Addition Of Spermidine Research Articles

Decoding the influence of low temperature on biofilm development: The hidden roles of c-di-GMP

Biofilms are widely used and play important roles in biological processes. Low temperature of wastewater inhibits the development of biofilms derived from wastewater activated sludge. However, the specific mechanism of temperature on biofilm development is still unclear. This study explored the mechanism of temperature on biofilm development and found a feasible method to enhance biofilm development at low temperature. The amount of biofilm development decreased by approximately 66 % and 55 % at 4 °C and 15 °C, respectively, as compared to 28 °C. The cyclic dimeric guanosine monophosphate (c-di-GMP) concentration also decreased at low temperature and was positively correlated with extracellular polymeric substance (EPS) content, formation, and adhesion strength. Microbial community results showed that low temperature inhibited the normal survival of most microorganisms, but promoted the growth of some psychrophile bacteria like Sporosarcina, Caldilineaceae, Gemmataceae, Anaerolineaceae and Acidobacteriota. Further analysis of functional genes demonstrated that the abundance of functional genes related to the synthesis of c-di-GMP (K18968, K18967 and K13590) decreased at low temperature. Subsequently, the addition of exogenous spermidine increased the level of intracellular c-di-GMP and alleviated the inhibition effect of low temperature on biofilm development. Therefore, the possible mechanism of low temperature on biofilm development could be the inhibition of the microorganism activity and reduction of the communication level between cells, which is the closely related to the EPS content, formation, and adhesion strength. The enhancement of c-di-GMP level through the exogenous addition of spermidine provides an alternative strategy to enhance biofilm development at low temperatures. The results of this study enhance the understanding of the influence of temperature on biofilm development and provide possible strategies for enhancing biofilm development at low temperatures.

Spermidine and 1,3-Diaminopropane Have Opposite Effects on the Final Stage of Cephalosporin C Biosynthesis in High-Yielding Acremonium chrysogenum Strain.

The addition of exogenous polyamines increases the production of antibiotic cephalosporin C (CPC) in Acremonium chrysogenum high-yielding (HY) strain during fermentation on a complex medium. However, the molecular basis of this phenomenon is still unknown. In the current study, we developed a special synthetic medium on which we revealed the opposite effect of polyamines. The addition of 1,3-diaminopropane resulted in an increase in the yield of CPC by 12-15%. However, the addition of spermidine resulted in a decrease in the yield of CPC by 14-15% and accumulation of its metabolic pathway precursor, deacetylcephalosporin C (DAC); the total amount of cephems (DAC and CPC) was the same as after the addition of DAP. This indicates that spermidine, but not 1,3-diaminopropane, affects the final stage of CPC biosynthesis, associated with the acetylation of its precursor. In both cases, upregulation of biosynthetic genes from beta-lactam BGCs occurred at the same level as compared to the control; expression of transport genes was at the control level. The opposite effect may be due to the fact that N1-acetylation is much more efficient during spermidine catabolism than for 1,3-diaminopropane. The addition of spermidine, but not 1,3-diaminopropane, depleted the pool of acetyl coenzyme A by more than two-fold compared to control, which could lead to the accumulation of DAC.

Effects of Spermidine on Cell Proliferation, Migration, and Inflammatory Response in Porcine Enterocytes.

Polyamines have been demonstrated to be beneficial to porcine intestinal development. Our previous study showed that putrescine mitigates intestinal atrophy in weanling piglets and suppresses inflammatory response in porcine intestinal epithelial cells, it is still unknown the role of spermidine in mediating putrescine function. The current study aimed to investigate the effect of spermidine on the proliferation, migration, and inflammatory response in porcine intestinal epithelial cells (IPEC-J2 cell line). The effects of spermidine on proliferation and migration of IPEC-J2 cells were measured. Difluoromethyl ornithine (DFMO) and diethylglyoxal bis (guanylhydrazone) (DEGBG) were used to block the production of putrescine and spermidine, respectively. A cell inflammation model was established with lipopolysaccharides (LPS) stimulation. Gene expression and protein abundance were determined by real-time quantitative PCR and western blotting, respectively. Spermidine significantly enhanced cell proliferation in DFMO (or/and) DEGBG treated IPEC-J2 cells (p < 0.05). Pretreatment with putrescine restored cell growth inhibited by DFMO but did not prevent the decrease in cell proliferation caused by DEGBG (p > 0.05). Similarly, spermidine but not putrescine significantly elevated the rate of migration in DEGBG treated IPEC-J2 cells (p < 0.05). Spermidine deprivation by DEGBG dramatically enhanced mRNA abundance of pro-inflammatory cytokines IL-8, IL-6, and TNF-α (p < 0.05), and the addition of spermidine attenuated excessive expression of those inflammatory pro-inflammatory cytokines, moreover, spermidine but not putrescine suppressed the phosphorylation of NF-κB induced by DEGBG. Spermidine supplementation also significantly suppressed LPS-induced the expression of TNF-α. The present study highlights a novel insight that putrescine may be converted into spermidine to modulate cell proliferation, migration, and inflammatory response on porcine enterocytes.

