Silencing the gip gene of Phytophthora cinnamomi by iRNA and studying the subcellular localization of GIP and NPP1 proteins

Ink Disease is considered one of the most important causes of the decline of chestnut orchards. The break in yield of Castanea sativa Mill is caused by two species: Phytophthora cinnamomi and Phytophthora cambivora, being the first one the foremost pathogen of ink disease in Portugal. P. cinnamomi is one of the most aggressive and widespread plant pathogen with nearly 1,000 host species. This oomycete causes enormous economic losses and it is responsible for the decline of many plant species in Europe and worldwide. Up to now no efficient treatments are available to fight these pathogens. Because of the importance of chestnut at economical and ecological levels, especially in Portugal, it becomes essential to explore the molecular mechanisms that determine the interaction between Phytophthora species and host plants through the study of proteins GIP (glucanase inhibitor protein) and NPP1 (necrosis-inducing Phytophthora protein 1) produced by P. cinnamomi during the infection. The technique of RNA interference was used to knockdown the gip gene of P. cinnamomi. Transformants obtained with the silenced gene have been used to infect C. sativa, in order to determine the effect of gene silencing on the plant phenotype. To know more about the function of GIP and NPP1 involved in the mechanism of infection, the ORF’s of gip and npp1 genes have been cloned to the pTOR-eGFP vector for a future observation of P. cinnamomi transformants with fluorescent microscopy and determination of the subcellular localization. Moreover the prediction by bioinformatics tools indicates that both GIP and NPP1 proteins are secreted. The results allow to predict the secretory destination of both GIP and NPP1 proteins and confirm RNAi as a potential alternative biological tool in the control and management of P. cinnamomi. Keywords:

Alzheimer’s electroencephalogram event scalp and source localization

Alzheimer’s disease is the most common cause of dementia which causes a progressive and irreversible impairment of several cognitive functions. The aging population has been increasing significantly in recent decades and this disease affects mainly the elderly. Its diagnostic accuracy is relatively low and there is not a biomarker able to detect AD without invasive tests. Despite the progress in better understanding the disease there remains no prospect of cure at least in the near future. The electroencephalogram (EEG) test is a widely available technology in clinical settings. It may help diagnosis of brain disorders, once it can be used in patients who have cognitive impairment involving a general decrease in overall brain function or in patients with a located deficit. This study is a new approach to improve the scalp localization and the detection of brain anomalies (EEG temporal events) sources associated with AD by using the EEG.

ViDRILO: The Visual and Depth Robot Indoor Localization with Objects information dataset

In this article we describe a semantic localization dataset for indoor environments named ViDRILO. The dataset provides five sequences of frames acquired with a mobile robot in two similar office buildings under different lighting conditions. Each frame consists of a point cloud representation of the scene and a perspective image. The frames in the dataset are annotated with the semantic category of the scene, but also with the presence or absence of a list of predefined objects appearing in the scene. In addition to the frames and annotations, the dataset is distributed with a set of tools for its use in both place classification and object recognition tasks. The large number of labeled frames in conjunction with the annotation scheme make this dataset different from existing ones. The ViDRILO dataset is released for use as a benchmark for different problems such as multimodal place classification and object recognition, 3D reconstruction or point cloud data compression.

Glycogen Synthase Kinase 3 Influences Cell Motility and Chemotaxis by Regulating Phosphatidylinositol 3 Kinase Localization in Dictyostelium discoideum

