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Nutrients Apr 2023(1) Background: Spermidine is a biogenic polyamine that plays a crucial role in mammalian metabolism. As spermidine levels decline with age, spermidine supplementation... (Randomized Controlled Trial)
Randomized Controlled Trial
High-Dose Spermidine Supplementation Does Not Increase Spermidine Levels in Blood Plasma and Saliva of Healthy Adults: A Randomized Placebo-Controlled Pharmacokinetic and Metabolomic Study.
(1) Background: Spermidine is a biogenic polyamine that plays a crucial role in mammalian metabolism. As spermidine levels decline with age, spermidine supplementation is suggested to prevent or delay age-related diseases. However, valid pharmacokinetic data regarding spermidine remains lacking. Therefore, for the first time, the present study investigated the pharmacokinetics of oral spermidine supplementation. (2) Methods: This study was designed as a randomized, placebo-controlled, triple-blinded, two-armed crossover trial with two 5-day intervention phases separated by a washout phase of 9 days. In 12 healthy volunteers, 15 mg/d of spermidine was administered orally, and blood and saliva samples were taken. Spermidine, spermine, and putrescine were quantified by liquid chromatography-mass spectrometry (LC-MS/MS). The plasma metabolome was investigated using nuclear magnetic resonance (NMR) metabolomics. (3) Results: Compared with a placebo, spermidine supplementation significantly increased spermine levels in the plasma, but it did not affect spermidine or putrescine levels. No effect on salivary polyamine concentrations was observed. (4) Conclusions: This study's results suggest that dietary spermidine is presystemically converted into spermine, which then enters systemic circulation. Presumably, the in vitro and clinical effects of spermidine are at least in part attributable to its metabolite, spermine. It is rather unlikely that spermidine supplements with doses <15 mg/d exert any short-term effects.
Topics: Animals; Adult; Humans; Spermidine; Spermine; Putrescine; Chromatography, Liquid; Saliva; Tandem Mass Spectrometry; Polyamines; Plasma; Dietary Supplements; Mammals
PubMed: 37111071
DOI: 10.3390/nu15081852 -
Biomolecules Apr 2021Astrocytes serve essential roles in human brain function and diseases. Growing evidence indicates that astrocytes are central players of the feedback modulation of...
Astrocytes serve essential roles in human brain function and diseases. Growing evidence indicates that astrocytes are central players of the feedback modulation of excitatory Glu signalling during epileptiform activity via Glu-GABA exchange. The underlying mechanism results in the increase of tonic inhibition by reverse operation of the astroglial GABA transporter, induced by Glu-Na symport. GABA, released from astrocytes, is synthesized from the polyamine (PA) putrescine and this process involves copper amino oxidase. Through this pathway, putrescine can be considered as an important source of inhibitory signaling that counterbalances epileptic discharges. Putrescine, however, is also a precursor for spermine that is known to enhance gap junction channel communication and, consequently, supports long-range Ca signaling and contributes to spreading of excitatory activity through the astrocytic syncytium. Recently, we presented the possibility of neuron-glia redox coupling through copper (Cu/Cu) signaling and oxidative putrescine catabolism. In the current work, we explore whether the Cu/Cu homeostasis is involved in astrocytic control on neuronal excitability by regulating PA catabolism. We provide supporting experimental data underlying this hypothesis. We show that the blockade of copper transporter (CTR1) by AgNO (3.6 µM) prevents GABA transporter-mediated tonic inhibitory currents, indicating causal relationship between copper (Cu/Cu) uptake and the catabolism of putrescine to GABA in astrocytes. In addition, we show that MnCl (20 μM), an inhibitor of the divalent metal transporter DMT1, also prevents the astrocytic Glu-GABA exchange. Furthermore, we observed that facilitation of copper uptake by added CuCl (2 µM) boosts tonic inhibitory currents. These findings corroborate the hypothesis that modulation of neuron-glia coupling by copper uptake drives putrescine → GABA transformation, which leads to subsequent Glu-GABA exchange and tonic inhibition. Findings may in turn highlight the potential role of copper signaling in fine-tuning the activity of the tripartite synapse.
Topics: Animals; Astrocytes; Cells, Cultured; Copper; Copper Transport Proteins; GABA Plasma Membrane Transport Proteins; Inhibitory Postsynaptic Potentials; Mice; Neurons; Putrescine; gamma-Aminobutyric Acid
PubMed: 33921742
DOI: 10.3390/biom11040604 -
The Journal of Biological Chemistry 2021Polyamines, such as putrescine, spermidine, and spermine, are physiologically important polycations, but the transporters responsible for their uptake in mammalian cells...
