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1.
J Vis Exp ; (191)2023 01 20.
Article in English | MEDLINE | ID: mdl-36744777

ABSTRACT

Stomata are small pores on the surface of land plants that are involved in gas exchange and water vapor release, and their function is critical for plant productivity and survival. As such, understanding the mechanisms by which stomata develop and pattern has tremendous agronomic value. This paper describes two phenotypic methods using Arabidopsis cotyledons that can be used to characterize the genes controlling stomatal development and patterning. Presented first are procedures for analyzing the stomatal phenotypes using toluidine blue O-stained cotyledons. This method is fast and reliable and does not require the use of epidermal peels, which are widely used for phenotypic analyses but require specialized training. Due to the presence of multiple cysteine residues, the identification and generation of bioactive EPF peptides that have a role in stomatal development have been challenging. Thus, presented second is a procedure used to identify stomatal ligands and monitor their biological activity by bioassays. The main advantage of this method is that it produces reproducible data relatively easily while reducing the amount of peptide solution and the time required to characterize the role of the peptides in controlling stomatal patterning and development. Overall, these well-designed protocols enhance the efficiency of studying the potential stomatal regulators, including cysteine-rich secretory peptides, which require highly complex structures for their activity.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Plant Stomata/genetics , Cysteine , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Peptides/genetics , Phenotype , Gene Expression Regulation, Plant
2.
Mol Neurobiol ; 55(4): 3224-3236, 2018 Apr.
Article in English | MEDLINE | ID: mdl-28478508

ABSTRACT

Alzheimer's disease (AD) pathogenesis leads to the appearance of senile plaques due to the production and deposition of the ß-amyloid peptide (Aß). Superparamagnetic iron oxide nanoparticles (SPIONs) have potential role in the detection and imaging of Aß plaques in AD. SPIONs have shown appropriate potential in the diagnosis and treatment of AD. In the present study, the pharmacokinetics of SPIONs and its effect in the biochemical pathway of AD were analyzed using collected information. During analysis, the interaction of SPIONs with amyloid beta-42 (Aß42), a biomarker for AD progression, has been shown. Nodes represent the entities and edges represent the relation (interactions) of one node to another node. Aß42 and their interaction with other entities making up biochemical network are involved in AD mechanism in presence of SPION. The kinetic simulation was done to investigate pharmacokinetics of SPIONs for AD, where concentration was assigned of nanoparticles and other entities were applied as a kinetic irreversible simple Michaelis-Menten or mass action kinetics. Simulation was done in presence and absence of SPIONs to investigate pharmacokinetic effect in AD and explore the mechanism of Aß42 in presence of SPIONs. This study may lead to better understanding, which is required to target the metabolism of Aß42 peptide, a pivotal player in this pathology.


Subject(s)
Amyloid beta-Peptides/pharmacokinetics , Dextrans/chemistry , Magnetite Nanoparticles/chemistry , Peptide Fragments/pharmacokinetics , Signal Transduction , Systems Biology , Alzheimer Disease , Gold/chemistry , Humans , Molecular Docking Simulation
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