Blood Of Smokers Research Articles

Leukocyte- and Platelet-Rich Fibrin (L-PRF) Obtained from Smokers and Nonsmokers Shows a Similar Uniaxial Tensile Response In Vitro.

We evaluated and compared the biomechanical properties of Leukocyte-and Platelet Rich Fibrin L-PRF clots and membranes derived from smoker and nonsmoker donors. Twenty venous-blood donors (aged 18 to 50 years) were included after signing informed consent forms. L-PRF clots were analyzed and then compressed to obtain L-PRF membranes. L-PRF clot and membrane samples were tested in quasi-static uniaxial tension and the stress-stretch response was registered and characterized. Furthermore, scanning electron microscope representative images were taken to see the fibrin structure from both groups. The analysis of stress-stretch curves allowed us to evaluate the statistical significance in differences between smoker and nonsmoker groups. L-PRF membranes showed a stiffer response and higher tensile strength when compared to L-PRF clots. However, no statistically significant differences were found between samples from smokers and nonsmokers. With the limitations of our in vitro study, we can suggest that the tensile properties of L-PRF clots and membranes from the blood of smokers and nonsmokers are similar. More studies are necessary to fully characterize the effect of smoking on the biomechanical behavior of this platelet concentrate, to further encourage its use as an alternative to promote wound healing in smokers.

Circular RNA 0000157 depletion protects human bronchial epithelioid cells from cigarette smoke extract-induced human bronchial epithelioid cell injury through the microRNA-149-5p/bromodomain containing 4 pathway.

Circular RNA (circRNA) has been reported to regulate respiratory diseases. In the study, we aimed to elucidate the role of circ_0000157 in smoke-related chronic obstructive pulmonary disease (COPD) and the inner mechanism. COPD-like cell injury was induced by treating human bronchial epithelioid cells (16HBE) with cigarette smoke extract (CSE). The expression of circ_0000157, miR-149-5p, bromodomain containing 4 (BRD4), BCL2-associated x protein (Bax) and B-cell lymphoma-2 (Bcl-2) was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) or Western blotting. Enzyme-linked immunosorbent assay was performed to detect interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels. Malondialdehyde (MDA) production was detected by a lipid peroxidation MDA assay kit. Superoxide dismutase (SOD) activity was analyzed by a SOD activity assay kit. Circ_0000157 and BRD4 expression were upregulated, while miR-149-5p expression was downregulated in the blood of smokers with COPD and CSE-induced 16HBE cells compared with control groups. CSE treatment inhibited 16HBE cell proliferation and induced cell apoptosis, inflammation, and oxidative stress; however, these effects were remitted when circ_0000157 expression was decreased. In addition, circ_0000157 acted as a miR-149-5p sponge and regulated CSE-caused 16HBE cell damage by targeting miR-149-5p. The overexpression of BRD4, a target gene of miR-149-5p, attenuated the inhibitory effects of miR-149-5p introduction on CSE-induced cell damage. Further, circ_0000157 modulated BRD4 expression by associating with miR-149-5p in CSE-treated 16HBE cells. Circ_0000157 knockdown ameliorated CSE-caused 16HBE cell damage by targeting the miR-149-5p/BRD4 pathway, providing a potential therapeutic strategy for clinic intervention in COPD.

Do Circulating Extracellular Vesicles Strictly Reflect Bronchoalveolar Lavage Extracellular Vesicles in COPD?

Cell-derived extracellular vesicles (EVs) found in the circulation and body fluids contain biomolecules that could be used as biomarkers for lung and other diseases. EVs from bronchoalveolar lavage (BAL) might be more informative of lung abnormalities than EVs from blood, where information might be diluted. To compare EVs' characteristics in BAL and blood in smokers with and without COPD. Same-day BAL and blood samples were obtained in 9 nonsmokers (NS), 11 smokers w/o COPD (S), and 9 with COPD (SCOPD) (FEV1: 59 ± 3% pred). After differential centrifugation, EVs (200-500 nm diameter) were identified by flow cytometry and labeled with cell-type specific antigens: CD14 for macrophage-derived EVs, CD326 for epithelial-derived EVs, CD146 for endothelial-derived EVs, and CD62E for activated-endothelial-derived EVs. In BAL, CD14-EVs were increased in S compared to NS [384 (56-567) vs. 172 (115-282) events/μL; p = 0.007] and further increased in SCOPD [619 (224-888)] compared to both S (p = 0.04) and NS (p < 0.001). CD326-EVs were increased in S [760 (48-2856) events/μL, p < 0.001] and in SCOPD [1055 (194-11,491), p < 0.001] when compared to NS [15 (0-68)]. CD146-EVs and CD62E-EVs were similar in the three groups. In BAL, significant differences in macrophage and epithelial-derived EVs can be clearly detected between NS, S and SCOPD, while these differences were not found in plasma. This suggests that BAL is a better medium than blood to study EVs in lung diseases.

