Chronic Obstructive Pulmonary Disease: Genetic Polymorphisms and Intermediary Metabolism Alterations

Chronic obstructive pulmonary disease (COPD) is a multifactorial disease characterized by airflow obstruction that is usually progressive and associated with an abnormal inflammatory response of the lungs to noxious particles and gases. Cigarette smoke (CS) is the main risk factor, but only a small proportion of smokers (15-20%) develop symptomatic disease, this suggests that there are individual susceptibility factors involved in disease onset and progression. Considering the impact of environmental and genetic risk factors in COPD, this dissertation sought to uncover genetic susceptibility biomarkers in a population affected by COPD and explore tissue metabolic alterations induced by chronic CS exposure in a mouse model. A case-control study was carried on in a COPD population, aiming to investigate whether polymorphisms of microsomal epoxide hydrolase (EPHX1) gene, and related phenotypes, had any bearing on individual susceptibility to COPD onset and severity. DNA of COPD patients and controls was genotyped for functional polymorphisms of EPHX1 gene (exon 3 Tyr113His, exon 4 His139Arg), and haplotype analysis was performed using PCR-RFLP and PCR-RT techniques. The statistical analysis did not show any significant result about the potential relationship between analyzed SNPs, related phenotypes, and COPD risk and severity. Stable isotope-resolved metabolomics approach was used to study glycolytic pathway alterations induced by chronic CS exposure in lung and liver tissue of an emphysema mouse model. C57BL/6J mice, after CS exposure, were injected via IP injection with glucose tracer containing carbon stable isotope - 13C6-glucose – then, tissues were collected and the incorporation of 13C into metabolites was monitored by mass spectrometry and nuclear magnetic resonance spettroscopy. Lung tissue analyses did not reveal any significant alteration in lung tissue glycolysis of mice exposed to CS. On the other hand, CS may contribute to dysregulated glycolysis, PPP, glycogen synthesis and utilization, in emphysema mouse model liver tissue

Exposure to electronic cigarettes impairs pulmonary anti-bacterial and anti-viral defenses in a mouse model

© 2015 Sussan et al. Electronic cigarettes (E-cigs) have experienced sharp increases in popularity over the past five years due to many factors, including aggressive marketing, increased restrictions on conventional cigarettes, and a perception that E-cigs are healthy alternatives to cigarettes. Despite this perception, studies on health effects in humans are extremely limited and in vivo animal models have not been generated. Presently, we determined that E-cig vapor contains 7x1011 free radicals per puff. To determine whether E-cig exposure impacts pulmonary responses in mice, we developed an inhalation chamber for E-cig exposure. Mice that were exposed to E-cig vapor contained serum cotinine concentrations that are comparable to human E-cig users. E-cig exposure for 2 weeks produced a significant increase in oxidative stress and moderate macrophage-mediated inflammation. Since, COPD patients are susceptible to bacterial and viral infections, we tested effects of E-cigs on immune response. Mice that were exposed to E-cig vapor showed significantly impaired pulmonary bacterial clearance, compared to air-exposed mice, following an intranasal infection with Streptococcus pneumonia. This defective bacterial clearance was partially due to reduced phagocytosis by alveolar macrophages from E-cig exposed mice. In response to Influenza A virus infection, E-cig exposed mice displayed increased lung viral titers and enhanced virus-induced illness and mortality. In summary, this study reports a murine model of E-cig exposure and demonstrates that E-cig exposure elicits impaired pulmonary anti-microbial defenses. Hence, E-cig exposure as an alternative to cigarette smoking must be rigorously tested in users for their effects on immune response and susceptibility to bacterial and viral infections

Relationship between lipid phenotypes, overweight, lipid lowering drug response and KIF6 and HMG-CoA genotypes in a subset of the brisighella heart study population

