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1.
Capillary electrophoresis for aluminum ion speciation: Optimized separation conditions for complex polycation mixtures.
Ouadah, N, Moire, C, Brothier, F, Kuntz, JF, Deschaume, O, Bartic, C, Cottet, H
Journal of chromatography. A. 2018;:79-86
Abstract
Aluminum chlorohydrates (ACH) are used in numerous applications and commercial products on a global scale including water treatment, catalysis or antiperspirants. They are complex mixtures of water soluble aluminum polycations of different degrees of polymerization, that are difficult to separate and quantify due to their susceptibility to depolymerize in solution when placed out of equilibrium, which is inherent to any separation process. We recently achieved the first capillary electrophoresis separation and characterization of ACH oligomers using 4-morpholineethanesulfonic acid (MES) as background electrolyte counter-ion. MES stabilizes the separated ACH oligomers during the electrophoretic process leading to highly repeatable and fast separations. In this work, the separation of ACH oligomers was further studied and perfected by varying the ionic strength, MES concentration and pH of the background electrolyte. Complex electrophoretic behavior is reported for the separation of Al13, Al30 and Na+ ions according to these experimental parameters. The transformation of the electropherograms in effective mobility scale and the use of the slope-plot approach are used to better understand the observed changes in selectivity/resolution. Optimal conditions (700 mM MES at 25 mM ionic strength containing 0.1 mM didodecyldimethylammonium bromide for dynamic capillary coating, pH 4.8) obtained for the separation of ACH oligomers are used for the baseline separation of samples difficult to analyze with other methods, including different molecular, aggregated and colloidal forms of aluminum from the Al13, Al30 and Na+ mixture, validating the rationale of the approach.
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2.
Molecules for Sensing Polyamines and Transducing Their Action in Plants.
Kusano, T, Sagor, GHM, Berberich, T
Methods in molecular biology (Clifton, N.J.). 2018;:25-35
Abstract
Polyamines play important roles in growth, development, and adaptive responses to various stresses. In the past two decades, progress in plant polyamine research has accelerated, and the key molecules and components involved in many biological events have been identified. Recently, polyamine sensors used to detect polyamine-enriched foods and polyamines derived from degrading flesh were identified in fly and zebrafish, respectively. Work has begun to identify such molecules in plants as well. Here, we summarize the current knowledge about polyamines in plants. Furthermore, we discuss the roles of key molecules, such as calcium ions, reactive oxygen species, nitric oxide, γ-aminobutyric acid, polyamine transporters, and the mitogen-activated protein kinase cascade, from the viewpoint of polyamine action.
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3.
Potential Applications of Polyamines in Agriculture and Plant Biotechnology.
Tiburcio, AF, Alcázar, R
Methods in molecular biology (Clifton, N.J.). 2018;:489-508
Abstract
The polyamines putrescine, spermidine and spermine have been implicated in a myriad of biological functions in many organisms. Research done during the last decades has accumulated a large body of evidence demonstrating that polyamines are key modulators of plant growth and development. Different experimental approaches have been employed including the measurement of endogenous polyamine levels and the activities of polyamine metabolic enzymes, the study of the effects resulting from exogenous polyamine applications and chemical or genetic manipulation of endogenous polyamine titers. This chapter reviews the role of PAs in seed germination, root development, plant architecture, in vitro plant regeneration, flowering and plant senescence. Evidence presented here indicates that polyamines should be regarded as plant growth regulators with potential applications in agriculture and plant biotechnology.
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4.
Effects of polyamines on cadmium- and copper-mediated alterations in wheat (Triticum aestivum L) and sunflower (Helianthus annuus L) seedling membrane fluidity.
Benavides, MP, Groppa, MD, Recalde, L, Verstraeten, SV
Archives of biochemistry and biophysics. 2018;:27-39
Abstract
We investigated if wheat (Wh) and sunflower (Sf) plants watering with 1 mM CdCl2 or CuCl2 for 5-15 d during germination and seedling altered membrane fluidity (MF) of their leaves and roots, and if plant pre-treatment with the polyamines (PAs) putrescine (Put), spermidine (Spd) or spermine (Spm) prevented those alterations. Cd impaired Wh and Sf growth, while Cu only affected Sf growth. Cu and Cd increased MF of leaves of both plant species, while Cd decreased MF of Sf roots. Plant treatment for 15 d with 0.1 mM Put, Spd or Spm did not affect plant growth and had opposed effects on the MF of both plants. Finally, Wh and Sf were pre-treated with PAs for either 5 or 10 days followed by metal treatment until day 15. While Put did not affect membrane MF, Spd and Spm decreased it between 5 and 10 d of plant treatment. Together, experimental results demonstrate that during plant development (a) Cd and Cu have noxious effects on plants membrane biophysical properties that could be partially responsible of their toxicity, and (b) this deleterious effect could be only partially prevented by plant pretreatment with the PAs.
