Abstract P2Y12 receptor (P2RY12), mainly expressed on platelets, is known for its central role in hemostasis. P2RY12 activation is also involved in cancer development through platelet adhesion to cancer cells supporting immune-evasion, promoting tumor angiogenesis and metastasis, among others. P2RY12 is known as an actionable target, and P2RY12 antagonists are in clinical use for cardiovascular diseases. However, very little data are available regarding the protein expression of P2RY12 in lung carcinomas. We performed immunohistochemical staining for P2RY12 in a cohort of non-small cell lung cancer (NSCLC) samples comprising 320 adenocarcinomas (LUAD) and 158 squamous cell carcinomas (LUSC). Results were evaluated using a dual approach combining microscopic assessment and digital image analysis (QuPath). Results were correlated with clinical-pathological data. We found significantly higher P2RY12 protein expression in LUSC compared to LUAD (p<0.001) via eyeballing (absent/low expression in 21.7% (34/158) and moderate/high expression in 78.3% (124/158) of LUSC cases versus absent/low expression in 98.4% (315/320) and moderate/high expression in 1.6% (5/320) of LUAD cases). Digital analysis yielded similar results. High P2RY12 expression was associated with a significantly better 5-year overall survival rate for the entire cohort (p=0.0048) as well as for the LUAD (p=0.015) and LUSC (p=0.05) subgroups. Furthermore, P2RY12 showed excellent discriminatory performance for classifying carcinomas as LUAD or LUSC, with an AUC of 0.916 in ROC-analysis. High P2RY12 expression is linked to a better prognosis and might serve as a promising novel prognostic biomarker for NSCLC. Its assessment could be implemented in future routine diagnostic workup. At the same time, the data suggest that P2RY12 could also serve as a diagnostic marker for LUSC.

Mit dem Rauchen aufzuhören ist schwierig. Noch schwieriger ist es, ohne Nikotinersatztherapie rauchfrei zu bleiben. In den letzten Jahren wurden verschiedene Alternativprodukte auf den Markt gebracht, darunter auch erhitzte Tabakerzeugnisse (HTP). In dieser Studie haben wir die akuten Auswirkungen des Konsums von IQOSTM und gloTM (2 HTP) auf die Funktion der kleinen Atemwege und die arterielle Steifigkeit in einem Head-to-Head-Design untersucht, im Vergleich mit brennenden Zigaretten, nikotinfreien E-Zigaretten und einer Scheinrauchergruppe. 17 gesunde Gelegenheitsraucher wurden in eine zentrale 5-armige Crossover-Studie eingeschlossen. Die Parameter der Funktion der kleinen Atemwege und der Hämodynamik wurden zu verschiedenen Zeitpunkten vor und nach dem Konsum mit dem Mobil-O-GraphTM (I.E.M., Stolberg, Deutschland) und dem TremoFlo® c-100 (THORASYS Thoracic Medical Systems Inc., Montreal, QC, Kanada) gemessen. Sowohl die Obstruktion der kleinen Atemwege als auch der Widerstand waren nach dem Konsum von Zigaretten und Tabakersatzprodukten signifikant erhöht. Alle nikotinhaltigen Produkte führten zu einem ähnlichen signifikanten Anstieg des Blutdrucks und der arteriellen Steifigkeit. Die hämodynamischen Parameter waren auch nach dem Konsum von nikotinfreien E-Zigaretten erhöht, aber der Anstieg war kürzer und schwächer als bei nikotinhaltigen Produkten. Wir kommen zu dem Schluss, dass HTP akute schädliche Auswirkungen auf die Funktion der kleinen Atemwege haben, die möglicherweise größer sind als die Auswirkungen brennender Zigaretten, obwohl die Gründe dafür noch nicht geklärt sind. Wie andere nikotinhaltige Produkte führen auch HTP zu einer akuten nikotinbedingten Erhöhung der arteriellen Steifigkeit und des kardiovaskulären Stresses, ähnlich wie bei brennenden Zigaretten, was diese Produkte mit einem erhöhten kardiovaskulären Risiko in Verbindung bringt.

