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      Elevated Cellular Oxidative Stress in Circulating Immune Cells in Otherwise Healthy Young People Who Use Electronic Cigarettes in a Cross‐Sectional Single‐Center Study: Implications for Future Cardiovascular Risk

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          Abstract

          Background

          Tobacco cigarettes (TCs) increase oxidative stress and inflammation, both instigators of atherosclerotic cardiac disease. It is unknown if electronic cigarettes (ECs) also increase immune cell oxidative stress. We hypothesized an ordered, “dose‐response” relationship, with tobacco‐product type as “dose” (lowest in nonsmokers, intermediate in EC vapers, and highest in TC smokers), and the “response” being cellular oxidative stress (COS) in immune cell subtypes, in otherwise, healthy young people.

          Methods and Results

          Using flow cytometry and fluorescent probes, COS was determined in immune cell subtypes in 33 otherwise healthy young people: nonsmokers (n=12), EC vapers (n=12), and TC smokers (n=9). Study groups had similar baseline characteristics, including age, sex, race, and education level. A dose‐response increase in proinflammatory monocytes and lymphocytes, and their COS content among the 3 study groups was found: lowest in nonsmokers, intermediate in EC vapers, and highest in TC smokers. These findings were most striking in CD14 dimCD16 + and CD14 ++CD16 + proinflammatory monocytes and were reproduced with 2 independent fluorescent probes of COS.

          Conclusions

          These findings portend the development of premature cardiovascular disease in otherwise healthy young people who chronically vape ECs. On the other hand, that the COS is lower in EC vapers compared with TC smokers warrants additional investigation to determine if switching to ECs may form part of a harm‐reduction strategy.

          Registration

          URL: https://www.clinicaltrials.gov; Unique identifier: NCT03823885.

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          Most cited references69

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          Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling.

          Reactive oxygen species (ROS) are generated during mitochondrial oxidative metabolism as well as in cellular response to xenobiotics, cytokines, and bacterial invasion. Oxidative stress refers to the imbalance due to excess ROS or oxidants over the capability of the cell to mount an effective antioxidant response. Oxidative stress results in macromolecular damage and is implicated in various disease states such as atherosclerosis, diabetes, cancer, neurodegeneration, and aging. Paradoxically, accumulating evidence indicates that ROS also serve as critical signaling molecules in cell proliferation and survival. While there is a large body of research demonstrating the general effect of oxidative stress on signaling pathways, less is known about the initial and direct regulation of signaling molecules by ROS, or what we term the "oxidative interface." Cellular ROS sensing and metabolism are tightly regulated by a variety of proteins involved in the redox (reduction/oxidation) mechanism. This review focuses on the molecular mechanisms through which ROS directly interact with critical signaling molecules to initiate signaling in a broad variety of cellular processes, such as proliferation and survival (MAP kinases, PI3 kinase, PTEN, and protein tyrosine phosphatases), ROS homeostasis and antioxidant gene regulation (thioredoxin, peroxiredoxin, Ref-1, and Nrf-2), mitochondrial oxidative stress, apoptosis, and aging (p66Shc), iron homeostasis through iron-sulfur cluster proteins (IRE-IRP), and ATM-regulated DNA damage response. Copyright © 2012 Elsevier Inc. All rights reserved.
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            Do multiple outcome measures require p-value adjustment?

            Background Readers may question the interpretation of findings in clinical trials when multiple outcome measures are used without adjustment of the p-value. This question arises because of the increased risk of Type I errors (findings of false "significance") when multiple simultaneous hypotheses are tested at set p-values. The primary aim of this study was to estimate the need to make appropriate p-value adjustments in clinical trials to compensate for a possible increased risk in committing Type I errors when multiple outcome measures are used. Discussion The classicists believe that the chance of finding at least one test statistically significant due to chance and incorrectly declaring a difference increases as the number of comparisons increases. The rationalists have the following objections to that theory: 1) P-value adjustments are calculated based on how many tests are to be considered, and that number has been defined arbitrarily and variably; 2) P-value adjustments reduce the chance of making type I errors, but they increase the chance of making type II errors or needing to increase the sample size. Summary Readers should balance a study's statistical significance with the magnitude of effect, the quality of the study and with findings from other studies. Researchers facing multiple outcome measures might want to either select a primary outcome measure or use a global assessment measure, rather than adjusting the p-value.
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              The pathophysiology of cigarette smoking and cardiovascular disease: an update.