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5.
Polyamine concentration is increased in thoracic ascending aorta of patients with bicuspid aortic valve.
Forte, A, Grossi, M, Bancone, C, Cipollaro, M, De Feo, M, Hellstrand, P, Persson, L, Nilsson, BO, Della Corte, A
Heart and vessels. 2018;(3):327-339
Abstract
Polyamines are cationic molecules synthesized via a highly regulated pathway, obtained from the diet or produced by the gut microbiota. They are involved in general molecular and cellular phenomena that play a role also in vascular disease. Bicuspid aortic valve (BAV) is a congenital malformation associated to a greater risk of thoracic ascending aorta (TAA) aneurysm, whose pathogenesis is not yet well understood. We focused on differential analysis of key members of polyamine pathway and on polyamine concentration in non-dilated TAA samples from patients with either stenotic tricuspid aortic valve (TAV) or BAV (diameter ≤ 45 mm), vs. normal aortas from organ donors, with the aim of revealing a potential involvement of polyamines in early aortopathy. Changes of gene expression in TAA samples were evaluated by RT-PCR. Changes of ornithine decarboxylase 1 (ODC1), a key enzyme in polyamine formation, and cationic amino acid transporter 1 (SLC7A1/CAT-1) expression were analyzed also by Western blot. ODC1 subcellular localization was assessed by immunohistochemistry. Polyamine concentration in TAA samples was evaluated by HPLC. BAV TAA samples showed an increased concentration of putrescine and spermidine vs. TAV and donor samples, together with a decreased mRNA level of polyamine anabolic enzymes and of the putative polyamine transporter SLC7A1/CAT-1. The catabolic enzyme spermidine/spermine N1-acetyltransferase 1 showed a significant mRNA increase in TAV samples only, together with a decreased concentration of spermine. The decreased expression of SLC7A1/CAT-1 and ODC1 mRNAs in BAV corresponded to increased or unchanged expression of the respective proteins. ODC was located mainly in smooth muscle cell (SMC) nucleus in TAV and donor samples, while it was present also in SMC cytoplasm in BAV samples, suggesting its activation. In conclusion, BAV, but not TAV non-dilated samples show increased polyamine concentration, accompanied by the activation of a regulatory negative feedback mechanism.
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6.
Polyamine Metabolism in Climacteric and Non-Climacteric Fruit Ripening.
Fortes, AM, Agudelo-Romero, P
Methods in molecular biology (Clifton, N.J.). 2018;:433-447
Abstract
Polyamines are small aliphatic amines that are found in both prokaryotic and eukaryotic organisms. These growth regulators have been implicated in abiotic and biotic stresses as well as plant development and morphogenesis. Several studies have also suggested a key role of polyamines during fruit set and early development. Polyamines have also been linked to fruit ripening and in the regulation of fruit quality-related traits.Recent studies indicate that during ripening of both climacteric and non-climacteric fruits, a decline in total polyamine contents is observed together with an increased catabolism of these growth regulators.In this review, we explore the current knowledge on polyamine biosynthesis and catabolism during fruit set and ripening. The study of the role of polyamine metabolism in fruit ripening indicates the possible application of these natural polycations to control ripening and postharvest decay as well as to improve fruit quality traits.
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7.
Strategies to overcome the polycation dilemma in drug delivery.
Bernkop-Schnürch, A
Advanced drug delivery reviews. 2018;:62-72
Abstract
Because of polycationic auxiliary agents such as chitosan, polyethyleneimine and cell penetrating peptides as well as cationic lipids assembling to polycationic systems, drug carriers can tightly interact with cell membranes exhibiting a high-density anionic charge. Because of these interactions the cell membrane is depolarized and becomes vulnerable to various uptake mechanisms. On their way to the target site, however, the polycationic character of all these drug carriers is eliminated by polyanionic macromolecules such as mucus glycoproteins, serum proteins, proteoglycans of the extracellular matrix (ECM) and polyanionic surface substructures of non-target cells such as red blood cells. Strategies to overcome this polycation dilemma are focusing on a pH-, redox- or enzyme-triggered charge conversion at the target site. The pH-triggered systems are making use of a slight acidic environment at the target site such as in case of solid tumors, inflammatory tissue and ischemic tissue. Due to a pH shift from 7.2 to slightly acidic mainly amino substructures of polymeric excipients are protonated or shielding groups such as 2,3 dimethylmaleic acid are cleaved off unleashing the underlying cationic character. Redox-triggered systems are utilizing disulfide linkages to bulky side chains such as PEGs masking the polycationic character. Under mild reducing conditions such as in the tumor microenvironment these disulfide bonds are cleaved. Enzyme-triggered systems are targeting enzymes such as alkaline phosphatase, matrix metalloproteinases or hyaluronidase in order to eliminate anionic moieties via enzymatic cleavage resulting in a charge conversion from negative to positive. Within this review an overview about the pros and cons of these systems is provided.