Die Langzeitsauerstofftherapie beschreibt den Einsatz von verordnetem Sauerstoff über mindestens 16 Stunden pro Tag bei einer Grunderkrankung, die anhaltend bzw. chronisch zu einer hypoxämischen Insuffizienz führt. Dabei stehen vor allem die Reduktion der Letalität und die Verbesserung der Lebensqualität bei chronisch Erkrankten mit Sauerstoffmangel im Vordergrund des ärztlichen Handelns. Dieses erläutert der Beitrag anhand eines Flussdiagramms.

Background Smoking is a modifiable risk factor for SARS-CoV-2 infection. Evidence of smoking behavior during the pandemic is ambiguous. Most investigations report an increase in smoking. In this context, Google Trends data monitor real-time public information–seeking behavior and are therefore useful to characterize smoking-related interest over the trajectory of the pandemic. Objective This study aimed to use Google Trends data to evaluate the effect of the pandemic on public interest in smoking-related topics with a focus on lockdowns, vaccination campaigns, and incidence. Methods The weekly relative search volume was retrieved from Google Trends for England, Germany, Italy, and Spain from December 31, 2017, to April 18, 2021. Data were collected for keywords concerning consumption, cessation, and treatment. The relative search volume before and during the pandemic was compared, and general trends were evaluated using the Wilcoxon rank-sum test. Short-term changes and hereby temporal clusters linked to lockdowns or vaccination campaigns were addressed by the flexible spatial scan statistics proposed by Takahashi and colleagues. Subsequently, the numbers of clusters after the onset of the pandemic were compared by chi-square test. Results Country-wise minor differences were observed while 3 overarching trends prevailed. First, regarding cessation, the statistical comparison revealed a significant decline in interest for 58% (7/12) of related keywords, and fewer clusters were present during the pandemic. Second, concerning consumption, significantly reduced relative search volume was observed for 58% (7/12) of keywords, while treatment-related keywords exhibited heterogeneous trends. Third, substantial clusters of increased interest were sparsely linked to lockdowns, vaccination campaigns, or incidence. Conclusions This study reports a substantial decline in overall relative search volume and clusters for cessation interest. These results underline the importance of intensifying cessation aid during times of crisis. Lockdowns, vaccination, and incidence had less impact on information-seeking behavior. Other public measures that positively affect smoking behavior remain to be determined.

INTRODUCTION:Lung cancer is a major cause of cancer-related death worldwide and effective therapies, besides surgery, are available only for a small proportion of patients. Since cellular respiration is known to be broadly altered in malignant tumors, the cellular processes of respiration can be a potential therapeutic target. One important element of cellular respiration is creatine and its transport by the creatine transporter SLC6A8. Here we describe the expression of SLC6A8 at the RNA and protein level, epigenetic modifications as well as survival analysis in NSCLC tissues and matched controls. MATERIALS AND METHODS:We analyzed epigenetic modifications of the SLC68A gene in 32 patients, of which 18 were additionally analyzed by transcriptome analysis. The expression of SLC6A8 at the protein level was assessed by immunohistochemistry using an independent cohort and correlated with clinicopathological data including survival. Kaplan-Meier analysis was performed to analyze the possible effects of the transcriptional levels of SLC6A8 in another separate cohort (n=1925). RESULTS:SLC6A8 loci are epigenetically modified in NSCLC compared with tumor-free controls. SLC6A8 is upregulated in NSCLC at the RNA and protein level. High mRNA expression of SLC6A8 was associated with an overall poor prognosis in lung adenocarcinoma patients and displayed the strongest adverse prognostic effect in male smokers with adenocarcinomas. Results of transcriptome analysis were partially confirmed at the protein level. CONCLUSIONS:Our results suggest an important role of creatine and its transport via SLC6A8 in NSCLC.