Glycogen Synthase Kinase 3 (GSK3), a serine/threonine kinase initially characterized in the context of glycogen metabolism, has been repeatedly realized as a multitasking protein that can regulate numerous cellular events in both metazoa and protozoa. I recently found GSK3 plays a role in regulating chemotaxis, a guided cell movement in response to an external chemical gradient, in one of the best studied model systems for chemotaxis - Dictyostelium discoideum. It was initially found that comparing to wild type cells, gsk3- cells showed aberrant chemotaxis with a significant decrease in both speed and chemotactic indices. In Dictyostelium, phosphatidylinositol 3,4,5-triphosphate (PIP3) signaling is one of the best characterized pathways that regulate chemotaxis. Molecular analysis uncovered that gsk3- cells suffer from high basal level of PIP3, the product of PI3K. Upon chemoattractant cAMP stimulation, wild type cells displayed a transient increase in the level of PIP3. In contrast, gsk3- cells exhibited neither significant increase nor adaptation. On the other hand, no aberrant dynamic of phosphatase and tensin homolog (PTEN), which antagonizes PI3K function, was observed. Upon membrane localization of PI3K, PI3K become activated by Ras, which will in turn further facilitate membrane localization of PI3K in an F-Actin dependent manner. The gsk3- cells treated with F-Actin inhibitor Latrunculin-A showed no significant difference in the PIP3 level. I also showed GSK3 affected the phosphorylation level of the localization domain of PI3K1 (PI3K1-LD). PI3K1-LD proteins from gsk3- cells displayed less phosphorylation on serine residues compared to that from wild type cells. When the potential GSK3 phosphorylation sites of PI3K1-LD were substituted with aspartic acids (Phosphomimetic substitution), its membrane localization was suppressed in gsk3- cells. When these serine residues of PI3K1-LD were substituted with alanine, aberrantly high level of membrane localization of the PI3K1-LD was monitored in wild type cells. Wild type, phosphomimetic, and alanine substitution of PI3K1-LD fused with GFP proteins also displayed identical localization behavior as suggested by the cell fraction studies. Lastly, I identified that all three potential GSK3 phosphorylation sites on PI3K1-LD could be phosphorylated in vitro by GSK3.

Localization and edafo-climate characteristics of Sete Lagoas and Janaúba (MG) specific sites-Embrapa Maize and Sorghum.

2008

Construction defect reporting using mobile and digital workbench technologies

Sending data between the construction site and an off-site design office is one of the more problematic areas in information technology for construction automation, particularly for construction defect management. The aim of this research is to investigate how mobile computing and new forms of human-computer interaction can be brought to bear on specific problems in construction management. The construction defect reporting system is one such application. Combining mobile and wireless computing technologies with a digital workbench, we have developed a system to facilitate remote telecollaboration between a construction site and an off-site engineering office. The application reported in this paper demonstrates how construction defect reporting can be streamlined by field collection of construction defect information using a mobile device and visualising the defect in a CAD model on a digital workbench in an engineering office. This paper reports on the design of the system and our tests of sending images from the construction site to the engineer’s office and positional accuracy of GPS for localization of the defect.

Stem cell regulatory gene expression in human adult dental pulp and periodontal ligament cells undergoing odontogenic/osteogenic differentiation

Introduction During development and regeneration, odontogenesis and osteogenesis are initiated by a cascade of signals driven by several master regulatory genes. Methods In this study, we investigated the differential expression of 84 stem cell–related genes in dental pulp cells (DPCs) and periodontal ligament cells (PDLCs) undergoing odontogenic/osteogenic differentiation. Results Our results showed that, although there was considerable overlap, certain genes had more differential expression in PDLCs than in DPCs. CCND2, DLL1, and MME were the major upregulated genes in both PDLCs and DPCs, whereas KRT15 was the only gene significantly downregulated in PDLCs and DPCs in both odontogenic and osteogenic differentiation. Interestingly, a large number of regulatory genes in odontogenic and osteogenic differentiation interact or crosstalk via Notch, Wnt, transforming growth factor β (TGF-β)/bone morphogenic protein (BMP), and cadherin signaling pathways, such as the regulation of APC, DLL1, CCND2, BMP2, and CDH1. Using a rat dental pulp and periodontal defect model, the expression and distribution of both BMP2 and CDH1 have been verified for their spatial localization in dental pulp and periodontal tissue regeneration. Conclusions This study has generated an overview of stem cell–related gene expression in DPCs and PDLCs during odontogenic/osteogenic differentiation and revealed that these genes may interact through the Notch, Wnt, TGF-β/BMP, and cadherin signalling pathways to play a crucial role in determining the fate of dental derived cell and dental tissue regeneration. These findings provided a new insight into the molecular mechanisms of the dental tissue mineralization and regeneration

Computational approach to localization using global energy minimization

Damage localization induced by strain softening can be predicted by the direct minimization of a global energy function. This article concerns the computational strategy for implementing this principle for softening materials such as concrete. Instead of using heuristic global optimization techniques, our strategies are a hybrid of local optimization methods with a path-finding approach to ensure a global optimum. With admissible nodal displacements being independent variables, it is easy to deal with the geometric (mesh) constraint conditions. The direct search optimization methods recover the localized solutions for a range of softening lattice models which are representative of quasi-brittle structures