Early development of Moniliophthora perniciosa basidiomata and developmentally regulated genes

<p>Abstract</p> <p>Background</p> <p>The hemibiotrophic fungus <it>Moniliophthora perniciosa </it>is the causal agent of Witches' broom, a disease of <it>Theobroma cacao</it>. The pathogen life cycle ends with the production of basidiocarps in dead tissues of the infected host. This structure generates millions of basidiospores that reinfect young tissues of the same or other plants. A deeper understanding of the mechanisms underlying the sexual phase of this fungus may help develop chemical, biological or genetic strategies to control the disease.</p> <p>Results</p> <p>Mycelium was morphologically analyzed prior to emergence of basidiomata by stereomicroscopy, light microscopy and scanning electron microscopy. The morphological changes in the mycelium before fructification show a pattern similar to other members of the order <it>Agaricales</it>. Changes and appearance of hyphae forming a surface layer by fusion were correlated with primordia emergence. The stages of hyphal nodules, aggregation, initial primordium and differentiated primordium were detected. The morphological analysis also allowed conclusions on morphogenetic aspects. To analyze the genes involved in basidiomata development, the expression of some selected EST genes from a non-normalized cDNA library, representative of the fruiting stage <it>of M. perniciosa</it>, was evaluated. A macroarray analysis was performed with 192 selected clones and hybridized with two distinct RNA pools extracted from mycelium in different phases of basidiomata formation. This analysis showed two groups of up and down-regulated genes in primordial phases of mycelia. Hydrophobin coding, glucose transporter, Rho-GEF, Rheb, extensin precursor and cytochrome p450 monooxygenase genes were grouped among the up-regulated. In the down-regulated group relevant genes clustered coding calmodulin, lanosterol 14 alpha demethylase and PIM1. In addition, 12 genes with more detailed expression profiles were analyzed by RT-qPCR. One aegerolysin gene had a peak of expression in mycelium with primordia and a second in basidiomata, confirming their distinctiveness. The number of transcripts of the gene for plerototolysin B increased in reddish-pink mycelium and indicated an activation of the initial basidiomata production even at this culturing stage. Expression of the glucose transporter gene increased in mycelium after the stress, coinciding with a decrease of adenylate cyclase gene transcription. This indicated that nutrient uptake can be an important signal to trigger fruiting in this fungus.</p> <p>Conclusion</p> <p>The identification of genes with increased expression in this phase of the life cycle of <it>M. perniciosa </it>opens up new possibilities of controlling fungus spread as well as of genetic studies of biological processes that lead to basidiomycete fruiting. This is the first comparative morphologic study of the early development both <it>in vivo </it>and <it>in vitro </it>of <it>M. perniciosa </it>basidiomata and the first description of genes expressed at this stage of the fungal life cycle.</p

Impact of Alignment Edits on the Quality of Experience of 360&#x00B0; Videos

Optimizing user quality of experience (QoE) for 360&#x00B0; videos faces two major roadblocks: inaccurate viewport prediction and viewers missing the plot of a story. To tackle these issues simultaneously, alignment edits have emerged as a promising solution. These edits, also known as &#x201C;re-targeting edits,&#x201D; work by aligning the user&#x2019;s field of view with a specific region of interest in the video content. Despite their potential benefits, there is limited knowledge about the actual impacts of alignment edits on user experience (UX). Therefore, we conducted subjective experiments following ITU-T P.919 methodology to explore their effects on QoE. We proposed an alignment edit based on gradual rotation of the 360&#x00B0; frame, aiming to replicate natural viewing behavior. We tested this approach under various conditions and thoroughly analyzed its impacts using both head motion data and feedback from observers, focusing on their sense of presence, comfort, and perceived experience. The results of our experiments are encouraging. Our proposed gradual alignment technique achieved a level of comfort and presence comparable to that of instant edits. Furthermore, all alignment edits tested led to a noticeable reduction in head movement speed after the edit, affirming their potential utility for on-demand video streaming. Notably, the gradual edits, in particular, induced a significant reduction of approximately 8&#x0025; in head movement speeds when compared to the instant alignment technique. These findings shed light on the positive effects of alignment edits on user experience and firmly establish the viability of the proposed gradual alignment technique to enhance QoE during video consumption

3D in vitro models for skin and oral mucosa repair: a methodological study and bibliometric analysis

In vitro studies are widely used in pharmacological and biological fields as the first step when devel-oping new approaches and technologies. The main goal is to be a precursor of clinical research, which makes the ability to get close to the clinical context extremely important. For skin and oral mucosa, 2D cell culture techniques are the reference standard, being specially used for morphological assays, mo-lecular interactions, drug screening, and wound healing. Nevertheless, the skin consists of multiple layers made up of different types of cells. When it comes to skin functions, tissues and cells interac-tions are essential, challenging the current monolayer strategies besides punctuating several limitations. In this view, complex 3D models, such as culture systems, bioprints, and organoids, have been sug-gested to improve the quality and reliability of in vitro analysis. Reliable three-dimensional skin equiv-alents would enable access to unique properties of skin, such as wound healing, epithelialization, for-mation of hair follicles and appendages, barrier function and senescence, as well as complex skin dis-eases including psoriasis, dermatitis, infections, and cancer. Also, it would contribute to transplanta-tion therapy in cases of serious injuries such as burn wounds and chronic diabetic. Thus, the estab-lishment of skin equivalents might represent a huge advancement in tissue engineering, providing re-search models and skin substitutes to be applied for industrial, pre-clinical, and clinical purposes. Alt-hough significant progress has been made in this field, a standard and reproducible protocol for artifi-cial skin substitutes is still a challenge. Different in development protocols are available but most of them are not fully standardized and adequate. This methodological systematic review aims to assess all the studies using 3D models for skin/oral mucosa and summarize the available approaches to suggest the best standard and reliable options until this moment, in addition to highlight current challenges and future perspectives