BACKGROUND/AIM:Tobacco is a carcinogen that is closely associated with the occurrence of lung cancer and head and neck squamous cell carcinoma (HNSCC). The consumption of tobacco is also leading to alterations in different immune cell subtypes. However, the impact of different conventional and alternative smoking sources on human monocytes remains elusive.MATERIALS AND METHODS:In this study, we investigated the influence of aqueous extracts of different sources of smoking (cigarettes; heated tobacco product IQOS; e-cigarettes with and without nicotine; nicotine pouches) on different monocytic adhesion molecules, chemokine receptors and checkpoint molecule PD-L1 by flow cytometry. Cytokine expression patterns were evaluated using human cytokine arrays and the human monocyte leukemia cell line THP-1 as a model.RESULTS:Data revealed differential effects of the analyzed conventional and alternative smoking devices on monocyte adhesion molecules and cytokine secretion. The examined smoking devices can be assigned to two differential monocyte activation patterns. Monocytes stimulated with aqueous extracts of cigarettes, e-cigarette without nicotine, and heat not burn product IQOS revealed distinct alterations of surface markers and cytokines compared to the monocyte activation pattern in response to aqueous extracts of nicotine, nicotine pouches, and e-cigarette with nicotine.CONCLUSION:Our data indicate differential immunological consequences of different conventional and alternative smoking sources with and without nicotine. Further comprehensive analysis as well as in vivo investigations on peripheral blood monocyte subsets from smoking individuals using different smoking sources are required to better understand the impact on monocyte characteristics, especially with regard to the development of cancer.

Introduction E-cigarettes continue to be controversial in science and especially for smoking cessation. A new trend has spread among young people and adolescents – disposables, in which the nicotine-containing liquid is in a cartridge as a disposable product and advertises with colorful and fruity aromas. We investigated the acute effects of two different ELFBAR disposables head-to-head to combustible cigarette and a e-Cigarettes (MyBlu) with nicotine (18 mg/ml) on vascular function.

In addition to the market launch of heated tobacco products (HTPs) and the JUUL as well as the EVALI, they caused a widespread discussion on the risk reduction compared to a combustible cigarette. Furthermore, first data showed harmful effects on the cardiovascular system. We, therefore, conducted investigations including a control group with a nicotine-free liquid. Forty active smokers were studied in two different approaches during and after consuming an HTP, a cigarette, a JUUL, or a typical electronic cigarette with or without nicotine in a partly double-blinded randomised, cross-over trial. Inflammation, endothelial dysfunction, and blood samples (full blood count, ELISA, multiplex immunoassay) were analysed, and arterial stiffness was measured. In addition to the cigarette, an increase in the white blood cell count but also in proinflammatory cytokines was shown for the various nicotine delivery systems. These correlated with the parameters of arterial vascular stiffness as a clinical parameter of endothelial dysfunction. It can be shown that even a single consumption of the different nicotine delivery system or cigarette leads to a significant inflammatory reaction followed by endothelial dysfunction and increased arterial stiffness causing cardiovascular disease. Inflammation, endothelial dysfunction, and arterial stiffness should be addressed in long-term observational studies.

Cardiovascular health at a young age has implications for preventing cardiovascular disease, and it is associated with improved physical and cognitive performance during the aging process. Sports are well known to prevent cardiovascular disease; however, school-based interventions have mostly been neglected. This cross-sectional study aimed to compare groups of high school students, stratified by the amount of physical activity in their high school curriculum and downtime. Comparisons concerning physical and cognitive performance and arterial stiffness were made. A total of 63 senior-year students were investigated. Arterial stiffness was assessed using the oscillometric technique with ArteriographTM detection. Three-kilometer and pendulum runs were conducted as typical training loads. Cognitive performance was evaluated via the visual and verbal memory and number connection tests. Regarding cognitive skills, extracurricular physical activity improved the number connection test in male participants (p = 0.004). For physical performance, female students with a sports-focused curriculum were faster in the 3 km run (p < 0.001). Concerning arterial stiffness, the measurements yielded a lower mean arterial pressure (p = 0.015) and aortic pulse wave velocity (p = 0.04) in male students with a sports-focused curriculum. In summary, extracurricular physical activity and enrollment in a sports-focused curriculum may be associated with lower cardiovascular risk due to lower arterial stiffness and better physical and cognitive abilities.

