Extracellular Vesicles in Biological Fluids. A Biomarker of Exposure to Cigarette Smoke and Treatment with Chemopreventive drugs

Extracellular vesicles (EVs) are released from cells and enter into body fluids thereby providing a toxicological mechanism of cell-cell communication. The present study aimed at assessing (a) the presence of EVs in mouse body fluids under physiological conditions, (b) the effect of exposure of mice to cigarette smoke for 8 weeks, and (c) modulation of smoke-related alterations by the nonsteroidal anti-inflammatory drug celecoxib, a selective cyclooxygenase-2 inhibitor. To this purpose, ICR (CD-1) mice were either unexposed or exposed to cigarette smoke, either treated or untreated with oral celecoxib. EVs, isolated from bronchoalveolar lavage fluid (BALF), blood serum, and urines, were analyzed by nanoparticle tracking analysis and flow cytometry. EVs baseline concentrations in BALF were remarkably high. Larger EVs were detected in urines. Smoking increased EVs concentrations but only in BALF. Celecoxib remarkably increased EVs concentrations in the blood serum of both male and female smoking mice. The concentration of EVs positive for EpCAM, a mediator of cell-cell adhesion in epithelia playing a role in tumorigenesis, was much higher in urines than in BALF, and celecoxib significantly decreased their concentration. Thus, the effects of smoke on EVs concentrations were well detectable in the extracellular environment of the respiratory tract, where they could behave as delivery carriers to target cells. Celecoxib exerted both protective mechanisms in the urinary tract and adverse systemic effects of likely hepatotoxic origin in smoke-exposed mice. Detection of EVs in body fluids may provide an early diagnostic tool and an end-point exploitable for preventive medicine strategies.

Contribution of HOGG1 Ser326Cys Polymorphism to the Development of Prostate Cancer in Smokers: Meta-Analysis of 2779 Cases and 3484 Controls

The HOGG1 gene catalyzes the excision of modified bases and removal of DNA damage adducts. It may play an important role in the prevention of carcinogenesis. Ser326Cys polymorphism localizes in exon 7 of the hOGG1 gene. It takes the form of an amino acid substitution, from serine to cysteine, in codon 326. Several epidemiological association studies have been conducted on this polymorphism and its relationship with the risk of prostate cancer. However, results have been conflicting. To resolve this conflict, we conducted a meta-analysis on the association between this polymorphism and prostate cancer, taking into account race, country, sources of controls, and smoking status. A total of nine studies covering 2779 cases and 3484 controls were included in the current meta-analysis. Although no significant association was found between hOGG1 Ser326Cys polymorphism and prostate cancer susceptibility in the pooled analysis, individuals with Ser/Cys+Cys/Cys genotypes were found to have greater risk of prostate cancer if they were also smokers (OR = 2.66, 95% CI = 1.58−4.47) rather than non-smokers (OR = 2.18, 95% CI = 1.13−4.19), compared with those with Ser/Ser genotype. In conclusion, our meta-analysis demonstrates that hOGG1 Ser326Cys polymorphism is a risk factor for prostate cancer in smokers. Further studies are needed to confirm this relationship

Does clinical examination aid in the diagnosis of urinary tract infections in women? A systematic review and meta-analysis

<p>Abstract</p> <p>Background</p> <p>Clinicians should be aware of the diagnostic values of various symptoms, signs and antecedents. This information is particularly important in primary care settings, where sophisticated diagnostic approaches are not always feasible. The aim of the study is to determine the probability that various symptoms, signs, antecedents and tests predict urinary tract infection (UTI) in women.</p> <p>Methods</p> <p>We conducted a systematic search of the MEDLINE and EMBASE databases to identify articles published in all languages through until December 2008. We particularly focused on studies that examined the diagnostic accuracy of at least one symptom, sign or patient antecedent related to the urinary tract. We included studies where urine culture, a gold standard, was preformed by primary care providers on female subjects aged at least 14 years. A meta-analysis of the likelihood ratio was performed to assess variables related to the urinary tract symptoms.</p> <p>Results</p> <p>Of the 1, 212 articles identified, 11 met the selection criteria. Dysuria, urgency, nocturia, sexual activity and urgency with dysuria were weak predictors of urinary tract infection, whereas increases in vaginal discharge and suprapubic pain were weak predictors of the absence of infection. Nitrites or leukocytes in the dipstick test are the only findings that clearly favored a diagnosis of UTI.</p> <p>Conclusions</p> <p>Clinical findings do not aid in the diagnosis of UTI among women who present with urinary symptoms. Vaginal discharge is a weak indicator of the absence of infection. The urine dipstick test was the most reliable tool for detecting UTI.</p