Staphylococcus aureus 30S Ribosomal Subunit Purification and Its Biochemical and Cryo-EM Analysis.

The ribosome is a complex cellular machinery whose solved structure allowed for an incredible leap in structural biology research. Different ions bind to the ribosome, stabilizing inter-subunit interfaces and structurally linking rRNAs, proteins, and ligands. Besides cations such as K + and Mg 2+ , polyamines are known to stabilize the folding of RNA and overall structure. The bacterial ribosome is composed of a small (30S) subunit containing the decoding center and a large (50S) subunit devoted to peptide bond formation. We have previously shown that the small ribosomal subunit of Staphylococcus aureus is sensitive to changes in ionic conditions and polyamines concentration. In particular, its decoding center, where mRNA codons and tRNA anticodons interact, is prone to structural deformations in the absence of spermidine. Here, we report a detailed protocol for the purification of the intact and functional 30S, achieved through specific ionic conditions and the addition of spermidine. Using this protocol, we obtained the cryo-electron microscopy (cryo-EM) structure of the 30S-mRNA complex from S. aureus at 3.6 Å resolution. The 30S-mRNA complex formation was verified by a toeprinting assay. In this article, we also include a description of toeprinting and cryo-EM protocols. The described protocols can be further used to study the process of translation regulation. Graphical abstract.

Study on a novel buffer system to separate and quantify glycated hemoglobin A1c (HbA1c) and the mechanism between its key constituents and HbA1c

In this article, a simple capillary electrophoresis (CE) method was developed to analyze the glycated hemoglobin A1c (HbA1c) in light of the goal to monitor the long-term blood glucose concentration on diabetic patients, and it is the first time to report the binding constant of sodium tetraborate (Na2B4O7) and HbA1c in CE system. It is also found that the addition of Spermidine (Spd) can inhibit electroosmotic flow (EOF) well, thereby improving the resolution of the analytes. And a Spd concentration of a few mM is proper to provide perfect resolutions. Finally, the performance of the developed CE method was validated by analyzing actual blood specimens, of which the value of HbA1c was found to be close to the results of Sebia's CE method, showing an acceptable ability to separate hemoglobin forms. We expect this method will provide an inspiration to investigate hemoglobins (Hbs).

Spermidine Suppresses Inflammatory DC Function by Activating the FOXO3 Pathway and Counteracts Autoimmunity.

SummaryDendritic cells (DCs) function is intimately linked to microenvironment and metabolism. Type I interferons (IFNs) condition dendritic cells to respond to weak self-signals, leading to autoimmunity. However, the metabolic adaption in the process is unclear. Here, we identified spermidine as a critical metabolite impacting the metabolic fitness of DC. First, dynamic metabolome screening indicated that spermidine decreased during IFN priming and following TLR7 ligand stimulation, accompanied by metabolic change from oxidative phosphorylation to glycolysis. Second, spermidine supplement restrained the glycolysis and prevented the overactivation of IFN-α primed DC both in vivo and in vitro. Third, mechanism study uncovered that the activity of FOXO3 adapted to the metabolic change, mediating the anti-inflammatory effect of spermidine. More importantly, addition of spermidine in vivo greatly alleviated the development of psoriasis-like symptom in mice. Thus, our studies revealed metabolic changes boosting DC responses and identified spermidine as a potential therapeutic agent for autoimmune diseases.