Rationale: Telemonitoring of lung function and symptoms might help to identify exacerbations to prevent hospitalizations. We investigated the acceptance and usability of telemonitoring in COPD patients. Methods: In this prospective multicentre study we used home-based spirometry, accelerometry, a smartphone-based disease-control interface (DCI) for symptoms: CAT questionnaire, need for rescue inhaler. We evaluated individual changes of FEV1 beyond day-to-day variability, steps, change in CAT score, rescue inhaler use for 12 months follow-up after exacerbation Results: 43 patients (n=43; mean age, 65±7 years; mean FEV1%, 42± 14) were enrolled. Adherence to home-based spirometry, DCI and accelerometry was 247, 252 and 58 days, respectively. We found a good agreement between in-clinic and home-based spirometry (Bland–Altman: mean difference 111 ml; 95% limits of agreement: −506 to 782 ml). In patients with exacerbation during follow-up (n=17), home-based spirometry revealed a mean decline of 152 ml in FEV1 vs. stable FEV1 (+22 ml) in those without exacerbation, (p=0.056). Changes in home-based FEV1 showed a stronger correlation with time to first exacerbation and number of exacerbations (correlation coefficients 0.38 and -0.25) than in-clinic based-FEV1. There was a good correlation between both components of DCI (R=0.36, p=0.044). Clinically relevant changes in FEV1 and DCI could be observed 5 days right before and during exacerbations. Conclusion: In adherent COPD patients home-based telemonitoring provides plausible and reliable changes of symptoms and lung function that might allow an early intervention.

Dysregulated hyperinflammatory response is key in the pathogenesis in patients with severe COVID-19 leading to acute respiratory distress syndrome and multiorgan failure. Whilst immunosuppression has been proven to be effective, potential biological targets and optimal timing of treatment are still conflicting. We sought to evaluate efficacy and safety of the Janus Kinase 1/2 inhibitor ruxolitinib, employing the previously developed COVID-19 Inflammation Score (CIS) in a prospective multicenter open label phase II trial (NCT04338958). Primary objective was reversal of hyperinflammation (CIS reduction of ≥25% at day 7 in ≥20% of patients). In 184 patients with a CIS of ≥10 (median 12) ruxolitinib was commenced at an initial dose of 10 mg twice daily and applied over a median of 14 days (range, 2–31). On day 7, median CIS declined to 6 (range, 1–13); 71% of patients (CI 64–77%) achieved a ≥25% CIS reduction accompanied by a reduction of markers of inflammation. Median cumulative dose was 272.5 mg/d. Treatment was well tolerated without any grade 3–5 adverse events related to ruxolitinib. Forty-four patients (23.9%) died, all without reported association to study drug. In conclusion, ruxolitinib proved to be safe and effective in a cohort of COVID-19 patients with defined hyperinflammation.

Book Citations: Authors, Title, HemaSphere, 2023;7(S3):pages. The individual abstract DOIs can be found at https://journals.lww.com/hemasphere/pages/default.aspx. Disclaimer: Articles published in the journal HemaSphere exclusively reflect the opinions of the authors. The authors are responsible for all content in their abstracts including accuracy of the facts, statements, citing resources, etc. 2996 (especially IL-6) was demonstrated. Toxicity comprised elevated liver enzymes (8.5 %) and bleeding complications (3.3%), but no severe adverse events in relationship to Rux were reported. Forty-four pts (23.9 %) died, in most cases due to progression of Covid19.