Polyamines, such as putrescine, spermidine, and spermine, are physiologically important polycations, but the transporters responsible for their uptake in mammalian cells remain poorly characterized. Here, we reveal a new component of the mammalian polyamine transport system using CHO-MG cells, a widely used model to study alternative polyamine uptake routes and characterize polyamine transport inhibitors for therapy. CHO-MG cells present polyamine uptake deficiency and resistance to a toxic polyamine biosynthesis inhibitor methylglyoxal bis-(guanylhydrazone) (MGBG), but the molecular defects responsible for these cellular characteristics remain unknown. By genome sequencing of CHO-MG cells, we identified mutations in an unexplored gene, ATP13A3, and found disturbed mRNA and protein expression. ATP13A3 encodes for an orphan P5B-ATPase (ATP13A3), a P-type transport ATPase that represents a candidate polyamine transporter. Interestingly, ATP13A3 complemented the putrescine transport deficiency and MGBG resistance of CHO-MG cells, whereas its knockdown in WT cells induced a CHO-MG phenotype demonstrated as a decrease in putrescine uptake and MGBG sensitivity. Taken together, our findings identify ATP13A3, which has been previously genetically linked with pulmonary arterial hypertension, as a major component of the mammalian polyamine transport system that confers sensitivity to MGBG.
Topics: Adenosine Triphosphatases; Animals; Biological Transport; CHO Cells; Cricetinae; Cricetulus; Enzyme Inhibitors; Membrane Transport Proteins; Mitoguazone; Mutation; Polyamines; Putrescine; Whole Genome Sequencing
PubMed: 33310703
DOI: 10.1074/jbc.RA120.013908 -
Biosensors Nov 2022This work presents a multipurpose and multilayered stainless steel microneedle sensor for the in situ redox potential monitoring in food and drink samples, termed MN...
This work presents a multipurpose and multilayered stainless steel microneedle sensor for the in situ redox potential monitoring in food and drink samples, termed MN redox sensor. The MN redox sensor was fabricated by layer-by-layer (LbL) approach. The in-tube multilayer coating comprised carbon nanotubes (CNTs)/cellulose nanocrystals (CNCs) as the first layer, polyaniline (PANI) as the second layer, and the ferrocyanide redox couple as the third layer. Using cyclic voltammetry (CV) as a transduction method, the MN redox sensor showed facile electron transfer for probing both electrical capacitance and redox potential, useful for both analyte specific and bulk quantification of redox species in various food and drink samples. The bulk redox species were quantified based on the anodic/cathodic redox peak shifts (E/E) on the voltammograms resulting from the presence of redox-active species. The MN redox sensor was applied to detect selected redox species including ascorbic acid, HO, and putrescine, with capacitive limits of detection (LOD) of 49.9, 17.8, and 263 ng/mL for each species, respectively. For the bulk determination of redox species, the MN redox sensor displayed LOD of 5.27 × 10, 55.4, and 25.8 ng/mL in ascorbic acid, HO, and putrescine equivalents, respectively. The sensor exhibited reproducibility of ~ 1.8% relative standard deviation (%RSD). The MN redox sensor was successfully employed for the detection of fish spoilage and antioxidant quantification in king mushroom and brewed coffee samples, thereby justifying its potential for food quality and food safety applications. Lastly, the portability, reusability, rapid sampling time, and capability of in situ analysis of food and drink samples makes it amenable for real-time sensing applications.
Topics: Animals; Nanotubes, Carbon; Electrochemical Techniques; Food Analysis; Hydrogen Peroxide; Putrescine; Reproducibility of Results; Ascorbic Acid
PubMed: 36354510
DOI: 10.3390/bios12111001 -
Plant, Cell & Environment Nov 2020Polyamines are small amines that accumulate during stress and contribute to disease resistance through as yet unknown signaling pathways. Using a comprehensive...
Polyamines are small amines that accumulate during stress and contribute to disease resistance through as yet unknown signaling pathways. Using a comprehensive RNA-sequencing analysis, we show that early transcriptional responses triggered by each of the most abundant polyamines (putrescine, spermidine, spermine, thermospermine and cadaverine) exhibit specific quantitative differences, suggesting that polyamines (rather than downstream metabolites) elicit defense responses. Signaling by putrescine, which accumulates in response to bacteria that trigger effector triggered immunity (ETI) and systemic acquired resistance (SAR), is largely dependent on the accumulation of hydrogen peroxide, and is partly dependent on salicylic acid (SA), the expression of ENHANCED DISEASE SUSCEPTIBILITY (EDS1) and NONEXPRESSOR of PR GENES1 (NPR1). Putrescine elicits local SA accumulation as well as local and systemic transcriptional reprogramming that overlaps with SAR. Loss-of-function mutations in arginine decarboxylase 2 (ADC2), which is required for putrescine synthesis and copper amine oxidase (CuAO), which is involved in putrescine oxidation, compromise basal defenses, as well as putrescine and pathogen-triggered systemic resistance. These findings confirm that putrescine elicits ROS-dependent SA pathways in the activation of plant defenses.