Involvement of Spermidine in the Reduced Lifespan of Caenorhabditis elegans During Vitamin B12 Deficiency.

Vitamin B12 deficiency leads to various symptoms such as neuropathy, growth retardation, and infertility. Vitamin B12 functions as a coenzyme for two enzymes involved in amino acid metabolisms. However, there is limited information available on whether amino acid disorders caused by vitamin B12 deficiency induce such symptoms. First, free amino acid levels were determined in vitamin B12-deficient Caenorhabditis elegans to clarify the mechanisms underlying the symptoms caused by vitamin B12 deficiency. Various amino acids (valine, leucine, isoleucine, methionine, and cystathionine, among others) metabolized by vitamin B12-dependent enzymes were found to be significantly changed during conditions of B12 deficiency, which indirectly affected certain amino acids metabolized by vitamin B12-independent enzymes. For example, ornithine was significantly increased during vitamin B12 deficiency, which also significantly increased arginase activity. The accumulation of ornithine during vitamin B12 deficiency constitutes the first report. In addition, the biosynthesis of spermidine from ornithine was significantly decreased during vitamin B12 deficiency, likely due to the reduction of S-adenosylmethionine as a substrate for S-adenosylmethionine decarboxylase, which catalyzes the formation of spermidine. Moreover, vitamin B12 deficiency also demonstrated a significant reduction in worm lifespan, which was partially recovered by the addition of spermidine. Collectively, our findings suggest that decreased spermidine is one factor responsible for reduced lifespan in vitamin B12-deficient worms.

Branched-chain polyamine stabilizes RNA polymerase at elevated temperatures in hyperthermophiles.

Branched-chain polyamines (BCPAs) are unique polycations found in (hyper)thermophiles. Thermococcus kodakarensis grows optimally at 85°C and produces the BCPA N4-bis(aminopropyl)spermidine by sequential addition of decarboxylated S-adenosylmethionine (dcSAM) aminopropyl groups to spermidine (SPD) by BCPA synthase A (BpsA). The T. kodakarensisbpsA deletion mutant (DBP1) did not grow at temperatures at or above 93°C, and grew at 90°C only after a long lag period following accumulation of excess cytoplasmic SPD. This suggests that BCPA plays an essential role in cell growth at higher temperatures and raises the possibility that BCPA is involved in controlling gene expression. To examine the effects of BCPA on transcription, the RNA polymerase (RNAP) core fraction was extracted from another bpsA deletion mutant, DBP4 (RNAPDBP4), which carried a His-tagged rpoL, and its enzymatic properties were compared with those of RNAP from wild-type (WT) cells (RNAPWT). LC-MS analysis revealed that nine ribosomal proteins were detected from RNAPWT but only one form RNAPDBP4. These results suggest that BCPA increases the linkage between RNAP and ribosomes to achieve efficient coupling of transcription and translation. Both RNAPs exhibited highest transcription activity in vitro at 80°C, but the specific activity of RNAPDBP4 was lower than that of RNAPWT. Upon addition of SPD and BCPA, both increased the transcriptional activity of RNAPDBP4; however, elevation by BCPA was achieved at a tenfold lower concentration. Addition of BCPA also protected RNAPDBP4 against thermal inactivation at 90°C. These results suggest that BCPA increases transcriptional activity in T. kodakarensis by stabilizing the RNAP complex at high temperatures.

Spectroscopic and molecular docking studies on the interaction between spermidine and pancreatic elastase.

In this study, the impact of spermidine on the stability, conformation and activity of elastase was investigated at the pH of 8.5 (the optimum pH for elastase) and different temperatures (303, 313, and 323 K) using UV-vis spectrophotometry, Spectrofluorimetry, circular dichroism, and enzyme activity measurements. The empirical results were obtained and compared with those achieved by the molecular docking simulation. Spectrofluorometric results proved that with the addition of spermidine to the protein solution, the emission severity of elastase was extremely reduced. Further, the Stern-Volmer equation demonstrated that quenching was principally of the static type. Moreover, ∆H0 and ∆S0 showed a negative value, revealing that hydrogen bonds or van der Waals interactions played a critical role in the interaction between spermidine and elastase. Km and kcat [E] parameters also showed that spermidine acted as an activator for elastase. Far-UV circular dichroism also revealed that spermidine could alter the secondary structure of elastase via a partial increment within the content of the α-helix structure (from 7.8 to 8.6), while it was somewhat diminished in the β-sheet (from 29.4 to 28.8). Molecular docking simulation results also demonstrated that spermidine could bind to porcine elastase, and van der Waals forces or hydrogen bonding interactions played the main role in this binding. Spermidine, therefore, served as a partial stabilizer and an activator for the enzyme.

