High-throughput mapping of a dynamic signaling network in mammalian cells

Signaling pathways transmit information through protein interaction networks that are dynamically regulated by complex extracellular cues. We developed LUMIER (for luminescence-based mammalian interactome mapping), an automated high-throughput technology, to map protein-protein interaction networks systematically in mammalian cells and applied it to the transforming growth factor-beta (TGFbeta) pathway. Analysis using self-organizing maps and k-means clustering identified links of the TGFbeta pathway to the p21-activated kinase (PAK) network, to the polarity complex, and to Occludin, a structural component of tight junctions. We show that Occludin regulates TGFbeta type I receptor localization for efficient TGFbeta-dependent dissolution of tight junctions during epithelial-to-mesenchymal transitions.     

Investigations of the maintenance system of the Konik Polski horse and its effects on fecal microbiota activity during the winter and summer seasons

Gastrointestinal microbiota play a key role in the nutrients digestion and hence maintaining animal health and welfare. The diet offered to the animals in captivity may differ considerably from that on natural pastures. In a stabled maintenance system, horses have a limited choice of habitat and feed. Time spend for feeding is relevant for equine welfare because the reduction of the time devoted for foraging may be responsible for inducing gastric inflammation and ulceration. Therefore, in the present study, it was hypothesized that fecal bacterial fermentative processes differ between free-roaming and stabled Konik Polski Horses (KPHs) with respect to microbial enzymatic activity, and thus the fecal concentration of short-chain fatty acids (SCFA), which can be further utilized for assessing the feeding behavior and welfare in free-roaming versus stabled horses. The SCFA concentration and profile, as well as the extracellular and intracellular activities of selected bacterial enzymes, were characterized in horse feces collected during the winter and summer feeding seasons. The results showed higher enzymatic activity and SCFA production in the feces excreted by free-roaming versus stabled horses, especially during summer. An increase in pasture plant diversity may be beneficial for the gastrointestinal microbiota and hence for maintaining health and welfare.     

Biomarkers of oxidative stress and antioxidant defences as indicators of different disinfectants exposure in the heart of rainbow trout (Oncorhynchus Mykiss Walbaum)

The aim of this study was to evaluate the impacts of different disinfectants' treatment using in aquaculture on the oxidative stress biomarkers such as thiobarbituric acid reactive substrates (TBARS) and carbonyl derivatives of protein oxidative modification, as well as antioxidant defences [superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx) activity] and total antioxidant capacity (TAC) in the heart tissue of rainbow trout (Oncorhynchus mykiss). In the disinfectants exposure, fish were treated to chloramine‐T (final concentration 10 g m^?3), chloride dioxide (5 g m^?3), formalin (200 mL m^?3) and CIP disinfectant based on peracetic acid and hydrogen peroxide (16 mL m^?3) for 20 min and repeated three times every 3 days. Both chlorine dioxide and formalin treatment was indicated by a significant increase in the level of heart TBARS levels and carbonyl derivatives content and decreased SOD activity. Tissue oxidative stress biomarkers were unchanged upon chloramine‐T or CIP disinfectant exposure. Increased oxidative stress could modify antioxidant defences, principally causing increased CAT activity in the heart tissue of formalin‐ or ClO₂^?‐exposed fish. The correlation between oxidative stress biomarkers and GPx activity indicates that enzymes related to glutathione metabolism were responsible to formalin or ClO₂^?‐induced oxidative stress. Hence, TBARS, carbonyl derivatives and antioxidant defences could be used as biomarkers in evaluating the toxicity of formalin and chlorine dioxide using as disinfectants to trout.     

Feline hip joint anatomy in magnetic resonance images

The aim of this study was to develop an anatomical model of the feline hip joint for low‐field magnetic resonance imaging (LF‐MRI) based on high‐field magnetic resonance imaging (HF‐MRI). The study was performed on six adult clinically healthy European shorthair cats, aged 1–3 years, with body weight of 2.8–4.4 kg. The animals were examined with the use of the Vet‐MRI Grande Esaote LF (0.25 T) scanner and high‐field Siemens Magnetom TRIO (3 T) MRI scanner. In the LF‐MRI, most satisfactory results in T1‐weighted images were obtained when TE was 26 ms in all three planes and when TR was 350–950 ms in the transverse plane, 950–1150 ms in the sagittal plane and 520–750 ms in the dorsal plane. In T2‐weighted images, TE was 90 ms in the transverse and dorsal plane and 120 ms in the sagittal plane. The results were presented as images acquired with LF‐MRI scanners in three planes. The slice thickness was 3 mm for each plane. In LF‐MRI, muscles in the hip joint region and round ligament were well visualized. Unlike in LF‐MRI, the cross section of the femoral nerve was identified in HF‐MRI scans. In examinations of the feline hip joint, the main limitations of LF‐MRI were a lack of reliable contrast between articular cartilage and synovial fluid as well as longer scan time. Despite the above, LF‐MRI images were characterized by good contrast between bones and the surrounding soft tissues.     