Phosphodiesterase-4 Inhibition Alters Gene Expression and Improves Isoniazid – Mediated Clearance of Mycobacterium tuberculosis in Rabbit Lungs

Tuberculosis (TB) treatment is hampered by the long duration of antibiotic therapy required to achieve cure. This indolent response has been partly attributed to the ability of subpopulations of less metabolically active Mycobacterium tuberculosis (Mtb) to withstand killing by current anti-TB drugs. We have used immune modulation with a phosphodiesterase-4 (PDE4) inhibitor, CC-3052, that reduces tumor necrosis factor alpha (TNF-α) production by increasing intracellular cAMP in macrophages, to examine the crosstalk between host and pathogen in rabbits with pulmonary TB during treatment with isoniazid (INH). Based on DNA microarray, changes in host gene expression during CC-3052 treatment of Mtb infected rabbits support a link between PDE4 inhibition and specific down-regulation of the innate immune response. The overall pattern of host gene expression in the lungs of infected rabbits treated with CC-3052, compared to untreated rabbits, was similar to that described in vitro in resting Mtb infected macrophages, suggesting suboptimal macrophage activation. These alterations in host immunity were associated with corresponding down-regulation of a number of Mtb genes that have been associated with a metabolic shift towards dormancy. Moreover, treatment with CC-3052 and INH resulted in reduced expression of those genes associated with the bacterial response to INH. Importantly, CC-3052 treatment of infected rabbits was associated with reduced ability of Mtb to withstand INH killing, shown by improved bacillary clearance, from the lungs of co-treated animals compared to rabbits treated with INH alone. The results of our study suggest that changes in Mtb gene expression, in response to changes in the host immune response, can alter the responsiveness of the bacteria to antimicrobial agents. These findings provide a basis for exploring the potential use of adjunctive immune modulation with PDE4 inhibitors to enhance the efficacy of existing anti-TB treatment

Growth and decline of the COVID-19 epidemic wave in Italy from March to June 2020

March 21, 2020 was the ridgeline between the growth of the coronavirus disease 2019 (COVID-19) epidemic wave in Italy and the start of its decline. We analyzed the epidemic patterns from March 1 to June 30. There was a progressive drop of cases from March (104,710) to April (94,888), May (25,705) and June (8110). Likewise, after a slight increase of deaths in April (14,804) compared to March (12,396), a considerable decline occurred in May (5170) and June (1464). Doubling times of cumulative cases grew from 2 to 6 days until March 20 to 2 weeks up to April 5, and thereafter no further doubling occurred until June 30. There was a striking North\u2013South gradient of both cases and deaths. At the end of June, the nine Northern Italian regions or provinces, five central regions, and seven southern regions had contributed to the 81.1%, 12.4%, and 6.5% of the 240,578 national cases, respectively. Lombardy, the most populous region, was by far the most heavily affected one, accounting for the 39.0% of the national cases occurring over the analyzed 4-month period. However, in relative terms, it was preceded by Aosta Valley, the least populous region, less than 1% of the population of both regions having been affected by cases of COVID-19. The curves showing the ratio of daily cumulative cases and deaths to those of the previous day tended to flatten with time by approaching the zero growth but without reaching it, which documents a persisting circulation of severe acute respiratory syndrome coronavirus 2 in the Italian territory

