Downregulation of lung mitochondrial prohibitin in COPD

Summary Prohibitins (PHB1 and PHB2) are versatile proteins located at the inner mitochondrial membrane, maintaining normal mitochondrial function and morphology. They interact with the NADH dehydrogenase protein complex, which is essential for oxidoreductase activity within cells. However, their expression in lung epithelium, especially in smokers and patients with inflammatory lung diseases associated with increased oxidative stress, such as COPD, is unknown. Lung tissue specimens from 45 male subjects were studied: 20 COPD patients [age: 65.7 AE 5.8 years, smoking: 84.6 AE 33.6 pack-years, FEV 1 (%pred.): 58.7 AE 14.6, FEV 1 /FVC (%): 63.8 AE 9.4], 15 non-COPD smokers [age: 59.0 AE 12.1 years, smoking: 52.5 AE 20.8 pack-years, FEV 1 (%pred.): 85.5 AE 14.2, FEV 1 /FVC (%): 78.5 AE 4.7] and 10 non-smokers. Quantitative real-time PCR experiments were carried out for PHB1 and PHB2, using b-actin as internal control. Non-COPD smokers exhibited lower PHB1 mRNA levels when compared to non-smokers (0.55 AE 0.06 vs. 0.90 AE 0.06, P Z 0.043), while PHB1 expression was even further decreased in COPD patients (0.32 AE 0.02), a statistically significant finding vs. both non-COPD smokers (P Z 0.040) and non-smokers (P < 0.001). By contrast, PHB2 levels were similar among the three study groups. Western blot analysis for the PHB1 protein verified the qPCR results (non-smokers: 1.77 AE 0.13; non-COPD smokers: 0.97 AE 0.08; COPD patients: 0.59 AE 0.10, P Z 0.007). Further analysis revealed that PHB1 downregulation in COPD patients cannot be attributed solely to smoking, and that PHB1 expression levels are associated with the degree of airway obstruction [FEV 1 (P mRNA Z 0.004, P protein Z 0.014)]. The significant downregulation of PHB1 in COPD and non-COPD smokers in comparison to non-smokers possibly reflects a distorted mitochondrial function due to decreased mitochondrial stability, especially in the mitochondria of COPD patients.

DNA Damage Due to Oxidative Stress in Chronic Obstructive Pulmonary Disease (COPD)

According to the American Thorasic Society (ATS)/European Respiratory Society (ERS) Statement, chronic obstructive pulmonary disease (COPD) is defined as a preventable and treatable disease with a strong genetic component, characterized by airflow limitation that is not fully reversible, but is usually progressive and associated with an enhanced inflammatory response of the lung to noxious particles or gases. The main features of COPD are chronic inflammation of the airways and progressive destruction of lung parenchyma and alveolar structure. The pathogenesis of COPD is complex due to the interactions of several mechanisms, such as inflammation, proteolytic/antiproteolytic imbalance, oxidative stress, DNA damage, apoptosis, enhanced senescence of the structural cells and defective repair processes. This review focuses on the effects of oxidative DNA damage and the consequent immune responses in COPD. In susceptible individuals, cigarette smoke injures the airway epithelium generating the release of endogenous intracellular molecules or danger-associated molecular patterns from stressed or dying cells. These signals are captured by antigen presenting cells and are transferred to the lymphoid tissue, generating an adaptive immune response and enhancing chronic inflammation

