Ultra-wide-field fundus photography compared to ophthalmoscopy in diagnosing and classifying major retinal diseases.

To analyze the performance of ultra-wide-field (UWF) fundus photography compared with ophthalmoscopy in identifying and classifying retinal diseases. Patients examined for presumed major retinal disorders were consecutively enrolled. Each patient underwent indirect ophthalmoscopic evaluation, with scleral depression and/or fundus biomicroscopy, when clinically indicated, and mydriatic UWF fundus imaging by means of CLARUS 500™ fundus camera. Each eye was classified by a clinical grader and two image graders in the following groups: normal retina, diabetic retinopathy, vascular abnormalities, macular degenerations and dystrophies, retinal and choroidal tumors, peripheral degenerative lesions and retinal detachment and myopic alterations. 7024 eyes of new patients were included. The inter-grader agreement for images classification was perfect (kappa = 0.998, 95% Confidence Interval (95%CI) = 0.997-0.999), as the two methods concordance for retinal diseases diagnosis (kappa = 0.997, 95%CI = 0.996-0.999) without statistically significant difference. UWF fundus imaging might be an alternative to ophthalmoscopy, since it allows to accurately classify major retinal diseases, widening the range of disorders possibly diagnosed with teleophthalmology. Although the clinician should be aware of the possibility that a minority of the most peripheral lesions may be not entirely visualized, it might be considered a first line diagnostic modality, in the context of a full ophthalmological examination.

The miniaturized enzyme-modified comet assay for genotoxicity testing of nanomaterials.

The in vitro comet assay is a widely applied method for investigating genotoxicity of chemicals including engineered nanomaterials (NMs). A big challenge in hazard assessment of NMs is possible interference between the NMs and reagents or read-out of the test assay, leading to a risk of biased results. Here, we describe both the standard alkaline version of the in vitro comet assay with 12 mini-gels per slide for detection of DNA strand breaks and the enzyme-modified version that allows detection of oxidized DNA bases by applying lesion-specific endonucleases (e.g., formamidopyrimidine DNA glycosylase or endonuclease III). We highlight critical points that need to be taken into consideration when assessing the genotoxicity of NMs, as well as basic methodological considerations, such as the importance of carrying out physicochemical characterization of the NMs and investigating uptake and cytotoxicity. Also, experimental design-including treatment conditions, cell number, cell culture, format and volume of medium on the plate-is crucial and can have an impact on the results, especially when testing NMs. Toxicity of NMs depends upon physicochemical properties that change depending on the environment. To facilitate testing of numerous NMs with distinct modifications, the higher throughput miniaturized version of the comet assay is essential.

Particle size, chemical composition, seasons of the year and urban, rural or remote site origins as determinants of biological effects of particulate matter on pulmonary cells.

Particulate matter (PM), a complex mix of chemical compounds, results to be associated with various health effects. However there is still lack of information on the impact of its different components. PM2.5 and PM1 samples, collected during the different seasons at an urban, rural and remote site, were chemically characterized and the biological effects induced on A549 cells were assessed. A Partial Least Square Discriminant Analysis has been performed to relate PM chemical composition to the toxic effects observed. Results show that PM-induced biological effects changed with the seasons and sites, and such variations may be explained by chemical constituents of PM, derived both from primary and secondary sources. The first-time here reported biological responses induced by PM from a remote site at high altitude were associated with the high concentrations of metals and secondary species typical of the free tropospheric aerosol, influenced by long range transports and aging.

Season linked responses to fine and quasi-ultrafine Milan PM in cultured cells.

Exposure to urbane airborne particulate matter (PM) is related to the onset and exacerbation of cardiovascular and respiratory diseases. The fine (PM1), and quasi-ultrafine (PM0.4) Milan particles collected during different seasons have been characterised and the biological effects on human epithelial lung A549, monocytes THP-1 cells and their co-culture, evaluated and compared with the results obtained on the PM10 and PM2.5 fractions. Chemical composition and transmission electron microscopy (TEM) analysis of PM0.4 showed that this fraction was very similar to PM1 for biological responses and dimension. All the winter fractions increased within 1h the level of reactive oxygen species (ROS), while only summer PM2.5 had this effect on A549 cells. The phosphorylation of H2AX (γH2AX), a marker of double strand DNA breaks (DSBs), was increased by all the winter fractions on A549 and THP-1 cells while summer PM samples did not induced this effect. PM0.4 and PM1 biological effects are partly similar and related to the season of sampling, with effects on ROS and DNA damage induced only by winter PM fractions. The winter PM damaging effect on DNA correlates with the presence of organic compounds.