Background: The widespread use of the JUUL™ device ignited a discussion about the effects these products have on harm reduction. Therefore, we conducted a study directly comparing the JUUL™ device with a cigarette, a heated tobacco product, and a nicotine-free e-cigarette to examine the acute effects on arterial stiffness. Methods: This crossover-designed study examines 20 occasional smokers (age 25.2 ± 2.5 years). Study participants used each of the four smoking devices for a duration of 5 min following a protocol. Peripheral blood pressure and parameters of arterial stiffness and endothelial vasodilator function such as the reactive hyperemia index and the augmentation index were measured using the EndoPAT™2000 before and after. Results: In addition to significant peripheral hemodynamic changes after 5 and 10 min (p < 0.05), the reactive hyperemia index showed a significant decrease for all devices 15 min after consumption and remained significantly decreased after 60 min (p < 0.01). The augmentation index adjusted for a heart rate of 75 bpm increased significantly for all devices 15 and 60 min after consumption (p < 0.01). Conclusions: In conclusion, the increases in blood pressure and arterial stiffness are similar after smoking, JUUL™ing, heating, and vaping. These changes may be associated with an increase in cardiovascular risks; however, an evaluation of the long-term effects of JUUL™ing, vaping and heating is needed.

(1) Background: watching sporting events may trigger cardiovascular events by elevating emotional stress levels. The underlying reasons and specific populations at risk are not well defined. (2) Methods: we conducted a multicenter prospective trial at three German sites during the UEFA Soccer EC 2012 and 2021 comprising 52 healthy participants (noCVD) and 18 patients hospitalized with cardiovascular disease (CVD). Subjects were studied during matches of the German national team (GP) as well as corresponding matches without German participation (noGP). Peripheral and central blood pressure (BP) and parameters of arterial stiffness were measured (Mobil-O-Graph™, I.E.M., Stolberg, Germany) before, during, and after the matches. (3) Results: in terms of CVD, peripheral as well as central BP and heart rate increased significantly during GP as well as noGP matches and remained elevated beyond the end of the matches. Likewise, arterial stiffness parameters and vascular resistance were higher during the matches and remained elevated after the matches. No consistent significant differences were found between GP and noGP matches. (4) Conclusions: this is the first study on real-life changes in hemodynamics during sport-associated emotional stress, with comparison between noCVD and CVD. We found that alterations were profound in CVD and remained elevated even after the matches.

To control the ongoing global pandemic due to SARS-CoV-2, we need to influence people's behavior. To do so, we require information on people's knowledge and perception of the disease and their opinions about the importance of containment measures. Therefore, in August 2020, we conducted an anonymous cross-sectional online survey on these topics in 913 participants in Germany. Participants completed a questionnaire on various synonyms and symptoms of corona virus and specified the importance they attributed to individual and regulatory measures. The virus was linked more closely with most synonyms and the discovery in China than with the places of the first larger European outbreaks. General (cold-like) symptoms, such as "cough" and "fever," were more widely known than COVID-19-specific ones, e.g., "loss of taste and smell." The widely promoted individual measures "distancing," "hygiene," and "(facial) mask wearing" were rated as highly important, as were the corresponding official measures, e.g., the "distancing rule" and "mask mandate." However, the "corona warning app" and a "vaccine mandate" were rated as less important. A subgroup analysis showed broad agreement between the subgroups on nearly all issues. In conclusion, the survey provided information about the German population's perception and knowledge of the coronavirus five months into the pandemic; however, participants were younger and more educated than a representative sample. To learn from the beginning and still ongoing pandemic and develop concepts for the future, we need more conclusive studies, especially on the acceptance of further specified lockdowns, the population's willingness to be vaccinated, and the influence of misinformation on public opinion.

