RESUMEN

Memory consolidation is associated with the regulation of protein kinases, which impact synaptic functions and promote synaptogenesis. The administration of spermidine (SPD) has been shown to modulate major protein kinases associated with memory improvement, including the Ca2+-dependent protein kinase (PKC) and cAMP-dependent protein kinase (PKA), key players in the cAMP response element-binding protein (CREB) activation. Nevertheless, the initial mechanism underlying SPD-mediated memory consolidation remains unknown, as we hypothesize a potential involvement of the memory consolidation precursor, Ca2+/calmodulin-dependent protein kinase II-α (CaMKIIα), in this process. Based on this, our study aimed to investigate potential interactions among PKC, PKA, and CREB activation, mediated by CaMKIIα activation, in order to elucidate the SPD memory consolidation pathway. Our findings suggest that the post-training administration of the CaMKII inhibitor, KN-62 (0.25 nmol, intrahippocampal), prevented the memory enhancement induced by SPD (0.2 nmol, intrahippocampal) in the inhibitory avoidance task. Through western immunoblotting, we observed that phosphorylation of CaMKIIα in the hippocampus was facilitated 15 min after intrahippocampal SPD administration, resulting in the activation of PKA and CREB, 180 min after infusion, suggesting a possible sequential mechanism, since SPD with KN-62 infusion leads to a downregulation in CaMKIIα/PKA/CREB pathway. However, KN-62 does not alter the memory-facilitating effect of SPD on PKC, possibly demonstrating a parallel cascade in memory acquisition via PKA, without modulating CAMKIIα. These results suggest that memory enhancement induced by SPD administration involves crosstalk between CaMKIIα and PKA/CREB, with no PKC interaction.

Asunto(s)

Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina , Proteína de Unión a Elemento de Respuesta al AMP Cíclico , Proteínas Quinasas Dependientes de AMP Cíclico , Memoria , Ratas Wistar , Transducción de Señal , Espermidina , Animales , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Ratas , Espermidina/farmacología , Masculino , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Memoria/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Fosforilación/efectos de los fármacos , Sulfonamidas/farmacología , Bencilaminas/farmacología , Bencilaminas/administración & dosificación , Reacción de Prevención/efectos de los fármacos , Proteína Quinasa C/metabolismo , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/análogos & derivados

RESUMEN

Marolo (Annona crassiflora) is an underutilized Brazilian Cerrado fruit with few reports in the literature about its bioactive compounds and functional properties. In this context, the chemoprevention against the carcinogen 1,2-dimethylhydrazine (DMH)-induced pre-neoplastic lesions in Wistar rat colon was investigated and correlated with marolo's antioxidant activity and the contents of phenolic compounds and bioactive amines. Total phenolic compounds (TPC) and total flavonoids compounds (TFC) were determined in the marolo pulp extract by spectrophotometric and Ultra-Performance Liquid Chromatography and diode array detection (UPLC-DAD) analysis. Free bioactive amines were determined by High Performance Liquid Chromatography and fluorescence detection (HPLC-FLD) after post column derivatization with o-phthalaldehyde. In addition, the in vitro antioxidant activity was determined by DPPH, and ABTS. Wistar rats were treated orally with marolo pulp at 0.7, 1.4 and 2.8 g/kg body weight (bw)/day added to a standard ration. Four subcutaneous injections of DMH (40 mg/kg bw) were used to induce a pre-neoplastic lesion that was assessed by the aberrant crypt foci (ACF) assay. The marolo pulp (fresh weigh) showed high content of total phenolic compounds (9.16 mg GAE/g), with predominance of chlorogenic acid (1.86 µg/g) and epicatechin (0.99 µg/g), and total flavonoids (7.26 mg CE/g), ∼85 % of the TPC. The marolo pulp had significant contents of tyramine (31.97 mg/kg), putrescine (20.65 mg/kg), and spermidine (6.32 mg/kg). The marolo pulp inhibited (p < 0.05) pre-neoplastic lesions induced by DMH administration at the all concentrations tested. These findings indicate that marolo pulp has a colon carcinogenesis chemopreventive effect, which could be due to, at least in parts, its antioxidant action associated with its phenolics and flavonoids content as well of spermidine.