MEDICINA PREVENTIVA E MEDICINA PREDITTIVA. NUOVE FRONTIERE PER L\u2019IGIENISTA

OBIETTIVI: L\u2019insorgere della medicina molecolare mette oggi a disposizione del medico preventivo nuovi strumenti rivolti alla valutazione del rischio di malattia cronico-degenerativa nel soggetto ancora sano. Tale valutazione pu\uf2 essere effettuata tramite indagine a livello del DNA o a valle dell\u2019espressione genica (indagini postgenomiche). METODI: Le indagine sul DNA valutano il rischio endogeno di patologia conferito dall\u2019assetto genetico polimorfico individuale. Tali indagini possono essere rivolte alla ricerca di geni ad elevata penetranza predittivi di patologia ma a bassa frequenza nella popolazione. In questo caso il loro ruolo in Sanit\ue0 Pubblica \ue8 piuttosto limitato e l\u2019esigenza di effettuare screening genetici di questo tipo non giustificata in termini di ricaduta sulla Salute Pubblica. I geni polimorfici molto diffusi nella popolazione sono invece scarsamente predittivi di patologia; anche in questo caso quindi lo procedura di screening non appare giustificata. Quindi il rischio di patologia su base puramente endogena \ue8 di rilevanza minoritaria rispetto al rischio esogeno di origine ambientale. RISULTATI: L\u2019indagine molecolare predittiva deve quindi avvalersi di metodiche tese ad identificare nel soggetto sano l\u2019esito della interazione tra fattori di rischio ambientali ed endogeni. Allo scopo pu\uf2 essere valutata la presenza di molecole genotossiche nel DNA. Tuttavia la predittivit\ue0 dell\u2019indagine aumenta quando l\u2019indagine si sposta verso l\u2019end point fenotipico. Pertanto l\u2019analisi dell\u2019espressione genica \ue8 ancora scarsamente predittiva mentre quella dei microRNA, regolatori post-trascrizionali dell\u2019espressione genica, e del proteoma pi\uf9 predittiva. Tali tecniche sono ancora costose e poco validate per la loro applicazione in Sanit\ue0 Pubblica. CONCLUSIONI: \uc8 verosimile che il rapido sviluppo di questi metodi porti alla loro applicazione nei sistemi sanitari europei, come gi\ue0 ad esempio avvenuto in alcuni paesi del nord Europa. Tuttavia lo sviluppo della medicina predittiva appare non giustificato e di difficile gestione se il rischio di patologia determinato da tali metodi non pu\uf2 essere modulato tramite interventi preventivi. Pertanto \ue8 necessario che il professionista della prevenzione, e cio\ue9 l\u2019Igienista, sia attivamente coinvolto in questi cambiamenti e nella loro applicazione alla Sanit\ue0 Pubblica affinch\ue9 la medicina predittiva si concretizzi in politiche di prevenzione. Infatti una mera indicazione dell\u2019esistenza di un rischio di patologia che non possa essere modulato sarebbe di assai dubbia utilit\ue0 per lo stato di salute della popolazione e dell\u2019individuo

Rationale for the use of N-acetylcysteine in both prevention and adjuvant therapy of COVID-19

COVID-19 may cause pneumonia, acute respiratory distress syndrome, cardiovascular alterations, and multiple organ failure, which have been ascribed to a cytokine storm, a systemic inflammatory response, and an attack by the immune system. Moreover, an oxidative stress imbalance has been demonstrated to occur in COVID-19 patients. N- Acetyl-L-cysteine (NAC) is a precursor of reduced glutathione (GSH). Due to its tolerability, this pleiotropic drug has been proposed not only as a mucolytic agent, but also as a preventive/therapeutic agent in a variety of disorders involving GSH depletion and oxidative stress. At very high doses, NAC is also used as an antidote against paracetamol intoxication. Thiols block the angiotensin-converting enzyme 2 thereby hampering penetration of SARS-CoV-2 into cells. Based on a broad range of antioxidant and anti-inflammatory mechanisms, which are herein reviewed, the oral administration of NAC is likely to attenuate the risk of developing COVID-19, as it was previously demonstrated for influenza and influenza-like illnesses. Moreover, high-dose intravenous NAC may be expected to play an adjuvant role in the treatment of severe COVID-19 cases and in the control of its lethal complications, also including pulmonary and cardiovascular adverse events

