Outdoor airborne allergens: Characterization, behavior and monitoring in Europe.

Aeroallergens or inhalant allergens, are proteins dispersed through the air and have the potential to induce allergic conditions such as rhinitis, conjunctivitis, and asthma. Outdoor aeroallergens are found predominantly in pollen grains and fungal spores, which are allergen carriers. Aeroallergens from pollen and fungi have seasonal emission patterns that correlate with plant pollination and fungal sporulation and are strongly associated with atmospheric weather conditions. They are released when allergen carriers come in contact with the respiratory system, e.g. the nasal mucosa. In addition, due to the rupture of allergen carriers, airborne allergen molecules may be released directly into the air in the form of micronic and submicronic particles (cytoplasmic debris, cell wall fragments, droplets etc.) or adhered onto other airborne particulate matter. Therefore, aeroallergen detection strategies must consider, in addition to the allergen carriers, the allergen molecules themselves. This review article aims to present the current knowledge on inhalant allergens in the outdoor environment, their structure, localization, and factors affecting their production, transformation, release or degradation. In addition, methods for collecting and quantifying aeroallergens are listed and thoroughly discussed. Finally, the knowledge gaps, challenges and implications associated with aeroallergen analysis are described.

Integrative toolbox to assess the quality of freshwater sediments contaminated with potentially toxic metals.

The Guadiana Basin is a transnational basin, presenting historical contamination with potentially toxic metals (PTM), which origin can be both natural and anthropogenic. This study explores the use of a set of observational, chemical and ecotoxicological assays with Heterocypris incongruens, Vibrio fischeri, Pseudokirchneriella subcapitata, Thamnocephalus platyurus, identifying the most sensitive to be included in a toolbox to analyze the quality of freshwater sediments related to this type of contamination. The study included the analysis of a reservoir and streams sediments of Guadiana basin, in two consecutive years with different climate conditions 2017 (dry year) and 2018 (normal year). The results showed high chemical variability along the basin, with greater contamination with PTM in the reservoir sediments. The calculated Enrichment Factors (EF) indicated high anthropogenic contamination by Cd, followed by Pb (EF > 1.5). The geoaccumulation index (Igeo) revealed that the sediments were severely polluted with Cd, and slightly polluted with Pb and Cu, inducing a higher sublethal toxicity to Heterocypris incongruens. Among the parameters evaluated, and after the use of multivariate statistical techniques, the toolbox for assessing sediments quality, in similar climate and geological conditions, should include the analysis of: meteorology, land use/cover in the area, granulometry, organic matter content, PTM concentrations, contamination indices (e.g., Igeo and EF), and sublethal bioassays with H. incongruens (total sediment analysis) and Vibrio fisheri luminescence inhibition (pore water analysis).

Benthic diatom community dynamics in Mediterranean intermittent streams: Effects of water availability and their potential as indicators of dry-phase ecological status.

The study of intermittent rivers is a critical and timely issue due to their worldwide increase, triggered by several causes including climate change. The need to understand the response of intermittent river biota to water intermittency led us to conduct this study using benthic diatoms collected in southern Portugal. Benthic diatoms were explored in terms of assemblages, diversity indices, the Specific Pollution Sensitivity (SPI) Index, functional metrics (i.e. ecological guilds and life-forms) and conservation status. We verified that changes in water physico-chemical characteristics were highly controlled by flow intermittency, which in turn is directly linked to meteorological variables (air temperature and precipitation). Changes in diatom assemblages reflect the aquatic regime of sites, changes in aquatic states through time and mesohabitats (dry biofilm, samples collected in pools or under flowing conditions). Species richness, on the other hand, did not reflect these differences, whilst Shannon diversity and Pielou's Evenness indices only reflected mesohabitat differences. The SPI distinguished sampling periods, and mesohabitats. The relative abundance of ecological guilds changed with aquatic states, with the low-profile guild dominating in eurheic and arheic conditions (except during Summer), being replaced by motile taxa in summer arheic conditions, reflecting increases in nutrient and siltation. The hypothesis that benthic diatom assemblages in dry biofilm can be used as an indicator of ecological status during the dry-phase was validated, since no differences between the Ecological Quality Ratio determined in dry biofilm collected in Summer 2017 and the previous Spring 2017 in flowing water. A method is proposed for diatom sampling in dry biofilm, contributing to an integrated ecological status evaluation, which considers the dry-phase and enhances the reach of biomonitoring programs.