Asunto(s)

Antioxidantes , Fenol , Ratas , Animales , Antioxidantes/farmacología , Antioxidantes/análisis , Ratas Wistar , Espermidina , 1,2-Dimetilhidrazina/toxicidad , Carcinogénesis , Fenoles/farmacología , Fenoles/análisis , Flavonoides/farmacología

RESUMEN

Polyamines (Pas) are short molecules that exhibit two or three amine groups that are positively charged at a physiological pH. These small molecules are present in high concentrations in a wide variety of organisms and tissues, suggesting that they play an important role in cellular physiology. Polyamines include spermine, spermidine, and putrescine, which play important roles in age-related diseases that have not been completely elucidated. Aging is a natural process, defined as the time-related deterioration of the physiological functions; it is considered a risk factor for degenerative diseases such as cardiovascular, neurodegenerative, and musculoskeletal diseases; arthritis; and even cancer. In this review, we provide a new perspective on the participation of Pas in the cellular and molecular processes related to age-related diseases, focusing our attention on important degenerative diseases such as Alzheimerߣs disease, Parkinsonߣs disease, osteoarthritis, sarcopenia, and osteoporosis. This new perspective leads us to propose that Pas function as novel biomarkers for age-related diseases, with the main purpose of achieving new molecular alternatives for healthier aging.

Asunto(s)

Poliaminas , Espermidina , Espermina/fisiología , Putrescina

RESUMEN

The influence of under-fermented (UF) cocoa (0 to 65 %) on bioactive amines in chocolate and their in vitro bioaccessibility was investigated. The same amines were found in all treatments; however, treatments were divided into two groups regarding total amines [0 & 20 % UF (34 mg/kg) and 35 to 65 % UF (17 mg/kg)] and phenolic levels [lower and higher, respectively]. Serotonin, tyramine, putrescine, cadaverine, agmatine and phenylethylamine were higher in chocolate with ≤ 20 % UF cocoa. Histamine and spermidine were not affected. Digestibility studies indicated that low levels of amines were present in the oral phase. Gastric digestion was effective in releasing tyramine, spermidine and phenylethylamine from conjugates. Serotonin and agmatine were not detected after in vitro digestion of chocolate with ≥ 35 % UF cocoa. Histamine was released during in vitro intestinal digestion. By adding different proportions of UF cocoa during chocolate production, the levels and bioaccessibility of amines can be modulated.

Asunto(s)

Agmatina , Cacao , Chocolate , Espermidina , Histamina , Serotonina , Tiramina , Fenetilaminas , Fermentación , Aminas Biogénicas

RESUMEN

To succeed in plant invasion, phytopathogenic bacteria rely on virulence mechanisms to subvert plant immunity and create favorable conditions for growth. This process requires a precise regulation in the production of important proteins and metabolites. Among them, the family of compounds known as polyamines have attracted considerable attention as they are involved in important cellular processes, but it is not known yet how phytopathogenic bacteria regulate polyamine homeostasis in the plant environment. In the present study, we performed a meta-analysis of publicly available transcriptomic data from experiments conducted on bacteria to begin delving into this topic and better understand the regulation of polyamine metabolism and its links to pathogenicity. We focused our research on Pseudomonas syringae, an important phytopathogen that causes disease in many economically valuable plant species. Our analysis discovered that polyamine synthesis, as well as general gene expression activation and energy production are induced in the early stages of the disease. On the contrary, synthesis of these compounds is inhibited whereas its transport is upregulated later in the process, which correlates with the induction of virulence genes and the metabolism of nitrogen and carboxylic acids. We also found that activation of plant defense mechanisms affects bacterial polyamine synthesis to some extent, which could reduce bacterial cell fitness in the plant environment. Furthermore, data suggest that a proper bacterial response to oxidative conditions requires a decrease in polyamine production. The implications of these findings are discussed.

