Summary In this study, we compare the histological localisation of phenols and β‐glucan in six cereal cross sections. The concentrations of β‐glucan and total phenols in whole grain flour (WGF) were also investigated to provide their nutrient content in grams. Cross sections of durum, soft, einkorn and emmer wheat, oats and barley were stained with Azan Trichrome (ATH) and periodic acid–Schiff (PAS) to characterise the morphology and cytohistology of the living cells in the aleurone, subaleurone and germ, as well as the a‐nucleated cells in the starchy endosperm. Phenol localisation in the outer layers was performed by fluorescence microscopy due to the presence of ferulic acid. β‐Glucan, examined with Calcofluor White staining, was found to be located primarily in the subaleurone layer and starchy endosperm of oats and barley. Our findings shed light on why WGF nutritional value is markedly reduced when the bran is removed through milling.
Psoriasis is a chronic skin disease associated with deregulated activation of immune cells and keratinocytes. In this study, we used the imiquimod (IMQ)-induced mouse model of psoriasis to dissect better the contribution of hematopoietic and skin-resident stromal cells to psoriasis development. The comparison of disease development in mice carrying the hematopoietic cell-specific deletion of MyD88 ( mice) with mice carrying the total MyD88 deficiency ( mice), we show that the progression of skin and systemic inflammation, as well as of epidermal thickening, was completely dependent on MyD88 expression in hematopoietic cells. However, both mouse strains developed some degree of epidermal thickening during the initial stages of IMQ-induced psoriasis, even in the absence of hematopoietic cell activation and infiltration into the skin, suggesting a contribution of MyD88-independent mechanisms in skin-resident stromal cells. With the use of conditional knockout mouse strains lacking MyD88 in distinct lineages of myeloid cells ( and mice), we report that MyD88 signaling in monocytes and Mϕ, but not in neutrophils, plays an important role in disease propagation and exacerbation by modulating their ability to sustain γδ T cell effector functions via IL-1β and IL-23 production. Overall, these findings add new insights into the specific contribution of skin-resident stromal vs. hematopoietic cells to disease initiation and progression in the IMQ-induced mouse model of psoriasis and uncover a potential novel pathogenic role for monocytes/Mϕ to psoriasis development.
The seed morphology of three Pseudocereal Grains (PSCg), i.e. quinoa (Chenopodium quinoa Willd, Chenopodiaceae), buckwheat (Fagopyrum esculentum Moench, Polygonaceae) and amaranth (Amaranthus caudatus L., Amaranthaceae) was studied by light microscopy (LM) and Environmental Scanning Electron Microscopy coupled with Energy Dispersive Spectroscopy (ESEM-EDS). LM was used with visible light to evaluate either unstained sections or sections stained with Azan mixture and with fluorescent light. The aim of the study was to compare the architecture of the three seeds in order to connect their morphology with nutrient localization. The Azan staining allowed for the visualization of the seed coat, the embryo - with its shoot apical meristem - and the radicle cell layers, whereas the use of fluorescent microscopy identified the cells rich in phenolic compounds. Finally, the ESEM-EDS analysis revealed that the seed coat of the quinoa was thinner than that of amaranth or buckwheat. In all PSCg, starch granules appeared to be located in large polygonal cells, surrounded by a thin cell wall. Several globoids of proteins were observed in the embryo cells. In the radicle section, the vascular bundles of the procambium were evident, while Amaranth only showed a consistent layer of calcium crystals, located between the embryo and the perysperm. The morphological differences of the three PSCg were discussed in the context of their structural resistance to processing technologies which impact on nutritional value of derived foods.
In an effort to find alternatives to study in vivo the so-called New Psychoactive Substances (NPS), the present work was undertaken to investigate the use of zebrafish larvae as animal model in pharmaco-toxicology, providing behavioural and metabolism information. For this purpose, fentanyl, the progenitor of an extremely dangerous group of NPS, was administered at different doses to zebrafish larvae (1, 10, 50, 100 µM) in comparison to mice (0.1, 1, 6, 15 mg/kg), as a well-established animal model. A behavioural assay was performed at the time of the peak effect of fentanyl, showing that the results in larvae are consistent with those observed in mice. On the other hand, several morphological abnormalities (namely yolk sac edema, abnormal pericardial edema, jaw defect and spinal curvature) were found in larvae mostly at high fentanyl doses (50, 100 µM). Larva extract and mice urine were analyzed by using liquid chromatography coupled to high resolution mass spectrometry to identify the metabolic pathways of fentanyl. The main metabolites detected were norfentanyl and hydroxyfentanyl in both the tested models. In conclusion, the present study provides evidence that fentanyl effects on zebrafish larvae and metabolism are similar to rodents and consequently support the hypothesis of using zebrafish larvae as a suitable rapid screening tool to investigate new drugs, and particularly NPS.
The present study investigated the morphology of fresh and brine-cured table olives (TOs) as well as the changes that occur when drupes are attacked by the fruit fly Bactrocera oleae. Morphological analyses were performed using light microscopy (LM) and environmental scanning electron microscopy coupled with energy dispersive spectroscopy (ESEM-EDS). The LM analysis was carried out with visible light to evaluate sections stained with either PAS or Azan mixtures as well as unstained sections observed at fluorescence microscopy. The results of the analyses showed that: i) Azan and PAS staining played a useful complementary role, increasing the information provided by the histological analysis. Indeed, in both fresh and brine-cured TOs, epidermal layers and mesocarpal cells were clearly revealed, including sclereid cells. The histological analysis allowed also to identifying the presence of secoiridoid-biophenols (seco-BPs) in both cell walls and vacuoles, as well as in the drupe regions that had been attacked by fruit flies, where they were found at higher concentrations; ii) in fresh and brine-cured olives, the excitation at 480 nm revealed the distribution of the fluorophores, among which the seco-BP are enclosed; iii) the ESEM-EDS analysis revealed the natural morphology of fresh olives, including the dimensions of their cell layers and the size and depth of the mechanical barriers of suberized or necrotic cells around the larva holes. In addition, the elemental composition of regions of interest of the drupe was determined in fresh and brine-cured TOs. The results highlighted the effectiveness of combined use of LM and ESEM-EDS in order to obtain a picture, as complete as possible, of the structural morphology of TOs. Such analytical combined approach can be used to support multidisciplinary studies aimed at the selection of new cultivars more resistant to fly attack.
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