The existence of genetic traits might explain the susceptibility to develop hypercholesterolemia and the inter-individual differences in statin response. This study was performed to evaluate whether individuals\ue2\u80\u99 polymorphisms in HMG-CoA and KIF6 genes are independently associated with hypercholesterolemia, other lipid-associated traits, and statin response in unselected individuals enrolled in the Brisighella heart study (Survey 2012). A total of 1622 individuals, of which 183 under statin medication, were genotyped for a total of five polymorphisms (KIF6 rs20455, rs9471077, rs9462535, HMG-CoA rs3761740, rs3846662). The relationships between the five loci and clinical characteristics were analyzed. The principal basic parameters calculated on 12 h fasting blood included total cholesterol (TC), High Density Lipoprotein Cholesterol (HDL-C), Low-Density Lipoprotein Cholesterol (LDL-C), and triglycerides (TG). Hypercholesterolemia was defined as a TC >200 mg/dL or use of lipid-lowering medication. 965 individuals were characterized by hypercholesterolemia, these subjects were significantly older (p < 0.001), with body mass index (BMI) and waist circumference significantly higher (p < 0.001) compared to the others. HMG-CoA rs3846662 GG genotype was significantly over-represented in the hypercholesterolemic group (p = 0.030). HMG-CoA rs3846662 genotype was associated with the level of TC and LDL-C. Furthermore, in the same subset of untreated subjects, we observed a significant correlation between the KIF6 rs20455 and HDL-C. KIF6 variants were associated with a significantly lower (rs20455) or higher (rs9471077 and rs9462535) risk of obesity, in males only. No association between responsiveness to statins and the polymorphisms under investigation were observed. Our results showed associations between HMG-CoA rs3846662 and KIF6 rs20455 and lipid phenotypes, which may have an influence on dyslipidemia-related events. Moreover, this represents the first study implicating KIF6 variants with obesity in men, and point to the possible involvement of this genetic locus in the known gender-related differences in coronary artery disease

Relationship between Lipid Phenotypes, Overweight, Lipid Lowering Drug Response and KIF6 and HMG-CoA Genotypes in a Subset of the Brisighella Heart Study Population

The existence of genetic traits might explain the susceptibility to develop hypercholesterolemia and the inter-individual differences in statin response. This study was performed to evaluate whether individuals’ polymorphisms in HMG-CoA and KIF6 genes are independently associated with hypercholesterolemia, other lipid-associated traits, and statin response in unselected individuals enrolled in the Brisighella heart study (Survey 2012). A total of 1622 individuals, of which 183 under statin medication, were genotyped for a total of five polymorphisms (KIF6 rs20455, rs9471077, rs9462535; HMG-CoA rs3761740, rs3846662). The relationships between the five loci and clinical characteristics were analyzed. The principal basic parameters calculated on 12 h fasting blood included total cholesterol (TC), High Density Lipoprotein Cholesterol (HDL-C), Low-Density Lipoprotein Cholesterol (LDL-C), and triglycerides (TG). Hypercholesterolemia was defined as a TC &gt;200 mg/dL or use of lipid-lowering medication. 965 individuals were characterized by hypercholesterolemia; these subjects were significantly older (p &lt; 0.001), with body mass index (BMI) and waist circumference significantly higher (p &lt; 0.001) compared to the others. HMG-CoA rs3846662 GG genotype was significantly over-represented in the hypercholesterolemic group (p = 0.030). HMG-CoA rs3846662 genotype was associated with the level of TC and LDL-C. Furthermore, in the same subset of untreated subjects, we observed a significant correlation between the KIF6 rs20455 and HDL-C. KIF6 variants were associated with a significantly lower (rs20455) or higher (rs9471077 and rs9462535) risk of obesity, in males only. No association between responsiveness to statins and the polymorphisms under investigation were observed. Our results showed associations between HMG-CoA rs3846662 and KIF6 rs20455 and lipid phenotypes, which may have an influence on dyslipidemia-related events. Moreover, this represents the first study implicating KIF6 variants with obesity in men, and point to the possible involvement of this genetic locus in the known gender-related differences in coronary artery disease

Schematic of our E-cig exposure model.

<p>Schematic of our E-cig exposure model.</p

E-cig exposure reduces pulmonary bacterial clearance in mice infected with <i>S</i>. <i>pneumoniae</i>.