RESUMEN

ATP-Binding Cassette transporters (ABC transporters) are protein complexes involved in the import and export of different molecules, including ions, sugars, peptides, drugs, and others. Due to the diversity of substrates, they have large relevance in physiological processes such as virulence, pathogenesis, and antimicrobial resistance. In Xanthomonas citri subsp. citri, the phytopathogen responsible for the citrus canker disease, 20% of ABC transporters components are expressed under infection conditions, including the putative putrescine/polyamine ABC transporter, PotFGHI. Polyamines are ubiquitous molecules that mediate cell growth and proliferation and play important role in bacterial infections. In this work, we characterized the X. citri periplasmic-binding protein PotF (XAC2476) using bioinformatics, biophysical and structural methods. PotF is highly conserved in Xanthomonas sp. genus, and we showed it is part of a set of proteins related to the import and assimilation of polyamines in X. citri. The interaction of PotF with putrescine and spermidine was direct and indirectly shown through fluorescence spectroscopy analyses, and experiments of circular dichroism (CD) and small-angle X-ray scattering (SAXS), respectively. The protein showed higher affinity for spermidine than putrescine, but both ligands induced structural changes that coincided with the closing of the domains and increasing of thermal stability.

RESUMEN

Tetragonisca angustula honey was fractioned in a SiO2 column to furnish three fractions (A-C) in which four hydroxycinnamic acid-Spermidine amides (HCAAs), known as N', N″, N‴-tris-p-coumaroyl spermidine, N', N″-dicaffeoyl, N‴-coumaroyl spermidine, N', N″, N‴-tris-caffeoyl spermidine and N', N″-dicaffeoyl and N‴-feruloyl spermidine were identified in the fractions B and C by electrospray ionization tandem mass spectrometry. A primary culture model previously infected with Neospora caninum (72 h) was used to evaluate the honey fractions (A-C) for two-time intervals: 24 and 72 h. Parasitic reduction ranged from 38% on fraction C (12.5 µg/ml), after 24 h, to 54% and 41% with fractions B and C (25 µg/ml) after 72 h of treatment, respectively. Additionally, HCAAs did not show any cell toxicity for 24 and 72 h. For infected cultures (72 h), the active fractions B (12.5 µg/ml) and C (25 µg/ml) decreased their NO content. In silico studies suggest that HCAAs may affect the parasite's redox pathway and improve the oxidative effect of NO released from infected cells. Here, we presented for the first time, that HCAAs from T. angustula honey have the potential to inhibit the growth of N. caninum protozoa.

Asunto(s)

Antiprotozoarios/farmacología , Abejas , Miel , Neospora/efectos de los fármacos , Espermidina/química , Amidas/química , Animales , Antiprotozoarios/química , Brasil , Células Cultivadas , Coccidiosis/tratamiento farmacológico , Simulación por Computador , Ácidos Cumáricos/química , NADH NADPH Oxidorreductasas/antagonistas & inhibidores , Neuroglía/efectos de los fármacos , Neuroglía/parasitología , Óxido Nítrico/metabolismo , Ratas Wistar , Espermidina/análisis

RESUMEN

The polyamine content of human breast milk, which is the first exogenous source of polyamines for the newborn, can be affected by several factors associated with the mother, the infant, or breastfeeding itself. The aim of this study was to evaluate the influence of different breastfeeding factors on the polyamines found in human milk. For this study, a cohort of 83 mothers was considered for up to 4 months, and a subgroup of 33 mothers were followed during the first six months of breastfeeding. Two breast milk samples were collected at each sampling point (foremilk and hindmilk) and the polyamine content was determined by UHPLC-FL. Polyamine levels varied considerably between the mothers and tended to decrease over time. Putrescine was the minor polyamine, whereas spermidine and spermine contents were very similar. The concentrations of the three polyamines were significantly higher in hindmilk than foremilk (p < 0.001). Spermidine and spermine levels decreased significantly through the lactation progress (p < 0.05). Finally, slightly higher levels of polyamines were observed in the milk of mothers providing partial, rather than full, breastfeeding, although the differences were not significant. The polyamine content in human milk was found to change during a single feed (foremilk versus hindmilk) and as lactation progressed, mainly in response to the specific circumstances of the newborn.