Putrescine biosynthesis and export genes are essential for normal growth of avian pathogenic Escherichia coli

BackgroundAvian pathogenic Escherichia coli (APEC) is the infectious agent of a wide variety of avian diseases, which causes substantial economic losses to the poultry industry worldwide. Polyamines contribute to the optimal synthesis of nucleic acids and proteins in bacteria. The objectives of this study were to investigate; i) whether APEC E. coli encodes the same systems for biosynthesis and uptake as described for E. coli K12 and ii) the role of polyamines during in vitro growth of an avian pathogenic E. coli strain (WT-ST117- O83:H4T).ResultsFollowing whole genome sequencing, polyamine biosynthesis and export genes present in E. coli MG1655 (K-12) were found to be identical in WT-ST117. Defined mutants were constructed in putrescine and spermidine biosynthesis pathways (ΔspeB, ΔspeC, ΔspeF, ΔspeB/C and ΔspeD/E), and in polyamines transport systems (ΔpotE, ΔyeeF, ΔpotABCD and ΔpotFGHI). Contrary to what was observed for MG1655, the ΔpotE-ST117 mutant was growth attenuated, regardless of putrescine supplementation. The addition of spermidine or orthinine restored the growth to the level of WT-ST117. Growth attenuation after induction of membrane stress by SDS suggested that PotE is involved in protection against this stress. The ΔspeB/C-ST117 mutant was also growth attenuated in minimal medium. The addition of putrescine or spermidine to the media restored growth rate to the wild type level. The remaining biosynthesis and transport mutants showed a growth similar to that of WT-ST117. Analysis by Ultra-High Performance Liquid Chromatography revealed that the ΔspeB/C mutant was putrescine-deficient, despite that the gene speF, which is also involved in the synthesis of putrescine, was expressed.ConclusionsDeletion of the putrescine transport system, PotE, or the putrescine biosynthesis pathway genes speB/C affected in vitro growth of APEC (ST117- O83:H4) strain, but not E. coli MG1655, despite the high similarity of the genetic make-up of biosynthesis and transport genes. Therefore, blocking these metabolic reactions may be a suitable way to prevent APEC growth in the host without disturbing the commensal E. coli population.

Plant Growth Regulators as Potential Elicitors to Increase the Contents of Phenolic Compounds and Antioxidant Capacity in Stevia Plants

Stevia rebaudiana Bertoni is mainly used as a sweetener in many countries without any reported negative effects on health. This species is also abundant in phenolic compounds, which makes this plant a good candidate to satisfy the growing demand of natural sources for sweeteners and antioxidant compounds. Exogenously applied plant growth regulators (PGRs), acting as elicitors, influence the biosynthetic pathways of secondary metabolites. In this context, methyl jasmonate (MeJa), spermidine (SPD), salicylic acid (SA), and paclobutrazol (PBZ) (100 µM) were applied to stevia plants growing in a hydroponic system to assess their effects on temporary changes (at 24, 48, and 96 h) of antioxidant capacity and phenolic compounds (total soluble phenols, flavonoids, and hydroxycinnamic acids) levels. Overall, it was observed that MeJa increased total soluble phenolic content and antioxidant activity in extracts from Stevia leaves. By contrast, the addition of SPD, SA, and PBZ did not show a significant increase in any of the evaluated parameters. The results also showed a positive and strong correlation between phenolic compounds contents and antioxidant capacity. In short, results suggest that the application of MeJa could be a feasible way to enhance the biosynthesis of high added value phytochemicals that have notable antioxidant properties, and consequently potential health benefits, in stevia plants.