EVALUATION OF ANTIOXIDANTS ENZYMES AND AUTOPHAGY GENES IN THE BLOOD OF IRAQI SMOKER VOLUNTEERS.

The aim: In this study, we investigated the expression level of LC3A, LC3B, Beclin1, and ATG5 in the blood of smokers as well as non-smokers volunteers. On the other hand, total antioxidant status (TAS), catalase (CAT), and superoxide dismutase (SOD) in the same individuals were measured. Material and methods: About 5 ml of blood was collected from 55 smokers (workers in a coffee shop) and 24 non-smokers volunteers. The blood was divided into two parts: 2.5 ml collected in a K2-EDTA tube that was used for RNA extraction to estimate the autophagic genes including LC3A, LC3B, Beclin1, and ATG5 by using of qRT-PCR and for hematological studies. The other part - was centrifuged to obtain the serum samples to measure TAS, CAT, and SOD by spectrophotometrical method. Results: The fold expression of autophagy genes was significantly increased in the blood of smokers as compared to non-smokers' individuals at (p ≤ 0.05). On the other hand, TAS, CAT, and SOD were significantly increased in smokers as compared with non-smokers individuals at (p ≤ 0.05). The hematological study revealed increased hemoglobin concentration and hematocrit in the smoker's blood as compared to nonsmokers' individuals. No significant variation was found in other hematological parameters in the smoker's individual as compared to non-smokers at (p ≤ 0.05). Conclusions: Our results indicate that smoke exposure may induce autophagy genes through increased antioxidant status and antioxidant enzymes, and the better way to be healthy is to quitting smoking.

Malondialdehyde-Induced Post-translational Modifications in Hemoglobin of Smokers by NanoLC-NSI/MS/MS Analysis.

Malondialdehyde (MDA) is the most abundant α,β-unsaturated aldehyde generated from endogenous peroxidation of polyunsaturated fatty acids and is present in cigarette smoke. Post-translational modifications of blood hemoglobin can serve as biomarkers for exposure to chemicals. In this study, two types of MDA-induced modifications, the N-propenal and the dihydropyridine (DHP), were identified at multiple sites in human hemoglobin digest by the high-resolution mass spectrometry. The N-propenal and the DHP types of modification led to the increase of 54.0106 and 134.0368 amu, respectively, at the N-terminal and lysine residues. Among the 21 MDA-modified peptides, 14 with dose-response to MDA concentrations were simultaneously quantified in study subjects by the nanoflow liquid chromatography nanoelectrospray ionization tandem mass spectrometry under selected reaction monitoring (nanoLC-NSI-MS/MS-SRM) without prior enrichment. The results showed that the modifications of the N-propenal-type at α-Lys-11, α-Lys-16, α-Lys-61, β-Lys-8, and β-Lys-17, as well as the DHP-type at the α-N-terminal valine, are significantly higher in hemoglobin isolated from the blood of smokers than in nonsmoking individuals. This is the first report to identify and quantify multiple sites of MDA-induced modifications in human hemoglobin from peripheral blood. Our results suggest that the MDA-derived modifications on hemoglobin might represent valuable biomarkers for MDA-induced protein damage.