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8.
Characterization of G-Quadruplexes in Chlamydomonas reinhardtii and the Effects of Polyamine and Magnesium Cations on Structure and Stability.
Vinyard, WA, Fleming, AM, Ma, J, Burrows, CJ
Biochemistry. 2018;(47):6551-6561
Abstract
Chlamydomonas reinhardtii is a green alga with a very GC-rich genome (67%) and a high density of potential G-quadruplex-forming sequences (PQSs). Using the Ensembl Plants DNA database, 19 PQSs were selected, and their ability to fold in vitro was examined using four experimental methods. Our results support in vitro folding of 18 of the 19 PQSs selected for study. The high physiological polyamine concentrations in C. reinhardtii create unique conditions for studying G4 folding. We investigated whether high polyamine concentrations affect the stability and structural fold of two polymorphic G4s selected from the cohort of PQSs. The two polymorphic G4s selected were found to be greatly stabilized when studied at the physiologically high polyamine concentrations. Lastly, the effects of physiologically relevant Mg2+ concentrations were tested on both of the polymorphic G4s, and one of the G4s shifted from a dynamic mixture of folds to favor a parallel fold in the presence of Mg2+. Our work supports the concept of folding of G4s under the unique conditions observed in C. reinhardtii, and these structures, being located in promoter regions of DNA repair and photosynthetic genes, might be relevant structures in the physiology of C. reinhardtii.
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9.
The Arabidopsis N-Acetylornithine Deacetylase Controls Ornithine Biosynthesis via a Linear Pathway with Downstream Effects on Polyamine Levels.
Molesini, B, Zanzoni, S, Mennella, G, Francese, G, Losa, A, L Rotino, G, Pandolfini, T
Plant & cell physiology. 2017;(1):130-144
Abstract
Arabidopsis thaliana At4g17830 codes for a protein showing sequence similarity with the Escherichia coli N-acetylornithine deacetylase (EcArgE), an enzyme implicated in the linear ornithine (Orn) biosynthetic pathway. In plants, N-acetylornithine deacetylase (NAOD) activity has yet to be demonstrated; however, At4g17830-silenced and mutant (atnaod) plants display an impaired reproductive phenotype and altered foliar levels of Orn and polyamines (PAs). Here, we showed the direct connection between At4g17830 function and Orn biosynthesis, demonstrating biochemically that At4g17830 codes for a NAOD. These results are the first experimental proof that Orn can be produced in Arabidopsis via a linear pathway. In this study, to identify the role of AtNAOD in reproductive organs, we carried out a transcriptomic analysis on atnaod mutant and wild-type flowers. In the atnaod mutant, the most relevant effects were the reduced expression of cysteine-rich peptide-coding genes, known to regulate male-female cross-talk during reproduction, and variation in the expression of genes involved in nitrogen:carbon (N:C) status. The atnaod mutant also exhibited increased levels of sucrose and altered sensitivity to glucose. We hypothesize that AtNAOD participates in Orn and PA homeostasis, contributing to maintain an optimal N:C balance during reproductive development.
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10.
Modulatory Effects of Exogenously Applied Polyamines on Postharvest Physiology, Antioxidant System and Shelf Life of Fruits: A Review.
Sharma, S, Pareek, S, Sagar, NA, Valero, D, Serrano, M
International journal of molecular sciences. 2017;(8)
Abstract
Polyamines (PAs) are natural compounds involved in many growth and developmental processes in plants, and, specifically in fruits, play a vital role regulating its development, ripening and senescence processes. Putrescine (PUT), spermine (SPE), and spermidine (SPD) are prominent PAs applied exogenously to extend shelf life of fruits. They also originate endogenously during developmental phases of horticultural crops and simultaneously affect the quality attributes and shelf life. Their anti-ethylene nature is being exploited to enhance the shelf life when exogenously applied on fruits. In growth and development of fruits, PA levels generally fall, which marks the beginning of senescence at postharvest phase. PUT, SPE and SPD treatments are being applied during postharvest phase to prolong the shelf life. They enhance the shelf life of fruits by reducing respiration rate, ethylene release and enhance firmness and quality attributes in fruits. PAs have a mitigating impact on biotic and abiotic stresses including chilling injury (CI) in tropical and sub-tropical fruits. PAs are environment friendly in nature and are biodegradable without showing any negative effect on environment. Biotechnological interventions by using chimeric gene constructs of PA encoding genes has boosted the research to develop transgenic fruits and vegetables which would possess inherent or in situ mechanism of enhanced biosynthesis of PAs at different stages of development and thereby will enhance the shelf life and quality in fruits. Internal and external quality attributes of fruits are improved by modulation of antioxidant system and by strengthening biophysical morphology of fruits by electrostatic interaction between PAs and phospholipids in the cell wall.