Isolating the effect of soil properties on agricultural soil greenhouse gas emissions under controlled conditions

Agricultural soils are important sources of greenhouse gases (GHGs). Soil properties and environmental factors have complex interactions which influence the dynamics of these GHG fluxes. Four arable and five grassland soils which represent the range of soil textures and climatic conditions of the main agricultural areas in the UK were incubated at two different moisture contents (50 or 80% water holding capacity) and with or without inorganic fertiliser application (70 kg N ha⁻¹ ammonium nitrate) over 22 days. Emissions of N₂O, CO₂ and CH₄ were measured twice per week by headspace gas sampling, and cumulative fluxes were calculated. Multiple regression modelling was carried out to determine which factors (soil mineral N, organic carbon and total nitrogen contents, C:N ratios, clay contents and pH) that best explained the variation in GHG fluxes. Clay, mineral N and soil C contents were found to be the most important explanatory variables controlling GHG fluxes in this study. However, none of the measured variables explained a significant amount of variation in CO₂ fluxes from the arable soils. The results were generally consistent with previously published work. However, N₂O emissions from the two Scottish soils were substantially more sensitive to inorganic N fertiliser application at 80% water holding capacity than the other soils, with the N₂O emissions being up to 107 times higher than the other studied soils.     

Immunostimulatory effects of dietary poly‐β‐hydroxybutyrate in European sea bass postlarvae

The stable production of high‐quality fry in marine aquaculture is still hampered by unpredictable mortality caused by infectious diseases during larval rearing. Consequently, the development of new biocontrol agents is crucial for a viable aquaculture industry. The bacterial energy storage compound poly‐β‐hydroxybutyrate (PHB) has been shown to exhibit beneficial properties on aquatic organisms such as enhanced survival, growth, disease resistance and a controlling effect on the gastrointestinal microbiota. However, the effect of PHB on the developing immune system of fish larvae has so far not been investigated. In this study, the effect of feeding PHB‐enriched Artemia nauplii on survival, growth and immune response in European sea bass (Dicentrarchus labrax) postlarvae was examined. Amorphous PHB was administered to 28‐day‐old sea bass postlarvae over a period of 10 days. The survival and growth performance were monitored, and the expression of 29 genes involved in immunity, growth, metabolism and stress‐response was measured. While the expression of the insulin‐like growth factor 1 (igf1), an indicator of relative growth, was upregulated in response to feeding PHB, the larval survival and growth performance remained unaffected. After 10 days of PHB treatment, the expression of the antimicrobial peptides dicentracin (dic) and hepcidin (hep) as well as mhc class IIa and mhc class IIb was elevated in the PHB fed postlarvae. This indicates that PHB is capable of stimulating the immune system of fish early life stages, which may be the cause of the increased resistance to diseases and robustness observed in previous studies.     

Comparative Study of Predominant Phytochemical Compounds and Proapoptotic Potential of Broccoli Sprouts and Florets

The study compares lyophilized broccoli sprouts and florets in terms of their chemical composition, cytotoxic and proapoptotic potential against hepatocellular carcinoma HepG2, colorectal cancer SW480, and skin fibroblast BJ cells. Sinapic and isochlorogenic acids were predominant phenolics in the sprouts and florets, respectively. The amount of sulforaphane in the sprouts was significantly higher vs. florets. Oleic and linoleic acids dominated in the sprouts, while caproic, stearic and oleic acids in the florets. Broccoli sprouts were selectively cytotoxic on HepG2 and SW480 cells, with proapoptotic effect for the latter, while the florets were less selective, but more active, with profound proapoptotic effect for HepG2 cells (77.4%). Thus, lyophilized broccoli sprouts may be effectively used in dietary chemoprevention.