The Effect of Living Place and Smoking on Blood Level of Lead, Cadmium and Mercury: As a Model of Pollutants in Human Blood

It is known that heavy metals (as lead, cadmium and mercury) are naturally occurring elements that have a high atomic weight and a density 5 times greater than that of water. Their different industrial, domestic, agricultural, medical and technological applications may led to distribution widely in the environment, raising their harmful effects on human health and the environment. Their effects and degree of toxicity depend on several factors, including the level of these metals in the blood and the age, gender, genetics, smoking, living place, and nutritional status of exposed individuals. Accordingly and in fact is needed to estimate heavy metals levels in the blood of living people in different areas in Iraq from time to time. The present study was designed to search the relation between heavy metal (lead, cadmium, and mercury) levels in the blood of people and smoking, and living place (rural, and city center), gender, as factors that may aggravate heavy metal toxicity in Diyala Province. Total 72 patients were considered in this study. Group (A) of 21 males (A1, A2, A3, A4, A5, A6, A7) each sub group compose of three males, A1, A2, A3, from rural areas, while A4, A5, A6, A7,were from the center of the city, and group (B) of 51 females ( B1. B2. B3, B4) each sub group of 12, 13, 8, 18 in number respectively according to availability, at age (≤ 29), and at age (≥30). The results showed that in comparison 1 the level of lead and cadmium in the blood of individuals living in the center of the city was significantly higher in comparison to those living in the village in both mail and females at age (≤ 29), While in comparison 2, it was noted that the level of lead and cadmium together in the villagers was more than its level in the blood of city resident individuals, but statistically was not significant. The city’s residents than those in rural areas had more levels of mercury in their blood in all comparisons but statistically was not significant between the studied groups. The smoking factor had an effect on the levels of heavy metals in the blood, as the level of lead in smokers was higher compared to its level in non-smokers (control group) in comparisons 3 and 1, while the opposite was in comparison 2, the level of lead increased in non-smokers but statistically was not significant. The level of cadmium in the blood of smokers was higher compared to its level in non-smokers in all comparisons, as well as with mercury, its level increased in smokers in comparisons 3 and 1, except for comparison 2, where its level increased in non-smokers. The concentration of lead and mercury did not exceed the internationally permissible limits in individuals’ blood, while the cadmium level exceeded the internationally permissible limits.

N-OH-AABP Modifications in Human DNA May Lead to Auto-Antibodies in Bladder Cancer Subjects.

4-Aminobiphenyl (4-ABP) and other related arylamines have emerged to be responsible for human urinary bladder tumors and cancers. Hemoglobin-ABP adducts have been recognized in the blood of smokers, and it builds up in the circulatory system over the period of years that might lead to a bladder tumor. N-hydroxy-Acetyl 4-Aminobiphenyl (N-OH-AABP) is one of the reactive forms of 4-ABP which has a potential to initiate tumor growth and causes cancer rapidly. In the present study, commercially available human DNA was modified by N-OH-AABP, and its modifications were analyzed biophysically from fluorescence spectroscopy and thermal denaturation studies. Further, Sera and IgG from bladder cancer patients’ blood were assessed for affinity to native and N-OH-AABP modified human DNA using ELISA. The study showed N-OH-AABP caused damage in the structure of the DNA macromolecule and the perturbations resulting from damage leads to change in the Tm of the DNA molecule. Bladder cancer auto-antibodies, particularly in smoker group, showed preferential binding to N-OH-AABP modified human DNA. This study shows that N-OH-AABP modified DNA could be an antigenic stimulus for the generation of autoantibodies in the sera of bladder cancer patients.

Epigenetic Age Acceleration Is Not Associated with Age-Related Macular Degeneration.

DNA methylation age (DNAm age) estimation is a powerful biomarker of human ageing. To date, epigenetic clocks have not been evaluated in age-related macular degeneration (AMD). Here, we perform genome-wide DNA methylation analyses in blood of AMD patients with a documented smoking history (14 AMD, 16 Normal), identifying loci of differential methylation (DML) with a relaxed p-value criterion (p ≤ 10−4). We conduct DNAm age analyses using the Horvath-multi tissue, Hannum and Skin & Blood epigenetic clocks in both blood and retinal pigment epithelium (RPE). We perform Ingenuity Pathway Analysis Causal Network Analysis (IPA CNA) on the topmost significantly differentially methylated CpG probes in blood and RPE. Results show poor performance of epigenetic clocks in RPE. Epigenetic age acceleration (EAA) was not observed in AMD. However, we observe positive EAA in blood of smokers, and in smokers with AMD. DML analysis revealed hypomethylation at cg04953735 within RPTOR (p = 6.51 × 10−5; Δβ = −11.95%). IPA CNA in the RPE also identified RPTOR as the putative master regulator, predicted to be inhibited in AMD. In conclusion, this is the first study evaluating an association of epigenetic ageing in AMD. We posit a role for RPTOR as a common master regulator of methylation changes in the RPE in AMD.