Micropropagation of Kaempferia angustifolia Roscoe - An Aromatic, Essential Oil Yielding, Underutilized Medicinal Plant of Zingiberaceae Family

Kaempferia angustifolia is an aromatic, essential oil-yielding plant of the Zingiberaceae family with an ethno-medicinal repute. We standardized an effective system for micropropagation of K. angustifolia, and this is probably the very first report of in vitro culture of this species. Axillary buds were cultured on a Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of plant growth regulators (PGRs) and spermidine. Highest multiplication occurred when the MS medium was supplemented with a combination of 2.0 mg L−1 6-benzylaminopurine (BAP), 2.0 mg L−1 kinetin (KIN) and 1.0 mg L−1 α-naphthalene acetic acid (NAA). Addition of spermidine (2.0 mM) along with optimum PGRs had further improved the multiplication rate with a maximum of 6.6 ± 0.36 shoots per explant within 60 days of implantation. The number of multiplied shoots per explant increased with each subsequent regeneration cycle; and the shoots per explant increased from 6.6 ± 0.36 on the 1st regeneration cycle to 10.3 ± 0.42 on the 2nd regeneration cycle and further increased to 13.7 ± 0.37 on the 3rd regeneration cycle on the same medium composition. The best result for in vitro root induction of multiplied shoot was achieved on a half-strength MS medium fortified with 2.0 mg L−1 IBA, with a maximum of 18.5 ± 0.28 roots per shoot. Regenerated plantlets were acclimatized with 88.9 % survival rate. After 9 months of field-transfer, all these plants were harvested and rhizomes were collected. However, the present protocol can definitely be applied for large-scale propagation and commercial cultivation of K. angustifolia.

Spermidine-Induced Attraction of Like-Charged Surfaces Is Correlated with the pH-Dependent Spermidine Charge: Force Spectroscopy Characterization.

The ubiquitous molecule spermidine is known for its pivotal roles in the contact mediation, fusion, and reorganization of biological membranes and DNA. In our model system, borosilicate beads were attached to atomic force microscopy cantilevers and used to probe mica surfaces to study the details of the spermidine-induced attractions. The negative surface charges of both materials were largely constant over the measured pH range of pH 7.8 to 12. The repulsion observed between the surfaces turned into attraction after the addition of spermidine. The attractive force was correlated with the degree of spermidine protonation, which changed from +3 to +1 over the measured pH range. The force was maximal at pH 7.8. To explain the observed pH and spermidine concentration dependence, two different theoretical approaches were used: a chemical model of the charge equilibrium of spermidine and Monte-Carlo simulations of the orientation of the rodlike spermidine molecules in the gap between the borosilicate and mica surfaces. Monte-Carlo simulations of the orientational ordering of the rodlike spermidine molecules suggested the induction of attractive interactions between the surfaces if the gap was bridged by the molecules. For larger gaps, the orientational distribution function of the spermidine molecules predicted a considerable degree of parallel attachment of the molecules to the surfaces, resulting in reduced effective surface charge densities of both surfaces, which reduced their electrostatic repulsion.

Tricyclic dihydropyrimidine-based multifunctional organic nanoparticles for detection of Ag (I) ions and spermidine: Real-Time application

In the present report a dihydropyrimidine-based Biginelli compound 1 was engineered into stable organic nanoparticles via reprecipitation method. The organic nanoparticles, N1 acted as an efficient fluorescent nanosensor for selective detection of Ag (I) ions via fluorescence quenching with a detection limit of 7.9nM. Furthermore, successful detection of spermidine (an important biogenic amine) was realized through an indirect metal ion mediated approach where the quenched fluorescence of complex, C was recovered with a red-shift on addition of spermidine. The lowest detection limit for spermidine was found to be 3.2nM. The nanosensor found applicability in a wide pH range from 3 to 12, which is appropriate for evaluation in environmental and physiological conditions. The selectivity for Ag (I) ions and spermidine was further confirmed with competitive binding studies which demonstrated no interference from other analytes. Moreover, real sample analysis established the practical applicability of the developed probes. The proposed strategy provides a facile, non-invasive, sensitive and economic platform for the estimation of Ag (I) ions in environmental samples (river water) and spermidine in biological samples (urine).

Improved amplification efficiency on stool samples by addition of spermidine and its use for non-invasive detection of colorectal cancer.

