.

AUTHORS: Stefano Bellosta¹, Alberto Corsini¹, Emanuele Catena², Gabriele Catena²

AFFILIATIONS: ¹University of Milan, Milan, Italy

²COO Sismed Servizi srl, Rome, Italy

Background: Tobacco-free nicotine pouches (TFNPs) do not contain tobacco and the levels of toxic chemicals in their extracts are like those in medicinal nicotine products such as nicotine gums and lozenges, which means that they are likely to be the least harmful of any currently available tobacco/nicotine product and to occupy a position to the extreme right of the risk continuum. The value of any reduced-risk products like TFNPs, however, lies in smokers’ willingness to use them instead of cigarettes. Here we seek to understand the choices consumers make as they move down the risk continuum from combustible cigarettes at the extreme left to TFNPs at the extreme right.

Material and Methods: We conducted an online survey of 1,409 TFNP users in the US, which was designed to understand tobacco and nicotine product use pathways.

Results: Transitions down the risk continuum among TFNP users involved multiple intermediate steps before arriving at use of lower-risk products. Starting to smoke cigarettes was the most common first-used tobacco product. TFNPs were not the most common second product used. Most cigarette smokers tended to transition to dipping/chewing tobacco and vapour products, with some transitioning to using snus. At further transitions however, these were predominantly to TFNPs, although the ultimate path to TFNP use occurred as far as the sixth transition. Transitioning to TFNP use, regardless of chronology and across all transitions, was most commonly from dipping/chewing tobacco, slightly less commonly from vapour products, and much less commonly from cigarettes. Transitioning from TFNPs to cigarettes was very rare (1.2% of all transitions).

Conclusions: These findings will help the understanding of the population-level impact of TFNPs and of the contribution they can make to Tobacco Harm Reduction. The findings may be used to tailor interventions to maximise THR by promoting more rapid transit down the risk continuum.

AUTHOR: Marina Murphy

AFFILIATION: Senior Director Scientific Affairs, Haypp Group, London, England

Background: Pakistan faces severe health threats from tobacco smoking and indoor/outdoor air pollution, leading to rising respiratory and cardiovascular diseases. This project aimed to reduce these risks by training Lady Health Workers (LHWs) as advocates for healthy lung practices, focusing on smoking cessation and clean air.

Material and Methods: In collaboration with District Health Authorities (DHA) and Basic Health Units (BHUs), 46 LHWs were recruited from four primary health units across three Tehsils. They disseminated HLFL messages to (7,360 households (47,840 individuals) through: awareness pamphlets on avoiding polluted air and smoking risks; health education campaigns integrating HLFL into routine LHW activities; and advocacy on vaccination and physical activity.

Table: HLFL Key Interventions

*a district administration or revenue subdivision

Results:

• LHWs successfully educated households on reducing indoor pollution (e.g., avoiding active & passive smoking).
• Increased awareness of Sudden Infant Death Syndrome (SIDS) and respiratory risks from second-hand smoke.
• Strengthened preventive healthcare through health education promotion.

Conclusions: The HLFL initiative effectively leveraged LHWs to combat respiratory health risks in rural Pakistan. Sustained efforts in policy enforcement, smoke-free environments, and community education are crucial for long-term impact.

AUTHOR: Qamar Iqbal

AFFILIATIONS: European Lung Foundation (ELF) & European Respiratory Society (ERS), Gujranwala, Pakistan

Background: In recent years, a number of publications have discussed the potential role of e-cigarettes (ECs) and heated tobacco products (HTPs) in smoking cessation.

Material and Methods / Results: The comprehensive review from the Cochrane Library, updated in January 2025, again confirmed that ECs with nicotine increase quit rates compared to nicotine-replacement therapy with high-certainty evidence (RR 1.59, 95% CI 1.30–1.93).