Determination of carboxyhaemoglobin and methaemoglobin levels in donated blood

To determine the percentages of carboxyhaemoglobin (COHb) and methaemoglobin (MeHb) in donor blood and to compare these levels between smokers and nonsmokers at different time points during blood storage. Blood donors were recruited from Haematology Service, University Hospital Alzira Velano, Alfenas-MG. The blood was kept in collecting ducts (noodles) containing citrate, phosphate and dextrose (CPD) and stored at 4°C throughout the storage period. Since the noodles kept the characteristics of the bags, COHb and MeHb levels were analysed on the day of donation and after 20 days of storage. Levels of COHb and MeHb were determined using spectrophotometric methods. Non-parametric Friedman and Mann-Whitney tests were employed to compare COHb and MeHb levels before and after the storage and groups of smokers and nonsmokers, respectively. Levels of COHb and MeHb in the blood collected from smokers and nonsmokers were statistically different (p&lt; 0.05; Mann- Whitney test) when the samples were analyzed before the storage. In blood of smokers, COHb levels were no different over a 20-day storage period (p= 0.7009; Friedman test). On the other hand, MeHb levels were significant different over a 20-day storage period (p&lt; 0.05). The results suggest the need to regularly assess COHb and MeHb levels in donor blood stored in blood banks.

Activity of glutathione S-transferase and its π isoenzyme in the context of single nucleotide polymorphism in the GSTP1 gene (rs1695) and tobacco smoke exposure in the patients with acute pancreatitis and healthy subjects

Oxidative stress associated with the course of acute pancreatitis (AP) can cause changes in the involvement of antioxidants, which can result in the increased production of free radicals with pro-inflammatory potential. Through its noncatalytic activity, the glutathione S-transferase and its π isoenzyme (GST-π), apart from cellular xenobiotics detoxification, are involved in the regulation of cellular signalling, metabolism and apoptosis. This study aimed to evaluate the impact of SNP rs1695 in the GSTP1 gene on GST and GST-π activity in healthy subjects and patients with acute pancreatitis (AP). The concentration of glutathione (GSH) as an important component of the antioxidant system, necessary for environmental xenobiotics detoxification by GST, and malonyldialdehyde (MDA) as a marker of oxidative stress induced by inflammation were also assessed. SNP was examined in 39 AP patients and 51 healthy subjects using PCR-RFLP methods. GST activity (in plasma and erythrocyte lysate) and GST-π activity (in erythrocyte lysate) were measured using the spectrophotometric method with 1-chloro-2,4-dinitrobenzene and ethacrynic acid as substrate, respectively. Blood GSH concentration was measured using the Patterson method. Concentrations of high-sensitive C-reactive protein (hs-CRP) and MDA were measured using commercial tests. In the blood of non-smoking AP patients with GG genotypes for SNP rs1695 in the GSTP1 gene, the lowest GST-π activity was shown. It was accompanied by the lowest hsCRP concentration in this group. In the blood of smoking healthy subjects with AG genotype, a decrease in GST-π activity was noted compared to non-smokers from this group. However, in the blood of smokers with AP, a gradually decreasing GST-π activity was noted in individuals with AA genotype, which was associated with the increasing MDA concentration. It confirms the role of GST-π in the neutralization of oxidative stress induced by the exposure to smoke xenobiotics.