Vertical distribution of benthic diatoms in a large reservoir (Alqueva, Southern Portugal) during thermal stratification.

Freshwater diatom communities are known to respond to a wide range of environmental factors, however, the depth gradient is usually neglected and few studies are available, especially in large reservoirs. During the ALqueva hydro-meteorological EXperiment (ALEX) field campaign, diatom communities were studied in the margins and in three platforms (from the surface to the bottom of the reservoir) located in the limnetic zone of the Alqueva reservoir, one of the largest artificial lakes in western Europe. A detailed meteorological and physico-chemical characterization of the reservoir was carried out from June to September in Summer 2014, when the reservoir was stratified, to relate these variables with diatom assemblages. Despite the large dimensions of the reservoir, no differences in the water physico-chemical characteristics and diatom descriptors were detected among platforms. Small changes in diatom assemblages, ecological guilds, taxa richness and Shannon diversity index were observed between sampling campaigns. Nevertheless, differences in diatoms were detected along a depth gradient, both in terms of diatom assemblages and ecological guilds. Taxa richness, Shannon diversity index, Pielou's evenness and Specific Pollution sensitivity Index (SPI) also differed with depth, with the lowest values of all indices detected at surface samples, increasing with depth, reaching the highest values at 20 m for taxa richness, Shannon diversity and Pielou's evenness indices.

Effects of intravitreal insulin and insulin signaling cascade inhibitors on emmetropization in the chick.

PURPOSE: Intravitreal insulin has been shown to be a powerful stimulator of myopia in chickens, in particular if the retinal image is degraded or defocused. In most tissues, the insulin receptor activates two main signaling pathways: a) the mitogen-activated protein kinase (MAPK) cascade (e.g., mitogen-activated protein kinasem kinase [MEK] and extracellular regulated kinase [ERK]) and b) the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway. In the current study, insulin was injected, and these pathways were separately inhibited to determine which is activated when the retinal image is defocused by spectacle lenses. METHODS: Chicks were treated with either +7 D, -7 D, or no lenses. They were intravitreally injected with insulin, the MEK inhibitor U0126, the PI3K inhibitor Ly294002, or a combination of insulin and one of the inhibitors. Refractions and ocular dimension were measured at the beginning and after four days of treatment. The retinal proteins of the chicks were measured with western blots after 2 h and four days of treatment. Incubation occurred with anti-Akt1, anti-Erk1/2, anti-phospho-Akt(Thr308), and anti-phospho-Erk1/2((Thr202/Tyr204)) antibodies, and the ratio between the relative intensity of the phospho-form and the total-form was calculated. RESULTS: Chicks wearing positive lenses and injected with saline and with PI3K inhibitor compensated for the imposed defocus and became hyperopic. Insulin injections and insulin plus PI3K inhibitor injections prevented lens-induced hyperopia, whereas the MEK inhibitor alone and insulin plus MEK inhibitor had no effect. Obviously, the MEK inhibitor suppressed the effect of insulin on eye growth in the plus lens-treated animals. Chicks treated with negative lenses and injected with insulin, or with insulin plus MEK inhibitor, overcompensated for the imposed defocus. This effect of insulin was not detected in eyes injected with PI3K inhibitor plus insulin, suggesting that the PI3K inhibitor suppressed the effects of insulin in minus lens-treated animals. Insulin increased the ratio of phospho-Akt/total-Akt in animals with normal visual exposure but even more so in chicks wearing plus or minus lenses. The increase was blocked by simultaneous PI3K inhibitor injections in control eyes but not in lens-treated eyes. Insulin also increased the ratio of phospho-ERK/total-ERK in animals with normal visual exposure and in animals wearing positive lenses, compared to U0126- and Ly294002-injected eyes. In contrast, no significant activation of the MEK/ERK pathway was observed in the negative lens-treated animals. CONCLUSIONS: Intravitreal insulin promoted axial eye growth and stimulated both signaling pathways. The PI3K/Akt pathway was activated in control and plus and minus lens-treated eyes, but the MEK/ERK pathway was activated only with positive lenses or no lenses. With negative lenses, insulin did not stimulate the MEK/ERK signaling cascade. Independent of the pathway stimulated after insulin binding, the effect on insulin was always the same: an increase in eye growth.