Introduction The rapid growth in the e-cigarette market after the launch of JUUL e-cigarettes led to much discussion on the potential benefits and risks of pods, JUUL devices, and conventional e-cigarettes compared with combustible cigarettes. Independent data are required to assess the effects of these products on cardiovascular surrogate parameters and cardiovascular risk. Methods We conducted a single-center three-arm study comparing combustible cigarettes with JUUL e-cigarettes with the old and new technology. We recruited 32 participants who were active smokers (n=15) or vapers (n=17) and performed a total of 39 measurements before and 5, 15, and 30 minutes, after participants smoked a combustible cigarette or vaped a JUUL e-cigarette with the new or old technology. Measurements included peripheral and central blood pressures and parameters of arterial stiffness, including pulse wave velocity and augmentation index. Results Peripheral systolic blood pressure, central blood pressure, and peripheral pulse rate increased significantly in all three groups (each p<0.05). Heart rate (HR) changes lasted significantly longer than blood pressure changes. The augmentation index and pulse wave velocity increased in all three groups, and a multivariate analysis of variance showed that the increases were independent of systolic blood pressure, sex, age, device, and HR. Conclusions Changes in blood pressure and arterial stiffness are similar after cigarette smoking and JUUL use. These changes may be associated with an increased cardiovascular risk compared with no product use. However, a longterm follow-up evaluation of JUUL use and a head-to-head comparison with conventional e-cigarettes are still needed.

Three-dimensional tracking of cells is one of the most powerful methods to investigate multicellular phenomena, such as ontogenesis, tumor formation or wound healing. However, 3D tracking in a biological environment usually requires fluorescent labeling of the cells and elaborate equipment, such as automated light sheet or confocal microscopy. Here we present a simple method for 3D tracking large numbers of unlabeled cells in a collagen matrix. Using a small lensless imaging setup, consisting of an LED and a photo sensor only, we were able to simultaneously track ~3000 human neutrophil granulocytes in a collagen droplet within an unusually large field of view (>50 mm2) at a time resolution of 4 seconds and a spatial resolution of ~1.5 μm in xy- and ~30 μm in z-direction. The setup, which is small enough to fit into any conventional incubator, was used to investigate chemotaxis towards interleukin-8 (IL-8 or CXCL8) and N-formylmethionyl-leucyl-phenylalanine (fMLP). The influence of varying stiffness and pore size of the embedding collagen matrix could also be quantified. Furthermore, we demonstrate our setup to be capable of telling apart healthy neutrophils from those where a condition of inflammation was (I) induced by exposure to lipopolysaccharide (LPS) and (II) caused by a pre-existing asthma condition. Over the course of our experiments we have tracked more than 420.000 cells. The large cell numbers increase statistical relevance to not only quantify cellular behavior in research, but to make it suitable for future diagnostic applications, too.

Smoking is considered a major preventable cause of cardiovascular and lung diseases, as well as cancer. During the COVID-19 pandemic, there was extensive discussion about the influence of nicotine use; ultimately, smoking was considered a major risk factor for poor disease progression. Therefore, in April 2021, we conducted an anonymous cross-sectional online survey on smoking and vaping behavior, as well as smoking cessation, in four different countries in Europe (the United Kingdom, Germany, Spain, and Italy). A total of 3605 participants completed a questionnaire on their smoking and vaping behaviors and smoking cessation because of and during the COVID-19 pandemic. Fear of COVID-19 infection, a high percentage of quarantine stays (44.9% Italy and 52.1% Spain), and high infection (75.5% Italy and 52.4% Spain) and death (42% Italy) rates in respondents' personal circles were observed mostly in the surveyed populations of Italy and Spain. Smoking cessation attempts and success were mainly seen in the Italian population and were linked to psychological distress, while the same effects were shown for vaping in Spain. In summary, health anxiety was detected in all cohorts. Despite these findings, smoking as a risk factor for severe outcomes of COVID-19 infection did not lead to a higher rate of smoking cessation attempts.