Estimated of U, Rn and Po Concentrations in Smokers Blood Samples Collected from Babylon, Iraq

Because of the greats pressures on humans, there are resorted to smoking without knowing its effects on health. Such symptoms include adult lung cancer, cardiovascular, respiratory infections, kidney failure, congenital abnormalities and chronic bronchitis. Uranium and radon estimation has carried out in 100 smoker and Non-smoker blood samples from Babylon, Iraq, using solid state nuclear track detector (CR-39). The results have been shown the total uranium concentrations in blood smokers and non-smokers samples to be 1.02 ± 0.26 ppb and 0.86 ± 0.17 ppb respectively. On the other side, the total concentrations of radon in blood smokers and non-smokers samples 4.98 ± 0.79 Bqm−3 and 3.59 ± 0.28 Bqm−3 respectively. The total concentrations of uranium and radon in blood samples of smokers are greater than non- smokers blood samples. While, the total polonium (POW) and (POS) for smoker and non- smoker samples to be 0.527 KBqm−3 - 0.127 KBqm−3 and from 0.378 KBqm−3 to 0.091 KBqm–3 respectively. These studies conclude the smoking causes an increase in the concentration of uranium and radon and polonium in the blood of smokers.

Study of Toxic and Non-Toxic Metals in the Blood of Smokers and Non-Smokers by Atomic Absorption Spectroscopy

Cigarette smoking is considered as addictive and destructive behaviour. It creates health problems and variation in different parameters of blood. Cigarette smoking also causes environmental pollution by releasing toxic air pollutants into the atmosphere. Analysis of Lead (Pb), Cadmium (Cd) and Iron (Fe) in blood of smokers and non-smokers was done by Atomic Absorption Spectroscopy and there was also comparison of effect of cigarette smoking on haemoglobin (Hb) between smokers and non-smokers before and after medication. Samples of smokers and non-smokers were collected from different areas of Lahore. After experimentation, statistics applied for conclusion. Samples of smokers and non-smokers were collected to compare the effect of smoking on Hb before and after medication. Smokers and non-smokers were medicated by medicine Surbex Z for one week. The Hb level of smokers and nonsmokers was noted before and after medication. Time duration has been taken into account for effective medication.

Analysis of the Status of the Cutaneous Endogenous and Exogenous Antioxidative System of Smokers and the Short-Term Effect of Defined Smoking Thereon.

The daily consumption of tobacco products leads to a boost in free radical production in tissues, promoting the risk for malignancies, metabolic alterations and chronic-inflammatory diseases. This study aimed to broaden the knowledge of the status of the antioxidative (AO) system in the skin, compared to the blood, of healthy appearing smokers. Both, the basic status compared to non-smokers and the short-term impact of controlled cigarette consumption in smokers were analyzed. Our study showed that the basic level of the AO system of smokers significantly differed from that of non-smokers. As determined by resonant Raman spectroscopy (RRS), the levels of exogenous AOs were decreased in both, the skin, in vivo (β-carotene and lycopene), and blood plasma (β-carotene only). In contrast, the levels of glutathione (GSH), the prototypical endogenous AO, which were analyzed by fluorimetric assays in cutaneous tape strips and blood plasma, were increased in the skin, although unchanged in the blood of smokers. Elevated cutaneous GSH levels were reflected by an elevated overall radical scavenging activity in the skin, as quantified by non-invasive electron paramagnetic resonance (EPR) spectroscopy. Analysis of the expression of selected stress-associated genes in blood immune cells by quantitative RT-PCR in subgroups of non-smokers and smokers additionally demonstrated the downregulation of AKR1C2 in smokers, and its negative correlation with blood plasma levels of the protective immune mediator interleukin-22, assessed by the ELISA technique. Controlled cigarette consumption did not alter exogenous or endogenous AOs in the skin of smokers, but decreased lycopene levels in blood plasma. Moreover, there was a decline in blood IL-22 levels, while no relevant response of blood cell gene expressions was found after the considered short time. Our data therefore demonstrate a strengthened endogenous AO status in the skin of smokers, which may indicate a long-term adaptation to chronic oxidative stress in this specific organ. While this effect was not clearly visible in the blood, this compartment seems to be useful as an immediate indicator of the body’s AO consumption. Moreover, decreased levels of AKR1C2, which we show for the first time to be expressed in immune cells, may be a candidate marker for long-term smoking. In addition, this study demonstrates that the rate constant of a spin probe decline determined by EPR spectroscopy mainly represents the endogenous AO status of a tissue.