Topics: Arabidopsis; Cadaverine; Gene Expression Profiling; Plant Growth Regulators; Plant Leaves; Putrescine; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Salicylic Acid; Signal Transduction; Spermidine; Spermine
PubMed: 32839979
DOI: 10.1111/pce.13874 -
Scientific Reports Mar 2022Aromatic rice (Oryza sativa) fetches a premium price due to the pleasant aroma. The major aroma compound 2-acetyl-1-pyrroline (2AP) has been found to be enhanced under...
Aromatic rice (Oryza sativa) fetches a premium price due to the pleasant aroma. The major aroma compound 2-acetyl-1-pyrroline (2AP) has been found to be enhanced under stress. This condition can be considered to study the genes, precursors, enzymes, and metabolites involved in elevated levels of 2AP biosynthesis. In the present study, 100 mM salt treatment was given to two aromatic rice cultivars Ambemohar-157 (A-157) and Basmati-370 (B-370) at the vegetative stage (VS). After salt treatment, in the leaves, 2AP contents were elevated by 2.2 and 1.8 fold in A-157 and B-370, respectively. Under these elevated 2AP conditions, the precursor amino acids (glutamate, putrescine, ornithine, and proline), their related genes, enzymes, and metabolites (methylglyoxal and γ-aminobutyric acid (GABA) related to 2AP biosynthesis were analyzed. In addition, agronomic characters were also studied. It was observed that the proline content was enhanced in both the cultivars by 29% (A-157) and 40% (B-370) as compared to control. The Δ-pyrroline-5-carboxylate synthetase (P5CS) enzyme activity was increased in salt-treated plants leaf tissue by 31% (A-157) and 40% (B-370) compared to control. The P5CS gene expression was enhanced by A-157 (1.8 fold) and B-370 (2.2 fold) compared to control, putrescine content in A-157 and B-370 decreased by 2.5 and 2.7 fold respectively as compared to control. The ornithine decarboxylase (ODC) activity was enhanced in A-157 (12%) and B-370 (35%) over control. Further, ODC gene expression was enhanced in both the cultivars A-157 (1.5 fold) and B-370 (1.3 fold). The diamino oxidase (DAO) enzyme activity was increased by 28% (A-157) and 35% (B-370) respectively over control. The GABA content marginally increased over control in both the cultivars namely, A-157 (1.9%) and B-370 (9.5%). The methylglyoxal levels were enhanced by 1.4 fold in A-157 and 1.6 fold in B-370. Interestingly, the enhancement in 2AP in the vegetative stage also helped to accumulate it in mature grains (twofold in A-157 and 1.5 fold in B-370) without test weight penalty. The study indicated that the ornithine and proline together along with methylglyoxal contribute towards the enhancement of 2AP under salt stress.
Topics: Amino Acids; Ornithine; Oryza; Proline; Putrescine; Pyrroles; Pyruvaldehyde; Salt Stress; gamma-Aminobutyric Acid
PubMed: 35273240
DOI: 10.1038/s41598-022-07844-7 -
BMC Plant Biology Mar 2023Cold stress is an effective factor in reducing production and injuring fruit trees. Various materials, such as salicylic acid, ascorbic acid, and putrescine, are used to...
BACKGROUND
Cold stress is an effective factor in reducing production and injuring fruit trees. Various materials, such as salicylic acid, ascorbic acid, and putrescine, are used to alleviate the damage of abiotic stress.
RESULTS
The effect of different treatments of putrescine, salicylic acid, and ascorbic acid on alleviating the damage of frost stress (- 3 °C) to grapes 'Giziluzum' was investigated. Frost stress increased the amount of HO, MDA, proline, and MSI. On the other hand, it decreased the concentration of chlorophyll and carotenoids in the leaves. Putrescine, salicylic acid and ascorbic acid significantly increased the activities of catalase, guaiacol peroxidase, ascorbate peroxidase, and superoxide dismutase under frost stress. Following frost stress, the grapes treated with putrescine, salicylic acid, and ascorbic acid showed higher levels of DHA, AsA, and AsA/DHA than the untreated grapes. Our results showed that the treatment with ascorbic acid outperformed the other treatments in adjusting frost stress damages.
CONCLUSION
The use of compounds, such as ascorbic ac id, salicylic acid, and putrescine, modulates the effects of frost stress, thereby increasing the antioxidant defense system of cells, reducing its damage, and stabilizing stable cell conditions, so it can be used to reduce frost damage to different grape cultivars.
Topics: Ascorbic Acid; Vitis; Salicylic Acid; Putrescine; Hydrogen Peroxide; Antioxidants
PubMed: 36899321
DOI: 10.1186/s12870-023-04126-w -
International Journal of Molecular... Nov 2022An extreme thermophile, , produces 16 different polyamines including long-chain and branched-chain polyamines. The composition and content of polyamines in the...
