Identification of novel regulators of COP1-controlled morphogenesis in Arabidopsis thaliana

In Arabidopsis thaliana, COP1 is an essential element of light signal transduction acting downstream of photoreceptors and upstream of light-regulated gene expression. The COP1 protein acts as part of an E3 ligase complex to suppress photomorphogenic gene expression by ubiquitin-dependent degradation of light-regulated transcription factors. In dark-grown seedlings, the repression of photomorphogenesis involves the inhibition of hypocotyl growth, anthocyanin accumulation, expression of light-responsive genes, differentiation of etioplasts and prevention of apical hook formation. Loss of COP1 function leads to a pleiotropic phenotype comprising of constitutive photomorphogenesis in the dark and resulting in a post-germination growth arrest. The vegetative growth arrest of cop1 mutants, a possible role of COP1 concerning the cell cycle and the molecular factors regulating the nucleocytoplasmic partitioning of COP1 exemplify aspects of COP1 function and regulation that are poorly understood until now. This work aimed at the identification of regulators of COP1-controlled morphogenesis to contribute to a better dissection of the latter. In yeast two hybrid screenings (YTH) 32 new interaction candidates for COP1 were identified and a purpose oriented selection was performed. In order to select a putative regulator of COP1, all COP1 and additional DET1 interaction candidates were integrated in a network of published interactors. Out of the network and the screening results PAP2 (PRODUCTION OF ANTHOCYAN PIGMENT) was identified and selected as a putative new target. MID (= MIDGET) was selected as a putative new regulator of COP1, respectively. MID is a part of the topoisomerasis VI (TOPOVI) complex that is needed to complete more than two endocycles in plant cells. This work provides evidence for a physical interaction of MID and PAP2 with COP1. In addition, a new YTH-based domain mapping method was developed and used to identify so far unknown domains of PAP2 for the interaction with COP1 and for COP1 for the interaction with MID and TOPOVI components. Similar to cop1, mid and topoisomerase VI mutants exhibited all aspects of constitutive pohotomorphogenesis in the dark. Double mutant analysis indicated that MID is not a target of COP1. In infiltrated leaves of Nicotiana benthamina, the presence of MID is needed for COP1 to form a high number of subnuclear foci. MID and the TOPOVI were shown to be essential regulators of COP1 function probably by stabilising COP1 and thereby adding a new cell-cycle related factor to the regulation of COP1 activity. The functional relevance of the MID-COP1 interaction was proven by analysing phenotypes of the single mutants and genetic interaction. First evidence positioning MID in a SPA1 and phyA-dependent complex or pathway were obtained by the verification of the SPA1-MID interaction via BiFC, co-purification of MID with phyA and analysis of the protein stability of MID depending on light quality. Finally it was found that mid and topoVI mutants phenocopy det1-1 mutants and overexpressor lines of the C-termini of CRY1 and CRY2, possibly providing a new link to crosstalk between red and blue light mediated signaling

Development of multifunction ladder

The objective of this thesis is to develop and fabricate the multifunction ladder and to minimize the manufacturing cost. Ladder is equipment that can easier consumer’s daily activities at higher place. It is stable and safe to use. The concept of this project is based on student’s creativity. The characteristics of material to fabricate this ladder are lightweight, longer life span, corrosion resistance and can take maximum load. Thus, the suitable material that accomplished those characteristics is aluminium alloy. In this thesis, we’ll also be having more to the fabrication of this ladder

Do nursing and pharmacy students practice what they preach on safe drug storage and disposal? A cross-sectional study