Ultrasonography and optical low-coherence interferometry compared in the chicken eye.

PURPOSE: To compare ocular biometry [anterior chamber depth (ACD), lens thickness (LT), vitreous chamber depth (VCD), and axial length (AL)] using A-scan ultrasonography and optical low-coherence interferometry (OLCI) in the chicken eye. METHODS: Two-week-old chicks (n = 42) were measured. Bland-Altman plots and repeatability and correlation analyses were calculated for both methods. RESULTS: There was a high correlation between both methods for ACD (r = 0.6144, p < 0.0001), VCD (r = 0.9595, p < 0.0001), and AL (r = 0.9290, p < 0.0001) but not for LT (r = 0.1604, p = 0.144). Measurements by OLCI were more consistent (smaller coefficients of variation and higher intraclass correlation). Bland-Altman plots showed that ultrasound provided larger values for LT, VCD, and AL but not for ACD [differences between ultrasound and OLCI (mean ± SD): ACD = -0.11 ± 0.12 mm; LT = 0.10 ± 0.09 mm; VCD = 0.25 ± 0.08 mm; AL = 0.50 ± 0.16 mm]. CONCLUSIONS: A high correlation between both techniques was found for three of the four parameters (ACD, VCD, and AL). However, as the absolute values were different, both techniques cannot replace each other mainly because (1) one is non-contact and the other contact and can induce a minor indentation of the cornea and (2) each device uses different types of waves that cross the ocular interfaces differently. While consistency and repeatability were better by OLCI, a disadvantage is that, different from humans, it can only be used in anesthetized chicks.

Insulin, insulin-like growth factor-1, insulin receptor, and insulin-like growth factor-1 receptor expression in the chick eye and their regulation with imposed myopic or hyperopic defocus.

PURPOSE: Insulin stimulates eye growth in chicks and this effect is greatly enhanced if the retinal image is degraded by the defocus of either sign. However, it is unclear whether the insulin receptor (IR) is expressed at all in the chicken retina in animals 1-2 weeks post-hatching. We have investigated IR expression and whether IR transcript abundance varies in the fundal layers. To elucidate the possible role of insulin and insulin-like growth factor (IGF)-1 signaling in eye growth regulation, mRNA (mRNA) levels were measured for insulin, IGF-1, IR, and IGF-1 receptor (IGF-1R) during imposed negative or positive defocus. METHODS: Chicks were treated binocularly with positive or negative spectacle lenses for 4 or 24 h, or they remained untreated (n=6, for each treatment group). Northern blot analyses were performed to screen for transcription variants in the different fundal layers of untreated animals. Real-time PCR was used to quantify IR, IGF-1R, IGF-1, and insulin mRNA levels in the different fundal layers of the chick eye in the three treatment groups. RESULTS: IR mRNA was found in all the studied tissues, although there is evidence of tissue-specific transcript variations. Three major transcripts were detected for IR. The brain, retina, and choroid showed the longest transcript (4.3 kb), which was not present in the liver. Nevertheless, the liver and brain showed a second transcript (2.6 kb) not present in the retina and choroid. A short transcript (1.3 kb) was the predominant form in the liver and choroid, and it seems to be present in the retinal pigment epithelium (RPE) and sclera as well. In the retina, no significant gene expression changes were found when defocus was imposed. Interestingly, in the RPE, both IR and IGF-1R were already downregulated after short periods (4 h) of positive lens wear. In contrast, IR and IGF-1R were upregulated in the choroid and fibrous sclera during treatment with negative, but not positive, lenses. CONCLUSIONS: Differences observed in the IR transcript length in different tissues suggest possibly different functions. The differential regulation of IR and IGF-1R in the RPE, choroid, and fibrous sclera is consistent with their involvement in a signaling cascade for emmetropization.