An extreme thermophile, , produces 16 different polyamines including long-chain and branched-chain polyamines. The composition and content of polyamines in the thermophile cells change not only with growth temperature but also with pH changes. In particular, cell growth decreased greatly at alkaline medium together with significant changes in the composition and content of polyamines. The amounts of tetraamines (spermine and its homologs) markedly decreased at alkaline pH. Thus, we knocked out the gene, which is involved in the biosynthesis of tetraamines, and changes of composition of polyamines with pH changes in the mutant cells were studied. Cell growth in the Δ strain was decreased compared with that of the wild-type strain for all pHs, suggesting that tetraamines are important for cell proliferation. Interestingly, the amount of spermidine decreased and that of putrescine increased in wild-type cells at elevated pH, although lacks a putrescine synthesizing pathway. In addition, polyamines possessing a diaminobutane moiety, such as spermine, decreased greatly at high pH. We assessed whether the gene encoding aminopropylagmatine ureohydrolase (TtSpeB) is directly involved in the synthesis of putrescine. The catalytic assay of the purified enzyme indicated that TtSpeB accepts agmatine as its substrate and produces putrescine due to the change in substrate specificity at high pH. These results suggest that pH stress was exacerbated upon intracellular depletion of polyamines possessing a diaminobutane moiety induced by unusual changes in polyamine biosynthesis under high pH conditions.
Topics: Thermus thermophilus; Spermine; Putrescine; Polyamines; Spermidine
PubMed: 36362306
DOI: 10.3390/ijms232113523 -
Scientific Reports Oct 2022Fish use odor to avoid exposure to predation and disease. Harnessing these odors as repellents is proving useful for management initiatives that conserve native species...
Fish use odor to avoid exposure to predation and disease. Harnessing these odors as repellents is proving useful for management initiatives that conserve native species or control invasive populations. Here, we evaluated the behavioral response of invasive sea lamprey to putrescine, a decay molecule that many prey organisms avoid. Putrescine is found in tissue extracts that contain sea lamprey alarm cue, and human saliva, two mixtures known to elicit flight and avoidance responses in migratory sea lamprey. We used two behavioral assays to evaluate metrics of repellency: behavioral preference (space use) and change in activity rates and found context-dependent results. In smaller assays with individual fish, we found that putrescine had no effect on sea lamprey activity but did induce avoidance. In larger assays with multiple animals, sea lamprey did not avoid putrescine. Our results also showed consistent changes in activity and avoidance behavior in sea lamprey exposed to alarm cue in the smaller assay, concluding that this design could prove useful as a high-throughput screening tool. We also investigated a novel odor identified in sea lamprey skin, petromyzonacil, and found no behavioral effects to this odor on its own or in synergy with putrescine. Our results show limited evidence that putrescine acts as robust repellent for sea lamprey and highlight the importance of environmental context when interpreting avoidance behavior in laboratory settings.
Topics: Animals; Humans; Odorants; Petromyzon; Predatory Behavior; Putrescine; Tissue Extracts
PubMed: 36253421
DOI: 10.1038/s41598-022-22143-x -
Medical Sciences (Basel, Switzerland) Aug 2022The major intracellular polyamines spermine and spermidine are abundant and ubiquitous compounds that are essential for cellular growth and development. Spermine...
The major intracellular polyamines spermine and spermidine are abundant and ubiquitous compounds that are essential for cellular growth and development. Spermine catabolism is mediated by spermine oxidase (SMOX), a highly inducible flavin-dependent amine oxidase that is upregulated during excitotoxic, ischemic, and inflammatory states. In addition to the loss of radical scavenging capabilities associated with spermine depletion, the catabolism of spermine by SMOX results in the production of toxic byproducts, including HO and acrolein, a highly toxic aldehyde with the ability to form adducts with DNA and inactivate vital cellular proteins. Despite extensive evidence implicating SMOX as a key enzyme contributing to secondary injury associated with multiple pathologic states, the lack of potent and selective inhibitors has significantly impeded the investigation of SMOX as a therapeutic target. In this study, we used a virtual and physical screening approach to identify and characterize a series of hit compounds with inhibitory activity against SMOX. We now report the discovery of potent and highly selective SMOX inhibitors (IC 0.54 μM, Ki 1.60 μM) and (IC 0.23 μM, K 0.46 μM), which are the most potent SMOX inhibitors reported to date. We hypothesize that these selective SMOX inhibitors will be useful as chemical probes to further elucidate the impact of polyamine catabolism on mechanisms of cellular injury.
Topics: Acrolein; Flavins; Hydrogen Peroxide; Oxidoreductases Acting on CH-NH Group Donors; Polyamines; Spermidine; Spermine; Polyamine Oxidase
PubMed: 36135832
DOI: 10.3390/medsci10030047