The influence of smoking habits on cadmium and lead blood levels in the Serbian adult people.

According to the World Health Organization, in 2015, the Serbian population ranked among the highest ones in Europe in terms of smoking habit: 44.3% males and 36.2% females aged 18-64 smoked tobacco. In the last 7 years, 25% of total mortality in men and 9% in women from Serbia were associated with smoking. Tobacco smoking is one of the most important sources of exposure to many toxic substances in general population. Our study confirmed higher blood levels of two toxic metals, cadmium and lead, in the blood of smokers (3.5 and 1.5 times higher than in non-smokers, respectively). Furthermore, smoking habits, such as number of smoked cigarettes per day, smoking period and cigarette type, along with age, were shown to influence these metals' blood concentration. Higher blood levels of Cd and Pb were found in smokers consuming more than 10 cigarettes per day for more than 10 years. The present study also highlighted the importance of the controlled tobacco production, since it was shown that consumption of illicit tobacco could manifold the exposure to toxic metals that can subsequently increase the frequency of related diseases as well.

Reduced hepatic metallothionein expression in first trimester fetuses in response to intrauterine smoking exposure: a consequence of low maternal zinc levels?

Does maternal smoking in early pregnancy affect metallothionein 1 and 2 (MT1 and MT2) mRNA and protein expression in first trimester placenta or embryonic/fetal liver? In the first trimester, MT protein expression is seen only in liver, where smoking is associated with a significantly reduced expression. Zinc homeostasis is altered by smoking. Smoking induces MT in the blood of smokers properly as a result of the cadmium binding capacities of MT. In term placenta MT is present and smoking induces gene and protein expression (MT2 in particular), but the MT presence and response to smoking have never been examined in first trimester placenta or embryonic/fetal tissues. Cross sectional study where the presence of MT mRNA and protein was examined at the time of the abortion. The material was collected with informed consent after surgical intervention and frozen immediately. For protein expression analysis, liver tissue originating from smoking exposed n = 10 and unexposed n = 12 pregnancies was used. For mRNA expression analyses, placental tissue originating from smokers n = 19 and non-smokers n = 23 and fetal liver tissue from smoking exposed n = 16 and smoking unexposed pregnancies n = 13, respectively, were used. Tissues were obtained from women who voluntarily and legally chose to terminate their pregnancy between gestational week 6 and 12. Western blot was used to determine the protein expression of MT, and real-time PCR was used to quantify the mRNA expression of MT2A and eight MT1 genes alongside the expression of key placental zinc transporters: zinc transporter protein-1 (ZNT1), Zrt-, Irt-related protein-8 and -14 (ZIP8 and ZIP14). A significant reduction in the protein expression of MT1/2 in liver tissue (P = 0.023) was found by western blot using antibodies detecting both MT forms. Overall, a similar tendency was observed on the mRNA level although not statistically significant. Protein expression was not present in placenta, but the mRNA regulation suggested a down regulation of MT as well. A suggested mechanism based on the known role of MT in zinc homeostasis could be that the findings reflect reduced levels of easily accessible zinc in the blood of pregnant smokers and hence a reduced MT response in smoking exposed fetal/embryonic tissues. Smoking was based on self-reports; however, our previous studies have shown high consistency regarding cotinine residues and smoking status. Passive smoking could interfere but was found mainly among smokers. The number of fetuses was limited, and other factors such as medication and alcohol might affect the findings. Information on alcohol was not consistently obtained, and we cannot exclude that it was more readily obtained from non-users. In the study, alcohol consumption was reported by a limited number (less than 1 out of 5) of women but with more smokers consuming alcohol. However, the alcohol consumption reported was typically limited to one or few times low doses. The interaction between alcohol and smoking is discussed in the paper. Notably we would have liked to measure zinc status to test our hypothesis, but maternal blood samples were not available. Zinc deficiency-in particular severe zinc deficiency-can affect pregnancy outcome and growth. Our findings indicate that zinc homeostasis is also affected in early pregnancy of smokers, and we know from pilot studies that even among women who want to keep their babies, the zinc status is low. Our findings support that zinc supplements should be considered in particular to women who smoke. We thank the Department of Biomedicine for providing laboratory facilities and laboratory technicians and the Lundbeck Foundation and Læge Sofus Carl Emil Friis og Hustru Olga Doris Friis Legat for financial support. The authors have no competing interests to declare. N/A.