Asunto(s)

Lactancia Materna/métodos , Leche Humana/química , Poliaminas/análisis , Adolescente , Adulto , Factores de Edad , Peso al Nacer , Índice de Masa Corporal , Cromatografía Líquida de Alta Presión , Estudios de Cohortes , Parto Obstétrico/métodos , Femenino , Humanos , Lactante , Recién Nacido , México , Madres , Poliaminas/química , Putrescina/análisis , Espermidina/análisis , Espermina/análisis , Adulto Joven

RESUMEN

Lulo (Solanum quitoense Lam.) is a Colombian fruit that is mostly used in the preparation of homemade juice as well as natural remedy for hypertension. The aim of this study was to determine physicochemical and antioxidant properties (antioxidant capacity, total phenols, flavonoids and spermidine content, and polyphenolic compounds profile by liquid chromatography-mass spectrometry (LC-MS)) of the lulo fruit and its juice. Additionally, vacuum impregnation (VI) properties of the fruit and the effect of high homogenization pressure (50, 100, and 150 MPa) on the juice properties were studied. The results revealed a good availability and impregnation capacity of the pores in fruits with similar maturity index. The main differences observed between the juice and fruit derive from removing solids and bioactive components in the filtering operation. However, the effect of high-pressure homogenization (HPH) on particle size and bioactive compounds increases the antiradical capacity of the juice and the diversity in polyphenolics when increasing the homogenization pressure.

RESUMEN

Cloning techniques are used to improve agronomical traits and answer to the demand for fine chocolate. The objective of this study was to predict the concentrations of bioactive amines, phenolic compounds, and the antioxidant potential of dark monoclonal chocolate from nine fine cocoa varieties by FTIR analysis and conventional techniques. Total phenolic compounds, bioactive amines and antioxidant activity varied significantly among chocolates. The antioxidant activity was also affected by the analytical method (DPPH vs. Rancimat). Chemometric models based on FTIR data provided satisfactory predictions of the concentrations of the amines: spermidine (R2 = 0.92; RMSEP = 0.39; RMSEC = 0.21), tryptamine (R2 = 0.92; RMSEP = 0.41; RMSEC = 0.20), cadaverine (R2 = 0.82; RMSEP = 1.58; RMSEC = 0.75) and tyramine (R2 = 0.87; RMSEP = 1.87; RMSEC = 0.68); as well as phenolic compounds and antioxidant activity by Rancimat® (R2 = 0.98; RMSEP = 0.32; RMSEC = 0.21) and DPPH (R2 = 0.97; RMSEP = 4.05; RMSEC = 1.66). The wavenumbers of amines vibrations are among those that most affected antioxidant prediction models, confirming the contribution of amines to the antioxidant activity of chocolates.

RESUMEN

Passiflora setacea is a wild species of passion fruit with interesting functional properties. Fruit seasonality demands conservation methods to enable its consumption throughout the year. We evaluated High Temperature Short Time (HTST) and Low Temperature Long Time (LTLT) binomials on physical, chemical, antioxidant and sensory characteristics of Passiflora setacea pulps. In natura (IN) and pasteurized pulps were analysed for DPPH, FRAP, ORAC, total phenolic content (TPC), vitamin C, bioactive amines, flavonoids, color, remaining enzymatic activity (REA), microbiological analyzes, sensory evaluation and physical stability. All binomials reached microbiological standards. Binomials 82 °C/20 s and 82 °C/40 s were selected for providing higher total antioxidant activity (TAA), TPC and lower REA. The highest levels of antioxidant activity, flavonoids, vitamin C were kept by 82 °C/20 s, without difference from IN pulp. LTLT binomial showed higher retention of bioactive amines, but also higher REA. Sensory acceptance was not affected by the binomials but pasteurized-cooked flavor was more checked for 82 °C 40 s than IN pulp.