              Cigarette smoking (CS) continues to be a major health hazard, and it contributes significantly to cardiovascular morbidity and mortality. Cigarette smoking impacts all phases of atherosclerosis from endothelial dysfunction to acute clinical events, the latter being largely thrombotic. Both active and passive (environmental) cigarette smoke exposure predispose to cardiovascular events. Whether there is a distinct direct dose-dependent correlation between cigarette smoke exposure and risk is debatable, as some recent experimental clinical studies have shown a non-linear relation to cigarette smoke exposure. The exact toxic components of cigarette smoke and the mechanisms involved in CS-related cardiovascular dysfunction are largely unknown, but CS increases inflammation, thrombosis, and oxidation of low-density lipoprotein cholesterol. Recent experimental and clinical data support the hypothesis that cigarette smoke exposure increases oxidative stress as a potential mechanism for initiating cardiovascular dysfunction.
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                Author and article information

                Contributors
                hmiddlekauff@mednet.ucla.edu
                Journal
                J Am Heart Assoc
                J Am Heart Assoc
                10.1002/(ISSN)2047-9980
                JAH3
                ahaoa
                Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease
                John Wiley and Sons Inc. (Hoboken )
                2047-9980
                08 September 2020
                15 September 2020
                : 9
                : 18 ( doiID: 10.1002/jah3.v9.18 )
                : e016983
                Affiliations
                [ 1 ] Division of Infectious Disease Department of Medicine David Geffen School of Medicine at UCLA Los Angeles CA
                [ 2 ] Division of Cardiology Department of Medicine David Geffen School of Medicine at UCLA Los Angeles CA
                [ 3 ] Department of Medicine David Geffen School of Medicine at UCLA Los Angeles CA
                [ 4 ] Department of Computational Medicine David Geffen School of Medicine at UCLA Los Angeles CA
                Author notes
                [*] [* ] Correspondence to: Holly R. Middlekauff, MD, Division of Cardiology, Department of Medicine, David Geffen School of Medicine at UCLA, A2-237 CHS, 650 Charles Young Dr S, Los Angeles, CA 90025. E-mail: hmiddlekauff@ 123456mednet.ucla.edu

                Author information
                https://orcid.org/0000-0002-9945-658X
                https://orcid.org/0000-0001-7848-8585
                Article
                JAH35527
                10.1161/JAHA.120.016983
                7726977
                32896211
                fe675b0e-e516-4b35-a844-eecb7ab32eca
                © 2020 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

                History
                : 10 April 2020
                : 13 August 2020
                Page count
                Figures: 6, Tables: 1, Pages: 12, Words: 19271
                Funding
                Funded by: Tobacco‐Related Disease Research Program
                Award ID: TRDRP 28IR‐0065
                Funded by: NIH National Center for Advancing Translational Science (NCATS) UCLA CTSI
                Award ID: L1TR001881
                Funded by: NIH , open-funder-registry 10.13039/100000002;
                Award ID: R01AG059501
                Award ID: R03AG059462
                Funded by: UCLA Center for AIDS Research
                Award ID: P30AI28697
                Categories
                Original Research
                Original Research
                Coronary Heart Disease
                Custom metadata
                2.0
                15 September 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.9.1 mode:remove_FC converted:30.09.2020

                Cardiovascular Medicine
                electronic cigarettes,monocytes,nicotine,reactive oxidative species,tobacco cigarettes,oxidant stress,inflammation

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