BackgroundUsing quantitative methylation-specific PCR (QM-MSP) is a promising method for colorectal cancer (CRC) diagnosis from stool samples. Difficulty in eliminating PCR inhibitors of this body fluid has been extensively reported. Here, spermidine is presented as PCR facilitator for the detection of stool DNA methylation biomarkers using QM-MSP. We examined its effectiveness with NPY, PENK and WIF1, three biomarkers which we have previously shown to be of relevance to CRC.ResultsWe determined an optimal window for the amplification of the albumin (Alb) gene (100 ng of bisulfite-treated stool DNA added of 1 mM spermidine) at which we report that spermidine acts as a PCR facilitator (AE = 1680%) for SG RT-PCR. We show that the amplification of methylated PENK, NPY and WIF1 is considerably facilitated by QM-MSP as measured by an increase of CMI (Cumulative Methylation Index, i.e. the sum of the three methylation values) by a factor of 1.5 to 23 fold in individual samples, and of 10 fold in a pool of five samples.ConclusionsWe contend that spermidine greatly reduces the problems of PCR inhibition in stool samples. This observed feature, after validation on a larger sampling, could be used in the development of stool-based CRC diagnosis tests.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-015-0148-6) contains supplementary material, which is available to authorized users.

Role of Spermidine in Overwintering of Cyanobacteria.

Polyamines are found in all groups of cyanobacteria, but their role in environmental adaptation has been barely investigated. In Synechocystis sp. strain PCC 6803, inactivation of spermidine synthesis genes significantly reduced the survivability under chill (5°C)-light stress, and the survivability could be restored by addition of spermidine. To analyze the effects of spermidine on gene expression at 5°C, lacZ was expressed from the promoter of carboxy(nor)spermidine decarboxylase gene (CASDC) in Synechocystis. Synechocystis 6803::PCASDC-lacZ pretreated at 15°C showed a high level of LacZ activity for a long period of time at 5°C; without the pretreatment or with protein synthesis inhibited at 5°C, the enzyme activity gradually decreased. In a spermidine-minus mutant harboring PCASDC-lacZ, lacZ showed an expression pattern as if protein synthesis were inhibited at 5°C, even though the stability of its mRNA increased. Four other genes, including rpoA that encodes the α subunit of RNA polymerase, showed similar expression patterns. The chill-light stress led to a rapid increase of protein carbonylation in Synechocystis. The protein carbonylation then quickly returned to the background level in the wild type but continued to slowly increase in the spermidine-minus mutant. Our results indicate that spermidine promotes gene expression and replacement of damaged proteins in cyanobacteria under the chill-light stress in winter. Outbreak of cyanobacterial blooms in freshwater lakes is a worldwide environmental problem. In the annual cycle of bloom-forming cyanobacteria, overwintering is the least understood stage. Survival of Synechocystis sp. strain PCC 6803 under long-term chill (5°C)-light stress has been established as a model for molecular studies on overwintering of cyanobacteria. Here, we show that spermidine, the most common polyamine in cyanobacteria, promotes the survivability of Synechocystis under long-term chill-light stress and that the physiological function is based on its effects on gene expression and recovery from protein damage. This is the first report on the role of polyamines in survival of overwintering cyanobacteria. We also analyzed spermidine synthesis pathways in cyanobacteria on the basis of bioinformatic and experimental data.

Experimental and theoretical study of the silica particle interactions in the presence of multivalent rod-like ions.

The silica particle interactions in the presence of spermidine were systematically investigated both from experimental and theoretical points of view. The hydrodynamic radii and the corresponding polydispersity indices of the colloidal silica particles were determined by dynamic light scattering (DLS) as a function of spermidine concentration. Whereas the effective size of the silica particles increases with increasing spermidine concentration (pointing to the particle aggregation), the polydispersity index first increases reaches a maximum and then further decreases with the increasing spermidine concentration. From the mobility measurements it was concluded that the increase in spermidine concentration causes less negative values of zeta potential, meaning that the adsorption of spermidine leads to the less negative silica surface. Moreover, Monte Carlo (MC) simulations also confirmed that the addition of spermidine reduces the repulsion between silica particles. The MC concentration profiles of spermidine close to the charged silica particle are in a very good agreement with the results obtained by theory. An important motivation for our study is the effectiveness of multivalent ions to coagulate colloidal suspensions; e.g., the multivalent ions are exploited in the water purification process.