RESUMEN

In nitrogen-fixing rhizobia, emerging evidence shows significant roles for polyamines in growth and abiotic stress resistance. In this work we show that a polyamine-deficient ornithine decarboxylase null mutant (odc2) derived from Sinorhizobium meliloti Rm8530 had significant phenotypic differences from the wild-type, including greatly reduced production of exopolysaccharides (EPS; ostensibly both succinoglycan and galactoglucan), increased sensitivity to oxidative stress and decreased swimming motility. The introduction of the odc2 gene borne on a plasmid into the odc2 mutant restored wild-type phenotypes for EPS production, growth under oxidative stress and swimming. The production of calcofluor-binding EPS (succinoglycan) by the odc2 mutant was also completely or mostly restored in the presence of exogenous spermidine (Spd), norspermidine (NSpd) or spermine (Spm). The odc2 mutant formed about 25 % more biofilm than the wild-type, and its ability to form biofilm was significantly inhibited by exogenous Spd, NSpd or Spm. The odc2 mutant formed a less efficient symbiosis with alfalfa, resulting in plants with significantly less biomass and height, more nodules but less nodule biomass, and 25 % less nitrogen-fixing activity. Exogenously supplied Put was not able to revert these phenotypes and caused a similar increase in plant height and dry weight in uninoculated plants and in those inoculated with the wild-type or odc2 mutant. We discuss ways in which polyamines might affect the phenotypes of the odc2 mutant.

Asunto(s)

Medicago sativa/microbiología , Ornitina Descarboxilasa/genética , Poliaminas/metabolismo , Nódulos de las Raíces de las Plantas , Sinorhizobium meliloti/genética , Proteínas Bacterianas/genética , Medicago sativa/crecimiento & desarrollo , Medicago sativa/metabolismo , Mutación , Nitrógeno/metabolismo , Fenotipo , Polisacáridos Bacterianos/metabolismo , Nódulos de las Raíces de las Plantas/metabolismo , Nódulos de las Raíces de las Plantas/microbiología , Sinorhizobium meliloti/metabolismo

RESUMEN

RATIONALE: Individuals with opioid use disorders often relapse into drug-seeking behavior after recalling memories linked to the drug use experience. Improving extinction efficacy has been used as a strategy to treat substance use disorders and suppress relapse. Although N-methyl-D-aspartate receptor (NMDAr) agonists facilitate acquisition, consolidation, and extinction, no study has addressed whether spermidine (SPD), a natural polyamine ligand of the NMDA receptor, facilitates the extinction and reinstatement of morphine-induced conditioned place preference (CPP). OBJECTIVES AND METHODS: The aim of the present study was to investigate the effect of SPD, an NMDAr agonist, on the extinction and reinstatement of morphine-induced CPP in mice. Adult male albino Swiss mice received saline (0.9% NaCl) or morphine (5 mg/kg) intraperitoneally (i.p.) and were respectively confined to a black or a white compartment for 30 min for four consecutive days for CPP induction. SPD (10-30 mg/kg, i.p.) or ifenprodil (NMDAr antagonist, 0.1-1 mg/kg, i.p.) were injected 15 min before extinction training. RESULTS: SPD and ifenprodil facilitated the extinction of morphine-induced CPP. SPD treatment during the extinction period impaired reinstatement induced by a priming dose of morphine (1.25 mg/kg). Ifenprodil (0.1 mg/kg) prevented the facilitatory effect of spermidine on the extinction of morphine-induced CPP but did not prevent reinstatement induced by morphine. CONCLUSIONS: These results suggest that SPD facilitated the extinction of morphine-induced CPP by modulating the polyamine binding site of the NMDA receptor. Our findings reveal important effects of SPD and ifenprodil on the re-exposure-induced decrease in morphine-induced CPP, which may be promising for developing novel pharmacological strategies to treat opioid use disorder.

