A 4-year study of bovine reproductive hormones that are induced by pharmaceuticals and appear as steroid estrogenic pollutants in the resulting slurry, using in vitro and instrumental analytical methods.

The main objective of the research was to study the environmental "price" of the large-scale, milk production from a rarely known perspective, from the mapping of the estrogenic footprint (the amount of oestrus-inducer hormonal products, and the generated endoestrogens) in the resulting slurry in a dairy cow farm. These micropollutants are endocrine-disrupting chemicals (EDCs) and can be dangerous to the normal reproductive functions even at ng/kg concentration. One of them, 17ß-estradiol, has a 20,000 times stronger estrogenic effect than bisphenol-A, a widely known EDC of industrial origin. While most studies on EDCs are short-term and/or laboratory based, this study is longitudinal and field-based. We sampled the slurry pool on a quarterly basis between 2017 and 2020. Our purpose was testing the estrogenic effects using a dual approach. As an effect-based, holistic method, we developed and used the YES (yeast estrogen screen) test employing the genetically modified Saccharomyces cerevisiae BJ3505 strain which contains human estrogenic receptor. For testing exact molecules, UHPLC-FLD was used. Our study points out that slurry contains a growing amount of EDCs with the risk of penetrating into the soil, crops and the food chain. Considering the Green Chemistry concept, the most benign ways to prevent of the pollution of the slurry is choosing appropriate oestrus-inducing veterinary pharmaceuticals (OIVPs) and the separation of the solid and liquid parts with adequate treatment methods. To our knowledge, this is the first paper on the adaptation of the YES test for medicine and slurry samples, extending its applicability. The adapted YES test turned out to be a sensitive, robust and reliable method for testing samples with potential estrogenic effect. Our dual approach was successful in evaluating the estrogenic effect of the slurry samples.

Effect of prevailing winds and land use on Alternaria airborne spore load.

Alternaria spores are a common component of the bioaerosol. Many Alternaria species are plant pathogens, and their conidia are catalogued as important aeroallergens. Several aerobiological studies showing a strong relationship between concentrations of airborne spore and meteorological parameters have consequently been developed. However, the Alternaria airborne load variation has not been thoroughly investigated because it is difficult to assess their sources, as they are a very common and widely established phytopathogen. The objective of this study is to estimate the impact of vegetation and land uses as potential sources on airborne spore load and to know their influence, particularly, in cases of long-medium distance transport. The daily airborne spore concentration was studied over a 5-year period in León and Valladolid, two localities of Castilla y León (Spain), with differences in their bioclimatic and land use aspects. Moreover, the land use analysis carried out within a 30 km radius of each monitoring station was combined with air mass data in order to search for potential emission sources. The results showed a great spatial variation between the two areas, which are relatively close to each other. The fact that the spore concentrations recorded in Valladolid were higher than those in León was owing to prevailing winds originating from large areas covered by cereal crops, especially during the harvest period. However, the prevailing winds in León came from areas dominated by forest and shrubland, which explains the low airborne spore load, since the main Alternaria sources were the grasslands located next to the trap. Furthermore, the risk days in this location presented an unusual wind direction. This study reveals the importance of land cover and wind speed and direction data for establishing potential airborne routes of spore transport in order to improve the Alternaria forecasting models. The importance of conducting Alternaria aerobiological studies at a local level is also highlighted.

Turning Red without Feeling Embarrassed─Xanthenium-Based Photocages for Red-Light-Activated Phototherapeutics.

Herein, we present high-yielding, concise access to a set of xanthenium-derived, water-soluble, low-molecular-weight photocages allowing light-controlled cargo release in the green to red region. Very importantly, these new photocages allow installation of various payloads through ester, carbamate, or carbonate linkages even at the last stage of the synthesis. Payloads were uncaged with high efficiency upon green, orange, or red light irradiation, leading to the release of carboxylic acids, phenols, and amines. The near-ideal properties of a carboxanthenium derivative were further evaluated in the context of light-controlled drug release using a camptothecin-derived chemotherapeutic drug, SN38. Notably, the caged drug showed orders of magnitude lower efficiency in cellulo, which was reinstated after red light irradiation. The presented photocages offer properties that facilitate the translation of photoactivated chemotherapy toward clinical applications.

A Genetically Encoded Isonitrile Lysine for Orthogonal Bioorthogonal Labeling Schemes.