Measuring the Concentration of Heavy Elements in Blood of Workers in Fuel Stations in Dhi Qar Governorate

This study was designed to measure the concentration of some heavy metals (Copper, Zinc, Iron and Lead) in blood serum of workers in filling stations in the center of Dhi Qar governorate in southern Iraq; Ten samples were selected from a group of workers ranging ages from (19 to 54 years), and had (1 to 11 years) work experience, some of them are smokers and others are not. The study included employment from Al-Raya and Mansouriya stations and also included people work at other places to be control for the study. Measurements were done in laboratories of the Faculty of Sciences, Dhi Qar University. Results statistically showed that concentrations of the four metals among workers in fuel stations are higher than those in the control, and the concentrations in blood of smokers are higher than in non- smokers workers, the study also confirmed that differences in ages percentages between workers groups where found.

Abstract 224: Sub-cytotoxic Levels of Heavy Metals Induce Pro-inflammatory Signaling in the Aortic Endothelium without Impairing Flow-Mediated Dilation in Rats

Introduction: Acute exposure to tobacco or marijuana secondhand smoke (SHS) causes endothelial dysfunction. The identity of specific SHS constituents that cause vascular toxicity is unclear. Heavy metals are present in SHS and at elevated levels in the blood of smokers, and may mediate acute endothelial dysfunction through reactive oxygen species formation and decreasing NO bioavailability. We assessed the effects of exposure to cadmium, lead, mercury, and arsenic at levels present in the blood of human smokers on flow-mediated dilation (FMD) as a measure of endothelial function in rats. We also evaluated the effects of heavy metal exposure on intimal structure, protein localization pattern, and inflammatory gene expression in the aortic endothelium. Hypothesis: Sub-cytotoxic levels of heavy metals impair FMD, alter intimal structure, and induce pro-inflammatory signaling in endothelium. Methods: We injected rats (n=8/group) with heavy metal cocktail or vehicle intravenously and quantitated pre- and post-exposure FMDs measures defined as percent vasodilation of femoral artery after transient ischemia. We performed en face aorta immunostaining and assessed endothelial cell axis alignment, cell length ratio, and localization pattern of PECAM-1, VE-cadherin, and vimentin. We also quantified gene expression of key endothelial proteins in aorta homogenates. Results: FMD was not impaired in the heavy metal group (8.8±3.6(SD)% vs. 12.9±8.0%, p=.31 or controls (7.5±2.7% vs. 8.8±5.8%, p=.63). No significant difference in cell length ratio and endothelial x and y axes of alignment were detected between groups (p&gt;.8) and localization of PECAM-1, VE-cadherin, and vimentin in the aorta endothelium remained unaltered following heavy metal injection. However, expression of PECAM-1 and VE-cadherin was significantly lower in the heavy metal-treated rats, while VCAM-1 gene expression was significantly higher (p&lt;.05). Conclusion: Acute exposure to sub-cytotoxic levels of heavy metals can induce pro-inflammatory signaling in endothelium, which could potentially lead to vascular injury. However, FMD and endothelial structure remain unchanged by heavy metal exposure.

