Microscopic computed tomography with AI-CNN-powered image analysis: the path to phenotype bleomycin-induced lung injury.

Bleomycin (BLM)-induced lung injury in mice is a valuable model for investigating the molecular mechanisms that drive inflammation and fibrosis and for evaluating potential therapeutic approaches to treat the disease. Given high variability in the BLM model, it is critical to accurately phenotype the animals in the course of an experiment. In the present study, we aimed to demonstrate the utility of microscopic computed tomography (µCT) imaging combined with an artificial intelligence (AI)-convolutional neural network (CNN)-powered lung segmentation for rapid phenotyping of BLM mice. µCT was performed in freely breathing C57BL/6J mice under isoflurane anesthesia on days 7 and 21 after BLM administration. Terminal invasive lung function measurement and histological assessment of the left lung collagen content were conducted as well. µCT image analysis demonstrated gradual and time-dependent development of lung injury as evident by alterations in the lung density, air-to-tissue volume ratio, and lung aeration in mice treated with BLM. The right and left lung were unequally affected. µCT-derived parameters such as lung density, air-to-tissue volume ratio, and nonaerated lung volume correlated well with the invasive lung function measurement and left lung collagen content. Our study demonstrates the utility of AI-CNN-powered µCT image analysis for rapid and accurate phenotyping of BLM mice in the course of disease development and progression.NEW & NOTEWORTHY Microscopic computed tomography (µCT) imaging combined with an artificial intelligence (AI)-convolutional neural network (CNN)-powered lung segmentation is a rapid and powerful tool for noninvasive phenotyping of bleomycin mice over the course of the disease. This, in turn, allows earlier and more reliable identification of therapeutic effects of new drug candidates, ultimately leading to the reduction of unnecessary procedures in animals in pharmacological research.

Circulating Microparticles Are Differentially Increased in Lowlanders and Highlanders with High Altitude Induced Pulmonary Hypertension during the Cold Season.

The role of microparticles (MPs) and cold in high altitude pulmonary hypertension (HAPH) remains unexplored. We investigated the impact of long-term cold exposure on the pulmonary circulation in lowlanders and high-altitude natives and the role of MPs. Pulmonary hemodynamics were evaluated using Doppler echocardiography at the end of the colder and warmer seasons. We further examined the miRNA content of MPs isolated from the study participants and studied their effects on human pulmonary artery smooth muscle (hPASMCs) and endothelial cells (hPAECs). Long-term exposure to cold environment was associated with an enhanced pulmonary artery pressure in highlanders. Plasma levels of CD62E-positive and CD68-positive MPs increased in response to cold in lowlanders and HAPH highlanders. The miRNA-210 expression contained in MPs differentially changed in response to cold in lowlanders and highlanders. MPs isolated from lowlanders and highlanders increased proliferation and reduced apoptosis of hPASMCs. Further, MPs isolated from warm-exposed HAPH highlanders and cold-exposed highlanders exerted the most pronounced effects on VEGF expression in hPAECs. We demonstrated that prolonged exposure to cold is associated with elevated pulmonary artery pressures, which are most pronounced in high-altitude residents. Further, the numbers of circulating MPs are differentially increased in lowlanders and HAPH highlanders during the colder season.

Diversity of Beauveria spp. isolates from pollen beetles Meligethes aeneus in Switzerland.

Pollen beetles Meligethes aeneus were collected in oilseed rape fields at different sites in Switzerland in spring 2004-2005 and 32 isolates of the fungal genus Beauveria occurring as latent infections in the beetles were obtained and molecularly characterized. Three major clades, Beauveria bassiana sensu stricto (Clade A: n=13), Beauveriabrongniartii (Clade B: n=1) and Beauveria Clade C (n=18) were identified among the isolates based on sequences of the ITS region and the 5' end of EF1-α. B. bassiana s.s. was further separated in the two clades, Eu_1 (n=10) and Eu_4 (n=3). The intergenic region Bloc provided best resolution of the individual clades B. bassiana s.s. Eu_1, Eu_4 and B. brongniartii. No specific clade of Beauveria appeared to be associated with adult M. aeneus populations. However, data suggested high relative abundance of Beauveria Clade C among the fungal entomopathogens infecting M. aeneus. Characterization of the isolates by simple sequence repeats (SSR) revealed further genotypic diversity within the clades except B. bassiana s.s. Eu_4 which appeared to be clonal. However, the individual SSR markers were differentially amplifiable from isolates of the different clades. It is therefore important to identify the underlying phylogenetic affinity of Beauveria isolates to interpret results based on SSR markers. The data suggest that not all available SSR markers are suitable for reliable characterization of diversity within Beauveria Clade C.

Susceptibility of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) to the entomopathogenic fungus Metarhizium anisopliae when feeding on Bacillus thuringiensis Cry3Bb1-expressing maize.

Genetically engineered maize producing the insecticidal protein Cry3Bb1 from Bacillus thuringiensis (Bt maize) is protected against corn rootworms (Diabrotica spp.), which are serious maize pests in North America and Europe. The aim of the present study was to investigate the interaction of Bt maize (event MON88017) and the entomopathogenic fungus Metarhizium anisopliae for controlling the western corn rootworm, Diabrotica virgifera virgifera. Exposure to Cry3Bb1 expressed in Bt maize seedlings resulted in decreased weight gain in D. v. virgifera larvae but did not influence susceptibility to M. anisopliae. Adult beetles were not affected by Cry3Bb1 in their food, but mortality when feeding on maize leaves was higher than when feeding on silk. Adults were more susceptible to the fungus than larvae. The results indicate that the effects of Bt maize and M. anisopliae on D. v. virgifera are additive and that Bt maize does not interfere with the biological control provided by entomopathogenic fungi.