<p>Mice were exposed to air or E-cig for 2 wks, then infected intranasally with 1x10⁵ colony forming units (CFU) of <i>S</i>. <i>pneumoniae</i>. Inflammation (A) and bacterial CFUs (B) were determined in BAL at 24h after infection (N = 10 mice per group). (C) In a separate group of mice, bacterial CFUs were quantified in lung homogenates (N = 10 mice per group). (D) Alveolar macrophages from air or E-cig exposed mice were harvested and infected with <i>S</i>. <i>pneumoniae</i> at multiplicities of infection of 10 and 20. Bacterial CFUs were quantified in cell-free culture media at 4 h after infection. (E) Intracellular (internalized) and extracellular bacteria (cell-free culture media) were quantified at 1 h after <i>ex vivo</i> infection of alveolar macrophages with an MOI of 20. (F) Alveolar macrophages were harvested at 2h after final exposure, stained with antibodies against CD36 and MARCO, and analyzed by flow cytometry. (G) Bacterial clearance was measured in alveolar macrophages from mice exposed to air or traditional E-cig vapor. *p<0.05 by Student’s two-tailed t-test.</p

E-cig-induced pulmonary response.

<p>(A) EPR spectra of E-cig TPM. The presented spectrum is a result of subtraction of a Cambridge filter pad EPR signal before and after collection of TPM. (B) Lipid peroxidation was measured by thiobarbituric acid reactive substances (TBARS) in lung homogenates from C57BL/6 mice that were exposed to air or E-cig vapor for 1.5 h, twice per day, for 2 wks. (C) Inflammatory cells were quantified in the BAL at 24h after the final exposure. (D) Cytokines were analyzed in cell-free BAL fluid from air and E-cig exposed mice at 24h after the final exposure. N = 10 mice per group. *p<0.05 by Student’s two-tailed t-test.</p

E-cig exposure impairs viral clearance and causes significant morbidity and mortality in mice following influenza virus infection.

<p>Mice were exposed to air or E-cig for 2 wks, then infected intranasally with either TCID₅₀ 10² (A-B) or TCID₅₀ 10³ (C) of H1N1 virus. (A) Viral titer was determined by TCID₅₀ assay in lung homogenates at 4 days after infection (N = 5 mice per group). (B-C) Mice were weighed daily after infection with either TCID₅₀ 10² (B) or 10³ (C), and values are presented as percent of starting weight (N = 10 mice per group). For mice that died during the experiments, body weights were included in the analysis up until the day of death. (D) Mortality curves in response to intranasal infection with TCID₅₀ 10² or 10³ H1N1 (N = 10 mice per group). *p<0.05 by Student’s two-tailed t-test.</p

Influenza-induced inflammation is altered by E-cig exposure.

<p>Mice were exposed to air or E-cig for 2 wks, then infected intranasally with TCID₅₀ 10² of H1N1. BAL was collected at day 4 (N = 5) and day 8 (N = 4) after infection, followed by quantification of inflammatory cells (A) and cytokines (B). *p<0.05 by Student’s two-tailed t-test.</p

CNR-DT 210/2013 - Istruzioni per la Progettazione, l'Esecuzione ed il Controllo delle Strutture di Vetro

Gli sviluppi tecnologici hanno consentito negli ultimi decenni una strabiliante espansione delle applicazioni del vetro nel settore delle costruzioni. In virt\uf9 della sua trasparenza o traslucenza, questo materiale, che caratterizza alcune tendenze dell\u2019architettura moderna, vede moltiplicare le sue applicazioni in forma di pannelli di grande superficie, coperture, solai, scale, pareti, pilastri, parapetti. Gli elementi di vetro, utilizzati inizialmente con semplice funzione di tamponamento o rivestimento, costituiscono oggi strutture vere e proprie che, di conseguenza, devono essere sottoposte a procedure di calcolo, valutazione e controllo, dello stesso tipo di quelle utilizzate per tutti i materiali strutturali. La funzione strutturale \ue8 quindi nuova per un materiale antico, ma richiede particolare attenzione nel dimensionamento e nell\u2019impiego. Costruire con il vetro piuttosto che con altri materiali non \ue8 n\ue9 pi\uf9 difficile n\ue9 pi\uf9 complesso, ma richiede la considerazione di aspetti specifici essenzialmente legati alla sua fragilit\ue0 intrinseca. Un approccio consapevole alla progettazione pu\uf2 portare a soluzioni tecniche che consentono, in ogni caso, il raggiungimento di livelli di affidabilit\ue0 e sicurezza analoghi a quelli ottenibili nelle opere di costruzione che utilizzano materiali strutturali pi\uf9 tradizionali, come ad esempio il calcestruzzo o l\u2019acciaio