Alterazioni molecolari e morfologiche in spermatozoi di topi esposti al fumo di sigaretta

L'abitudine al fumo \ue8 considerata uno dei maggiori fattori di rischio per lo sviluppo del tumore al polmone e di un'ampia gamma di malattie cronico degenerative. Studi epidemiologici evidenziano chiaramente come il livello di fertilit\ue0 negli uomini sia in continuo decremento attribuendo il 13% dell'infertilit\ue0 al tabagismo. Il fumo di sigaretta (CS) \ue8 una miscela complessa contenente un gran numero di molecole cancerogene e mutagene. Alcune di queste molecole e/o i loro metaboliti possono attraversare la barriera ematotesticolare determinando effetti sulla fertilit\ue0. Lo studio \ue8 stato effettuato su un totale di 38 topi maschi albino Swiss CD1, di cui 13 costituivano il gruppo di controllo mentre i rimanenti 25 sono stati esposti fin dalla nascita al CS, ottenuto da sigarette di riferimento, per 10 settimane. Al termine di tale periodo gli animali sono stati sacrificati, sono stati prelevati gli spermatozoi dalla testa dell'epididimo per valutarne lo stato ossidativo, la funzionalit\ue0 mitocondriale, le alterazioni morfologiche, e la frammentazione del DNA spermatico. I risultati ottenuti confermano la capacit\ue0 del CS di indurre un aumento statisticamente significativo dello stress ossidativo (P<0,05), dei prodotti finali della perossidazione lipidica (P<0,05) e una maggiore frammentazione del DNA (P< 0,01). Inoltre l'analisi citologica effettuata su un numero pari a 1000 spermatozoi per singolo animale ha evidenziato una maggiore fragilit\ue0 spermatica (assenza della coda) nel gruppo degli animali esposti (P<0,05) ed alterazioni morfologiche quali assenza del gancio (P<0,05), testa a spillo (P<0,05) e spermatozoi con due code (P<0,05). I dati ottenuti evidenziano come l'abitudine al fumo possa avere un rimarchevole effetto sulla fertilit\ue0 maschile. Questo se da una parte \ue8 imputabile all'induzione dello stress ossidativo, come evidenziato dall'aumento delle specie reattive dell'ossigeno e dai prodotti della perossidazione lipidica, dall'altra pu\uf2 essere determinato dall'effetto mutageno esercitato dalle componenti aromatiche del CS, come dimostrato dall'analisi morfologica e dalla frammentazione del DNA

Asbestiform amphiboles and cleavage fragments analogues: Overview of critical dimensions, aspect ratios, exposure and health effects

The term asbestos refers to a group of serpentine (chrysotile) and amphibole (amosite, crocidolite, anthophyllite, tremolite and actinolite) minerals with a fibrous habit. Their chemical-physical properties make them one of the most important inorganic materials for industrial purposes and technological applications. However, the extraction, use and marketing of these minerals have been prohibited due to proven harmful effects, mainly involving the respiratory system. In addition to the known six minerals classified as asbestos, the natural amphiboles and serpentine polymorphs antigorite and lizardite, despite having the same composition of asbestos, do not have the same morphology. These minerals develop chemical and geometric (length &gt; 5 \ub5m, width &lt; 3 \ub5m and length: diameter &gt; 3:1), but not morphological, analogies with asbestos, which is regulated by the WHO. The debate about their potential hazardous properties is open and ongoing; therefore, their morphological characterization has a key role in establishing a reliable asbestos hazard scenario. This review focuses on evaluating the most relevant papers, evidencing the need for a reappraisal. Different in vitro, in vivo and epidemiological studies report information about cleavage fragments with critical dimensions similar to asbestos fibres, but very few works target fragments below 5 \ub5m in length. Breathable smaller fibres could have deleterious effects on human health and cannot be disregarded from the risk assessment process. Furthermore, a few studies suggest that the carcinogenic nature of short fibres is not excluded. This review highlights that it is worth investigating the effects of this size range of elongated mineral particles and fibres