Background: Research has confirmed a lack of knowledge regarding the risks of unused medications including diversion, misuse, or accidental overdose among health care professionals (Abdulmajeed, 2020). Nurses and pharmacists are often who patients interact with the most regarding medications; therefore, early education on proper storage and disposal is vital (Bowen, Rotz, Patterson, & Sen, 2017; Celio, Ninane, Bugnon, & Schneider, 2018). Objectives: The study\u27s objective is to explore safe drug storage and disposal knowledge, attitudes, and practices of professional pharmacy (Pharm.D.) and nursing students. Design: This research is an exploratory cross-sectional study from May to September 2019. Methods: An anonymous online survey was administered to a purposive sample of Pharm.D. and nursing students who were 18 years and older and enrolled in the site\u27s accredited Pharm.D. and nursing programs (N = 210). Responses were analyzed using descriptive statistics. Results: Common disposal methods reported by students of their personal medications such as pills and liquids included discarding medications with the household trash (range 30% to 55%) and medication disposal products/bag (range 19% to 28%). More than half of the participants (50.4%) had unused prescription medication at home, 35% kept the medication for later use, and almost 20% of the participants reported sharing personal medications with others. Conclusion: Although the majority of student participants had adequate knowledge of the appropriate methods for safe drug disposal, few reported using them for their own personal medications. The findings suggest there is a disconnect between the participants\u27 knowledge of the appropriate methods of safe drug storage and disposal in a professional setting and their own practices. Further research is needed to explore and address the reasons for this disconnect. Additionally, findings from this research will assist in the development of and/or the improvement of interdisciplinary educational materials among pharmacy and nursing students

Ontologies for increasing the FAIRness of plant research data

The importance of improving the FAIRness (findability, accessibility, interoperability, reusability) of research data is undeniable, especially in the face of large, complex datasets currently being produced by omics technologies. Facilitating the integration of a dataset with other types of data increases the likelihood of reuse, and the potential of answering novel research questions. Ontologies are a useful tool for semantically tagging datasets as adding relevant metadata increases the understanding of how data was produced and increases its interoperability. Ontologies provide concepts for a particular domain as well as the relationships between concepts. By tagging data with ontology terms, data becomes both human and machine interpretable, allowing for increased reuse and interoperability. However, the task of identifying ontologies relevant to a particular research domain or technology is challenging, especially within the diverse realm of fundamental plant research. In this review, we outline the ontologies most relevant to the fundamental plant sciences and how they can be used to annotate data related to plant-specific experiments within metadata frameworks, such as Investigation-Study-Assay (ISA). We also outline repositories and platforms most useful for identifying applicable ontologies or finding ontology terms.Comment: 34 pages, 4 figures, 1 table, 1 supplementary tabl

Metabolic shift toward oxidative phosphorylation in docetaxel resistant prostate cancer cells

Drug resistance of cancer cells is recognized as the primary cause of failure of chemotherapeutic treatment in most human cancers. Growing evidences support the idea that deregulated cellular metabolism is linked to such resistance. Indeed, both components of the glycolytic and mitochondrial pathways are involved in altered metabolism linked to chemoresistance of several cancers. Here we investigated the drug-induced metabolic adaptations able to confer advantages to docetaxel resistant prostate cancer (PCa) cells. We found that docetaxel-resistant PC3 cells (PC3-DR) acquire a pro-invasive behavior undergoing epithelial-to-mesenchymal-transition (EMT) and a decrease of both intracellular ROS and cell growth. Metabolic analyses revealed that PC3-DR cells have a more efficient respiratory phenotype than sensitive cells, involving utilization of glucose, glutamine and lactate by the mitochondrial oxidative phosphorylation (OXPHOS). Consequently, targeting mitochondrial complex I by metformin administration, impairs proliferation and invasiveness of PC3-DR cells without effects on parental cells. Furthermore, stromal fibroblasts, which cause a "reverse Warburg" phenotype in PCa cells, reduce docetaxel toxicity in both sensitive and resistant PCa cells. However, re-expression of miR-205, a microRNA strongly down-regulated in EMT and associated to docetaxel resistance, is able to shift OXPHOS to a Warburg metabolism, thereby resulting in an elevated docetaxel toxicity in PCa cells. Taken together, these findings suggest that resistance to docetaxel induces a shift from Warburg to OXPHOS, mandatory for conferring a survival advantage to resistant cells, suggesting that impairing such metabolic reprogramming could be a successful therapeutic approach.Associazione Italiana Ricerca sul Cancro (AIRC), Istituto Toscano Tumori and Regione Toscan