Effect of Exogenous Spermidine on the Absorption of Copper Ions in &lt;i&gt;Salix Matsudana&lt;/i&gt;

The absorption characteristics of copper and the effect of spermidine on the absorption of copper ions were investigated in Salix Matsudana in a short-term hydroponic experiment. There different concentrations of copper ions with or without the addition of spermidine in nutrition solution were conducted in this experiment. The effect of spermidine on the adsorption by S. Matsudana was also studied. 5, 10, 20 mg/L were chosen as the copper stress concentration. Absorption time and the initial copper concentration influence the absorption process in S. Matsudana. The absorption rates increased very fast at the first hours then increased slightly and attained an asymptotic value. After 72 h, the removal rates of Cu were 75.00%, 61.47% and 43.60% with the initial copper concentrations of 5, 10, 20 mg/L, respectively. The addition of spermidine influenced the absorption process of Cu by S. Matsudana. In 72 h, the treatment with spermidine increased the copper removal by S. Matsudana than the only copper treatment. Results indicated that exogenous spermidine could improve the absorption of copper in S. Matsudana and was beneficial to phytoremediation in heavy metal contaminated environment.

Polyamines Inhibit Porin-Mediated Fluoroquinolone Uptake in Mycobacteria

Polyamines decrease the permeability of the outer membrane of Escherichia coli to fluoroquinolones and β-lactams. In this study, we tested the effect of four polyamines (spermidine, spermine, cadaverine and putrescine) on fluoroquinolone uptake in Mycobacterium bovis BCG. Our results show that polyamines are also capable of reducing the permeability of the mycobacterial outer membrane to fluoroquinolones. Spermidine was most effective and demonstrated reversible dose- and pH-dependent inhibition of ciprofloxacin accumulation. The extent of this inhibition was demonstrated across the fluoroquinolone compound class to varying degrees. Furthermore, we have shown that the addition of spermidine increases the survival of M. bovis BCG after a 5-day exposure to ciprofloxacin by up to 25 times. The treatment of actively-replicating Mycobacterium tuberculosis with spermidine reduced ciprofloxacin accumulation by half while non-replicating nutrient-starved M. tuberculosis cultures lacked similar sensitivity to polyamines. Gene expression studies showed that several outer membrane proteins are significantly down–regulated during the shift to non–replication. Collectively, these characteristics of fluoroquinolone uptake in M. bovis BCG are consistent with facilitated transport by porin-like proteins and suggest that a reduction in intracellular uptake contributes to the phenotypic drug resistance demonstrated by M. tuberculosis in the non-replicating state.

Spermidine-Analogous Triamines Suppressed the Growth of &lt;i&gt;Candida albicans&lt;/i&gt;

We examined the antifungal activity of various synthetic triamines on several fungi. Among various triamines having a general structure H2N(CH2)aNH(CH2)bNH2 (a=2-5, b=3-8), some triamines (a=4 or 5) showed inhibitory effect on the growth of Candida albicans and C. tropicalis. Determination of the minimum inhibitory concentrations (MICs) of these triamines on C. albicans showed that triamine 4-8 (a=4, b=8) and triamine 5-8 had strong antifungal activity. Further analysis revealed that the antifungal effect of triamine 4-8 was fungistatic and the antifungal effect was diminished by the addition of spermidine, a physiological triamine, to the medium. These results suggested that triamine 4-8 is antagonistic to spermidine and the antifungal activity is due to the suppression of the action of intrinsic polyamines. On the agar medium, C. albicans formed microcolonies even in the presence of triamine 4-8 by long cultivation. We then observed the form of C. albicans using microscope and found that the cells cultivated with triamine 4-8 were round, similar to the yeast form, while most of the cells on the agar medium without triamine 4-8 were hyphal form. Subsequently, we investigated the synergistic effect of two compounds with triamine 4-8, cyclohexylamine and dl-α-difluoromethylornithine which are inhibitors of enzymes involving in the biosynthesis of physiological polyamines such as spermidine. The results showed that the antifungal activity of triamine 4-8 increased by the addition of these enzyme inhibitors.