Sialyl Tn-expressing bladder cancer cells induce a tolerogenic phenotype in innate and adaptive immune cells

This work was supported by the Portuguese Foundation for Science and Technology (FCT) - PTDC/SAU-MII/67561/2006 and Premio Santander Totta - UNL (Paula A. Videira), LPCC/Pfizer2011 (Mylene A. Carrascal), SFRH/BPD/21619/2005 (M. Guadalupe Cabral), SFRH/BD/81860/2011 (Mariana Silva), SFRH/BD/45120/2008 (Paulo F. Severino) and SFRH/BPD/ 66288/2009 (Jose Alexandre Ferreira). FCT is co-financed by European Social Fund (ESF) under Human Potential Operation Programme (POPH) from National Strategic Reference Framework (NSRF).) and European Union, QREN, FEDER, COMPETE, for funding the Organic Chemistry Research Unit (QOPNA) (project PEst-C/QUI/UI0062/2013; FCOMP-01-0124-FEDER-037296).Despite the wide acceptance that glycans are centrally implicated in immunity, exactly how they contribute to the tilt immune response remains poorly defined. In this study, we sought to evaluate the impact of the malignant phenotype-associated glycan, sialyl-Tn (STn) in the function of the key orchestrators of the immune response, the dendritic cells (DCs). In high grade bladder cancer tissue, the STn antigen is significantly overexpressed and correlated with the increased expression of ST6GALNAC1 sialyltransferase. Bladder cancer tissue presenting elevated expression of ST6GALNAC1 showed a correlation with increased expression of CD1a, a marker for bladder immature DCs and showed concomitant low levels of Th1-inducing cytokines IL-12 and TNF-α. Invitro, human DCs co-incubated with STn+ bladder cancer cells, had an immature phenotype (MHC-IIlow, CD80low and CD86low) and were unresponsive to further maturation stimuli. When contacting with STn+ cancer cells, DCs expressed significantly less IL-12 and TNF-α. Consistent with a tolerogenic DC profile, T cells that were primed by DCs pulsed with antigens derived from STn+ cancer cells were not activated and showed a FoxP3high IFN-γlow phenotype. Blockade of STn antigens and of STn+ glycoprotein, CD44 and MUC1, in STn+ cancer cells was able to lower the induction of tolerance and DCs become more mature.Overall, our data suggest that STn-expressing cancer cells impair DC maturation and endow DCs with a tolerogenic function, limiting their capacity to trigger protective anti-tumour T cell responses. STn antigens and, in particular, STn+ glycoproteins are potential targets for circumventing tumour-induced tolerogenic mechanisms.publishersversionpublishe

Botanical secondary metabolites as alternatives for pest management in Cuba

Plants have historically been used in agricultural practices in Cuba; nevertheless, the Cuban flora has not yet been fully studied as a potential source of pesticides, partly due to its great diversity. At the National Centre for Animal and Plant Health (CENSA), systematic research on more than 100 plant species belonging to several families has been conducted in recent years. The protocol involves the establishment of bioassay conditions, the isolation and characterization of bioactive compounds, assessment of compatibility with other biological control agents, mode of action studies and the semi-synthesis of analogues using biotechnological techniques. Among the secondary metabolites studied, essential oils obtained from plants belonging to the families Piperaceae, Lamiaceae, Apiaceae, Rutaceae, and Myrtaceae stood out as a promising group due to their efficacy and spectrum of action under laboratory and semi-controlled conditions. As botanical pesticides, the main areas of use may include protected crops, nurseries, seed treatments in protected and field-grown crops, and in stored product pest management. The use of known botanicals and the identification of local candidates for developing phytosanitary products offer alternatives that may combine efficiency and safety for pest management in Cuban agriculture.Plants have historically been used in agricultural practices in Cuba; nevertheless, the Cuban flora has not yet been fully studied as a potential source of pesticides, partly due to its great diversity. At the National Centre for Animal and Plant Health (CENSA), systematic research on more than 100 plant species belonging to several families has been conducted in recent years. The protocol involves the establishment of bioassay conditions, the isolation and characterization of bioactive compounds, assessment of compatibility with other biological control agents, mode of action studies and the semi-synthesis of analogues using biotechnological techniques. Among the secondary metabolites studied, essential oils obtained from plants belonging to the families Piperaceae, Lamiaceae, Apiaceae, Rutaceae, and Myrtaceae stood out as a promising group due to their efficacy and spectrum of action under laboratory and semi-controlled conditions. As botanical pesticides, the main areas of use may include protected crops, nurseries, seed treatments in protected and field-grown crops, and in stored product pest management. The use of known botanicals and the identification of local candidates for developing phytosanitary products offer alternatives that may combine efficiency and safety for pest management in Cuban agricultur