Two structurally distinct domains of the nucleoporin Nup170 cooperate to tether a subset of nucleoporins to nuclear pores

How individual nucleoporins (Nups) perform their role in nuclear pore structure and function is largely unknown. In this study, we examined the structure of purified Nup170 to obtain clues about its function. We show that Nup170 adopts a crescent moon shape with two structurally distinct and separable domains, a β-propeller N terminus and an α-solenoid C terminus. To address the individual roles of each domain, we expressed these domains separately in yeast. Notably, overexpression of the Nup170 C domain was toxic in nup170Δ cells and caused accumulation of several Nups in cytoplasmic foci. Further experiments indicated that the C-terminal domain anchors Nup170 to nuclear pores, whereas the N-terminal domain functions to recruit or retain a subset of Nups, including Nup159, Nup188, and Pom34, at nuclear pores. We conclude that Nup170 performs its role as a structural adapter between cytoplasmically oriented Nups and the nuclear pore membrane

CCR7 as a novel therapeutic target in t-cell PROLYMPHOCYTIC leukemia

T-cell prolymphocytic leukemia (T-PLL) is a poor prognostic disease with very limited options of efficient therapies. Most patients are refractory to chemotherapies and despite high response rates after alemtuzumab, virtually all patients relapse. Therefore, there is an unmet medical need for novel therapies in T-PLL. As the chemokine receptor CCR7 is a molecule expressed in a wide range of malignancies and relevant in many tumor processes, the present study addressed the biologic role of this receptor in T-PLL. Furthermore, we elucidated the mechanisms of action mediated by an anti-CCR7 monoclonal antibody (mAb) and evaluated whether its anti-tumor activity would warrant development towards clinical applications in T-PLL. Our results demonstrate that CCR7 is a prognostic biomarker for overall survival in T-PLL patients and a functional receptor involved in the migration, invasion, and survival of leukemic cells. Targeting CCR7 with a mAb inhibited ligand-mediated signaling pathways and induced tumor cell killing in primary samples. In addition, directing antibodies against CCR7 was highly effective in T-cell leukemia xenograft models. Together, these findings make CCR7 an attractive molecule for novel mAb-based therapeutic applications in T-PLL, a disease where recent drug screen efforts and studies addressing new compounds have focused on chemotherapy or small molecules.Peer reviewe

Cadmium interferes with auxin physiology and lignification in poplar

Cadmium (Cd) is a phytotoxic heavy metal that causes rapid growth reduction. To investigate if Cd interferes with the metabolism of auxin, a major growth hormone in plants, poplars (Populus×canescens) expressing a heterologous GH3::GUS reporter gene were exposed to 50 μM Cd in hydroponic solutions. Growth, photosynthetic performance, lignification, peroxidase activity, auxin concentration, and GUS staining were determined in order to record the activities of GH3 enzymes in the stem apex, the elongation zone, wood in the zone of radial growth, and in roots. Cd-induced growth reductions were tissue-specific decreasing in the order: roots>wood>shoot elongation and leaf initiation, whereas Cd concentrations increased in the order: leaves<wood<roots. Cd almost abolished the GH3 signal in the stem apex but caused strong increases in the vascular system of roots as well as in parenchymatic cells in the xylem. These changes were accompanied by increases in lignin and peroxidase activities and decreases in auxin concentrations. Since GH3 enzymes remove auxin from the active pool by conjugation and act as mediators between growth and defence, our data suggest that Cd stress triggered increases in GH3 activities which, in turn, depleted auxin in wood and thereby shunted the metabolism to enhanced formation of lignin