Bioorthogonal click-reactions represent ideal means for labeling biomolecules selectively and specifically with suitable small synthetic dyes. Genetic code expansion (GCE) technology enables efficient site-selective installation of bioorthogonal handles onto proteins of interest (POIs). Incorporation of bioorthogonalized non-canonical amino acids is a minimally perturbing means of enabling the study of proteins in their native environment. The growing demand for the multiple modification of POIs has triggered the quest for developing orthogonal bioorthogonal reactions that allow simultaneous modification of biomolecules. The recently reported bioorthogonal [4 + 1] cycloaddition reaction of bulky tetrazines and sterically demanding isonitriles has prompted us to develop a non-canonical amino acid (ncAA) bearing a suitable isonitrile function. Herein we disclose the synthesis and genetic incorporation of this ncAA together with studies aiming at assessing the mutual orthogonality between its reaction with bulky tetrazines and the inverse electron demand Diels-Alder (IEDDA) reaction of bicyclononyne (BCN) and tetrazine. Results showed that the new ncAA, bulky-isonitrile-carbamate-lysine (BICK) is efficiently and specifically incorporated into proteins by genetic code expansion, and despite the slow [4 + 1] cycloaddition, enables the labeling of outer membrane receptors such as insulin receptor (IR) with a membrane-impermeable dye. Furthermore, double labeling of protein structures in live and fixed mammalian cells was achieved using the mutually orthogonal bioorthogonal IEDDA and [4 + 1] cycloaddition reaction pair, by introducing BICK through GCE and BCN through a HaloTag technique.

Towards a model of wet deposition of bioaerosols: The raindrop size role.

Bioaerosols play a major role in the plant life of ecosystems. In addition, they have a profound impact on human health, since they may cause lung diseases or allergies. The key objective of this study is to assess the below cloud scavenging effect of rainfall on pollen concentration. The analysis is based on a sampling carried out in León, Spain, between 2015 and 2018. The rainfall variables and the pollen concentrations have been obtained with a disdrometer and a volumetric Hirst type spore-trap, respectively. In order to evaluate the scavenging, three parameters have been calculated: scavenging efficiency (through the concentration-weighted average (%ΔC)), the scavenging coefficient (λ) and the percentage of events with a decrease in pollen concentration (%ES) also called events with effective scavenging. 71% of rain events presented an effective scavenging that affected all types of pollen. The %ΔC mean value of total pollen was 24 ± 18% (positive values indicate an effective scavenging) and the types of pollen with the highest values were Castanea and Cupressaceae (71 and 40%, respectively). A linear model (R2 = 0.94) to estimate the pollen concentration after rain was built with variables such as pollen concentration before rain and other variables from a weather station and a disdrometer. Furthermore, we have shown the possibility of knowing in real time the probable Cupressaceae pollen concentration, from the initial pollen concentration and the physical parameters of rain (such as raindrop size, rain intensity or volume swept by raindrops in their falling path).

West Nile virus 'circulation' in Vojvodina, Serbia: Mosquito, bird, horse and human surveillance.

Efforts to detect West Nile virus (WNV) in the Vojvodina province, northern Serbia, commenced with human and mosquito surveillance in 2005, followed by horse (2009) and wild bird (2012) surveillance. The knowledge obtained regarding WNV circulation, combined with the need for timely detection of virus activity and risk assessment resulted in the implementation of a national surveillance programme integrating mosquito, horse and bird surveillance in 2014. From 2013, the system showed highly satisfactory results in terms of area specificity (the capacity to indicate the spatial distribution of the risk for human cases of West Nile neuroinvasive disease - WNND) and sensitivity to detect virus circulation even at the enzootic level. A small number (n = 50) of Culex pipiens (pipiens and molestus biotypes, and their hybrids) females analysed per trap/night, combined with a high number of specimens in the sample, provided variable results in the early detection capacity at different administrative levels (NUTS2 versus NUTS3). The clustering of infected mosquitoes, horses, birds and human cases of WNND in 2014-2015 was highly significant, following the south-west to north-east direction in Vojvodina (NUTS2 administrative level). Human WNND cases grouped closest with infected mosquitoes in 2014, and with wild birds/mosquitoes in 2015. In 2014, sentinel horses showed better spatial correspondence with human WNND cases than sentinel chickens. Strong correlations were observed between the vector index values and the incidence of human WNND cases recorded at the NUTS2 and NUTS3 levels. From 2010, West Nile virus was detected in mosquitoes sampled at 43 different trap stations across Vojvodina. At 14 stations (32.56%), WNV was detected in two different (consecutive or alternate) years, at 2 stations in 3 different years, and in 1 station during 5 different years. Based on these results, integrated surveillance will be progressively improved to allow evidence-based adoption of preventive public health and mosquito control measures.