The implication of herpes viruses in male infertility

Infertility accounts for one of the problems the affect almost 15-20% of the couples in European countries. 30% of the patients suffer from idiopathic infertility. Raising evidence suggest that viral infections contribute to male infertility. Viral DNA from the members of the Herpesviridae family (HSV-1, HSV-2, CMV or EBV), have been detected in the semen of asymptomatic sterile patients. The aim of this PhD thesis was the detection of the 8 members of the Herpesviridae family in semen samples and the correlation of their presence or no with the samples’ altered semen parameters. Firstly, fresh semen samples were collected and characterized based on the spermogram. Secondly, the detection experiments for the herpes viruses were performed by PCR. Thirdly, the presence or no of one or more viruses and the parameters of the samples in each sample group were correlated and a statistical analysis was performed. In vivo experiments were also conducted in order to confirm the capacity of infection of CMV in spermatozoa. Semen samples were incubated in culture medium with the virus. The statistical analysis revealed very interesting results. The semen showed high viral presence (92,8%) independently of the characterization as normal or abnormal. Interestingly, there was a statistical significance between the EBV presence and the pyospermia, in all samples (p=0,001). The presence of EBV influences the concentration of WBCs detected in semen and justifies the statistical significance observed between the EBV presence and the normal samples with increased WBCs. The HHV-6 presence was statistically significant (p=0,001), which according to WHO are evidence of immunologic infertility. These results suggest that HHV-6 affects those semen parameters concerning seminal fluid. The high presence of HHV-8 was statistically significant in samples with oligoasthenozoospermia. A statistical significant correlation was also found between samples positive for HHV-8 and reduced concentration of spermatozoa (p=0,030) and reduced motility (p=0,015). Concluding, the presence of HHV-8 in sperm samples seems to reduce the mean number and motility of spermatozoa, causing oligoasthenozoospermia. The results of this PhD Thesis suggest the implication of herpes viruses in altered semen parameters, while their detection could be incorporated in the routine semen analysis together with the spermogram and applied both to patients seeking fertility evaluation and those performing IVFΗ υπογονιμότητα αποτελεί ένα από τα προβλήματα που επηρεάζουν σχεδόν το 15-20% των ζευγαριών στις Ευρωπαϊκές χώρες. Tο 30% των ασθενών υποφέρουν από ιδιοπαθή υπογονιμότητα. Ολοένα και περισσότερες ενδείξεις υποδεικνύουν ότι οι ιϊκές μολύνσεις συμβάλλουν στην ανδρική υπογονιμότητα. Ιϊκό DNA από μέλη της οικογένειας Herpesviridae, όπως οι HSV-1, HSV-2, CMV ή EBV έχουν ανιχνευτεί στο σπέρμα ασυμπτωματικών στείρων ασθενών. Σκοπός της παρούσας διατριβής ήταν η ανίχνευση των 8 μελών της οικογένειας των ερπητοϊών (HSV1&2, VZV, EBV, CMV, HHV-6, HHV-7 και HHV-8) σε δείγματα σπέρματος και η συσχέτιση της παρουσίας τους ή μη με τις παραμέτρους του σπέρματος. Στο πρώτο μέρος της διατριβής έγινε συλλογή φρέσκων δειγμάτων σπέρματος και χαρακτηρισμός τους βάση σπερμοδιαγράμματος. Πραγματοποιήθηκαν πειράματα ανίχνευσης του DNA των μελών της οικογένειας των ερπητοϊών με τη μέθοδο της PCR. Στο τρίτο μέρος, πραγματοποιήθηκε η συσχέτιση των αποτελεσμάτων και η διεξαγωγή στατιστικής ανάλυσης. Από την στατιστική ανάλυση τα αποτελέσματα υποδεικνύουν υψηλή παρουσία όλων των ερπητοϊών (92,8%) στο σπέρμα ανεξάρτητα από την κατηγοριοποίηση σε φυσιολογικά και μη- φυσιολογικά δείγματα. Ιδιαίτερα σημαντική είναι η στατιστική σημαντικότητα που εμφανίστηκε ανάμεσα στην παρουσία του EBV και στα δείγματα με πυοσπερμία. Η παρουσία του EBV επηρεάζει τον αριθμό των WBCs που ανιχνεύονται στο σπέρμα, καθιστώντας το πυοσπερμικό. Η παρουσία του HHV-6 ήταν στατιστικά σημαντική (p=0,001) στα δείγματα με φυσιολογικά μεν σπερματοζωάρια παθολογικό δε σπερματικό υγρό και/ή παρουσία υψηλής συγκέντρωσης WBCs και/ή συγκόλληση σπερματοζωαρίων. Τα παραπάνω τροποποιημένα χαρακτηριστικά αποτελούν ένδειξη ανοσολογικής υπογονιμότητας σύμφωνα με τον Π.Ο.Υ. Η υψηλή παρουσία του HHV-8 ήταν στατιστικά σημαντική στα δείγματα με ολιγοασθενοζωοσπερμία. Η παρουσία του ιού HHV-8 στα δείγματα σπέρματος φαίνεται ότι μειώνει το μέσο αριθμό και την κινητικότητα των σπερματοζωαρίων, προκαλώντας έτσι εμφάνιση ολιγοασθενοζωοσπερμίας. Στο τελευταίο μέρος πραγματοποιήθηκαν πειράματα επιβεβαίωσης της ικανότητας μόλυνσης των σπερματοζωαρίων από τον ΗCMV. Τα αποτελέσματα της παρούσης διατριβής παρέχουν σοβαρές ενδείξεις για τη εμπλοκή των ερπητοϊών στις τροποποιημένες παραμέτρους του σπέρματος, ενώ θα μπορούσε να αξιοποιηθεί η ανίχνευση τους στον έλεγχο ρουτίνας μαζί με το σπερμοδιάγραμμα σε ασθενείς που αποζητούν τον έλεγχο της γονιμότητας τους αλλά και σε εκείνους που επιθυμούν εξωσωματική γονιμοποίησ