In the Korean nationwide study (2025) of smokers with coronary artery disease undergoing percutaneous coronary intervention, switching to ECs was associated with a significantly lower risk of major adverse cardiac events than continued use of combustible cigarette, similar to quitting smoking (HR 0.82, 95% CI 0.69–0.98 for switchers to ECs and HR 0.87, 95% CI 0.79–0.96 for successful quitters).

A recent scoping review of behavioral studies of HTPs (2024) found that positive individual‑level benefits of transitioning to the use of HTPs may also be seen at the population level.

As the 2023 AHA/ACC Guideline for the Management of Patients with Chronic Coronary Disease, the 2024 ESC Guidelines for the management of peripheral arterial and aortic diseases classified the ECs as a possible smoking cessation tool with a Class IIb recommendation. These guidelines emphasize that ECs use is associated with adverse effects on cardiovascular and other systems compared with non-users, but with milder effects than smoked cigarettes. The 2024 ESC Guidelines for the management of chronic coronary syndromes indicate that the use of ECs should only be considered as an adjunct to a formal tobacco cessation program.

Conclusions: While ECs may be considered as a cessation tool for certain patients, significant concerns remain regarding their long-term safety and the potential for prolonged nicotine dependence. The overall consensus of leading cardiology societies is that complete abstinence from all nicotine products remains the optimal goal for cardiovascular health.

AUTHORS: Daria Regushevskaia^1,2, Yulia Yufereva¹

AFFILIATIONS: ¹N.I. Pirogov Russian National Research Medical University, Moscow, Russian Federation

²City Clinical Hospital No. 1 n.a. N.I. Pirogov, Moscow, Russian Federation

Background: Most patients who have suffered from an acute coronary syndrome (ACS) temporarily stop smoking for a short period after the event. Unfortunately, most of them start smoking again within three months. The study aims to assess the factors that influence smoking behavior after an ACS.

Material and Methods: The study has included 420 patients (320 male and 100 female, aged 58.2±14.3 years). All patients were active and heavy smokers (>20 cigarettes/day) at admission. Smoking was the only risk factor in 128 male and 18 female patients. We collected patients’ medical histories, risk factors, and smoking habits from their medical records. We used a questionnaire to record additional information, such as nicotine dependence (Fagerstrom Test). During hospitalization, all patients stopped smoking, and we advised them to visit the smoking cessation clinic to receive specialized help. We followed up with all participants for one year after discharge.

Results: Out of 420 patients, 223 (53.1%) quit smoking within the first 3 months after the episode, and 258 (61.4%) within 12 months. Among those who continued to smoke, most patients (90.1%) reported that their nicotine consumption was lower after the episode than before. In multivariate analysis, independent factors for smoking cessation were: participation in the Clinic’s smoking cessation program (OR=4.53, p=0.0007), lack of family history of smoking (OR=2.54, p=0.001), younger age (OR=2.1, p=0.001), better educational level (OR=1.89, p=0.01), and female gender (OR=1.7, p=0.01). Independent factors for continuing smoking were antidepressant drug use (OR=2.28, p=0.01), history of vascular disease (OR=2.32, p=0.03), history of COPD (OR=1.35, p=0.001), and high level of nicotine dependence (score >8 on the Fagerstrom Test for Nicotine Dependence) (OR=1.42, p=0.04).

Conclusions: Although quitting smoking is self-evident after an ACS, many factors can influence this behavior. Anti-smoking campaigns, as well as smoking cessation clinics, can successfully contribute to the desired outcome.

AUTHORS: Evangelos Sdogkos, Martha Amoiradou, Angelos Georgakopoulos, Ioannis Vogiatzis

AFFILIATION: Smoking Cessation Clinic, Department of Cardiology, General Hospital of Veroia, Veroia, Greece

Withdrawn

AUTHORS: Hifni Alifahmi¹, Ilham Kariem²

AFFILIATIONS: ¹Sahid University, Jakarta, Indonesia

²KABAR – Indonesian Tar Free Coalition/Koalisi Indonesia Bebas Tar