Sampling and storage conditions for cannabinoid analysis: Plastic vs. glass

In forensic toxicology, analysed metabolites are THC-COOH, 11-OH-THC, Cannabidiol (CBD) and Cannabinol (CBN). However, cannabinoids are known to be adsorbed on plastics [1] . Cannabinoid stability is crucial for interpreting concentrations and actually, there are no studies that evaluate storage conditions in plastic tube versus glass tube. Furthermore, toxicology labs have to evaluate each pre analytical and analytical step in order to accreditation process due to ISO 15189 norms. In these conditions, we decided to evaluate sample storage in different sample type. THC -COOH, 11-OH-THC, CBD and CBN were spiked in different concentrations to urine drug free, plasma drug free and whole blood drug free. Concentrations were chosed following concentrations found in light and heavy cannabis smoker's blood sample. Spiked samples were then aliquoted. In half, we added antioxidant to prevent oxidation degradation [2] . Each condition was then stored at +4 °C, −20 °C and −80 °C and aliquots were analysed at different times until 6 months (n = 5). Each condition was analysed by screening urinary test, confirmation analysis by biochips with Whole Blood and Urinary Drug Of Abuse kit (Randox ® Investigator) and by a specific analysis on UPLC-MS/MS (Waters Acquity TQS ® ). Whatever analytical methods, we didn’t find any significative difference between glass tubes and plastic tubes concentrations at −20 °C and −80 °C during first months. Storage stability was judged acceptable after 1 month at −20 °C and −80 °C. Latest stability results are occurring and will be shown in case of acceptation of the submission. Furthermore, no degradation/adsorption was noticed in the first week storage at +4 °C. This study showed storage stability conditions until 6 months. We also demonstrated that +4 °C stability is compliant with analysis delay in toxicology lab. Plastic tube and glass tube didn’t shown significative difference in term of adsorption/degradation. We can conclude that cannabinoid test sample can be collected and stored in plastic tubes and don’t need specific storage. This conclusion is also a major advantages concerning conservatory toxicology sampling.

Detoxifying of Nicotine in Smoker with Consumption of Food Containing CYP 2A6 Enzyme from Beef Liver

Background: the addicted smoking nicotine levels in the blood have an average 12 mg/liter of blood, while the safe limit of nicotine in the blood is 2 mg/liter. Nicotine levels decreased can be done by using the food contains biotransformation enzymes i.e. CYP 2A6. The purpose of this study was to know the decrease in nicotine in the blood of smokers after the feeding of biotransformation enzyme containing nicotine CYP 2A6 Method: The research method was an experiment. The number of respondents active smokers 30 people which received rehabilitation at PHC Surabaya Hospital . Samples were taken by random sampling. Result: Based on the results of the study, second inspection after consumption of food which contained enzyme CYP 2A6 from beef liver, there was an increased excretion of cotinine in urine. There were 80% of respondents have increased levels excretion of cotinine in urine. Conclusion: The conclusion of this study was that food which contained nicotine biotransformation enzyme (CYP 2A6) decreased concentration of nicotine in the blood of smokers

Abstract B14: Modulating effects of green and black tea on biomarkers of chronic inflammation by gender and smoking status

Abstract Background/Purpose: Epidemiologic data implies that there are gender differences in lung cancer pathogenesis and possibly increased susceptibility to lung cancer in women. Chronic inflammation has been implicated as important modulator of human health by playing a significant role in both disease prevention and disease development. Several studies have demonstrated increased interleukin 6 (IL-6) and C-reactive protein (CRP) in the blood of smokers. The overall goal of this study was to develop a feasible tea intervention that will serve as a model for the chemoprevention of a wide range of tobacco-related diseases. Our immediate goal, that was addressed over a 4-year study period, was to determine the effects of high tea consumption on biological markers of chronic inflammation that mediate lung cancer risk, including, IL-6, CRP and antioxidant enzyme activities. Methods: We completed a 6-month randomized, controlled, double-blinded trial in a group of current and former smokers. Participants were stratified on smoking status and gender, and were randomized to green or black tea preparations or a control intervention (matching placebo). Levels of urinary iIL-6 and CRP are used to measure chronic inflammation and levels of superoxide dismutase (SOD) in red blood cells are used to measure antioxidant enzymes. Results: The study protocol was approved by all parties. A total of 138 participants (78 females and 60 males) completed the study. Our data show that il6 is significantly correlated with years of smoking and pack/year among smokers and former smokers. At the end of the 6-month intervention, female smokers in the green tea group showed a significant decrease in IL-6 (P=0.036) while male former smokers in the black tea group showed a significant decrease in CRP levels (P=0.012). There were no significant changes in dietary and serum antioxidant levels between the different groups. Conclusion: This data implies that smokers are more likely to benefit from green tea intake while the beneficial effects of black tea are observed among former smokers. Note: This abstract was not presented at the conference. Citation Format: Iman A. Hakim, Sami A. Aldaham, Janet Foote, H-H Sherry Chow. Modulating effects of green and black tea on biomarkers of chronic inflammation by gender and smoking status. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr B14.