DNA Damage Due to Oxidative Stress in Chronic Obstructive Pulmonary Disease (COPD)

Society (ERS) Statement, chronic obstructive pulmonary disease (COPD) is defined as a preventable and treatable disease with a strong genetic component, characterized by airflow limitation that is not fully reversible, but is usually progressive and associated with an enhanced inflammatory response of the lung to noxious particles or gases. The main features of COPD are chronic inflammation of the airways and progressive destruction of lung parenchyma and alveolar structure. The pathogenesis of COPD is complex due to the interactions of several mechanisms, such as inflammation, proteolytic/antiproteolytic imbalance, oxidative stress, DNA damage, apoptosis, enhanced senescence of the structural cells and defective repair processes. This review focuses on the effects of oxidative DNA damage and the consequent immune responses in COPD. In susceptible individuals, cigarette smoke injures the airway epithelium generating the release of endogenous intracellular molecules or danger-associated molecular patterns from stressed or dying cells. These signals are captured by antigen presenting cells and are transferred to the lymphoi

Polymorphisms of fractalkine receptor CX3CR1 gene in patients with symptomatic and asymptomatic carotid artery stenosis

Aim: The chemokine fractalikine is expresses in vascular endothelium, exerting a pro-atherogenic effect. Two single-nucleotide polymorphisms of the CX3CR1 gene (T280M and V2491) affect fractalkine receptor expression and function. We aimed to assess the prevalence of CX3CR1 polymorphisms and the asociation with ischemic cerebrovascular attacts in a cohort of carotid atheromatous disease patients and age-matched controls. Methods: Using PCR-RFLP, we analyzed allelotypes for T280M and V249I in 150 patients with and 151 controls without carotid atherosclerosis assessed using carotid duplex ultrasound; the sugjects were patients admitted for any reason to a tertiary hospital. Genotype data were compared with modifiable risk factors for cerebrovascular disease and the reason for admission, using ischemic stroke as an endpoint. Stroke types associated with carotid atherosclerosis were analysed separately. Results: The M280 allelic frequency was lower among carotid atherosclerosis patients than controls (0.15 versus 0.23, adjusted OR 0.47, 95% CI 0.30-0.74). Absence of M280 allele was an independent factor associated with carotid atherosclerosis (OR 3.70, 95% CI 1.92-7.14), strongers than hypertension, dyslipidemia, diabetes and cigarette smoking. The I249 allele was also under-represented in carotid atherosclerosis; this was not statistically significant. T280M and V249I genotypes were not associated with admission due to ischemic stroke of the large vessel subtype (TOAST classification, 73 episodes), whereas carotid atherosclerosis, previous ischemic event, age, hypertension, diabetes, hyperlipidemia and cigarette smoking were all independently associated. Conclusions: The M280 fractalkine receptor gene allele is associated with a lower risk of carotid atheromatous disease, independent from the modifiable cerebrovascular risk factors