Asunto(s)

Condicionamiento Clásico/efectos de los fármacos , Comportamiento de Búsqueda de Drogas/efectos de los fármacos , Extinción Psicológica/efectos de los fármacos , Morfina/efectos adversos , Receptores de N-Metil-D-Aspartato/agonistas , Espermidina/uso terapéutico , Animales , Condicionamiento Clásico/fisiología , Comportamiento de Búsqueda de Drogas/fisiología , Extinción Psicológica/fisiología , Masculino , Ratones , Morfina/farmacología , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , N-Metilaspartato/farmacología , Trastornos Relacionados con Opioides/tratamiento farmacológico , Trastornos Relacionados con Opioides/psicología , Receptores de N-Metil-D-Aspartato/metabolismo , Espermidina/farmacología

RESUMEN

Chagas´ disease continues to be a challenging and neglected public health problem in many American countries. The etiologic agent, Trypanosoma cruzi, develops intracellularly in the mammalian host, which hinders treatment efficacy. Progress in the knowledge of parasite biology and host-pathogen interaction has not been paralleled by the development of novel, safe and effective therapeutic options. It is then urgent to seek for novel therapeutic candidates and to implement drug discovery strategies that may accelerate the discovery process. The most appealing targets for pharmacological intervention are those essential for the pathogen and, whenever possible, absent or significantly different from the host homolog. The thiol-polyamine metabolism of T. cruzi offers interesting candidates for a rational design of selective drugs. In this respect, here we critically review the state of the art of the thiolpolyamine metabolism of T. cruzi and the pharmacological potential of its components. On the other hand, drug repurposing emerged as a valid strategy to identify new biological activities for drugs in clinical use, while significantly shortening the long time and high cost associated with de novo drug discovery approaches. Thus, we also discuss the different drug repurposing strategies available with a special emphasis in their applications to the identification of drug candidates targeting essential components of the thiol-polyamine metabolism of T. cruzi.

Asunto(s)

Reposicionamiento de Medicamentos/métodos , Poliaminas/metabolismo , Compuestos de Sulfhidrilo/metabolismo , Tripanocidas/farmacología , Trypanosoma cruzi/efectos de los fármacos , Animales , Antidepresivos/farmacología , Antipsicóticos/farmacología , Glutatión/análogos & derivados , Glutatión/biosíntesis , Humanos , Espermidina/análogos & derivados , Espermidina/biosíntesis , Trypanosoma cruzi/metabolismo

RESUMEN

Non-small cell lung cancer (NSCLC) is the most lethal and prevalent type of lung cancer. In almost all types of cancer, the levels of polyamines (putrescine, spermidine, and spermine) are increased, playing a pivotal role in tumor proliferation. Indomethacin, a non-steroidal anti-inflammatory drug, increases the abundance of an enzyme termed spermidine/spermine-N1-acetyltransferase (SSAT) encoded by the SAT1 gene. This enzyme is a key player in the export of polyamines from the cell. The aim of this study was to compare the effect of indomethacin on two NSCLC cell lines, and their combinatory potential with polyamine-inhibitor drugs in NSCLC cell lines. A549 and H1299 NSCLC cells were exposed to indomethacin and evaluations included SAT1 expression, SSAT levels, and the metabolic status of cells. Moreover, the difference in polyamine synthesis enzymes among these cell lines as well as the synergistic effect of indomethacin and chemical inhibitors of the polyamine pathway enzymes on cell viability were investigated. Indomethacin increased the expression of SAT1 and levels of SSAT in both cell lines. In A549 cells, it significantly reduced the levels of putrescine and spermidine. However, in H1299 cells, the impact of treatment on the polyamine pathway was insignificant. Also, the metabolic features upstream of the polyamine pathway (i.e., ornithine and methionine) were increased. In A549 cells, the increase of ornithine correlated with the increase of several metabolites involved in the urea cycle. Evaluation of the levels of the polyamine synthesis enzymes showed that ornithine decarboxylase is increased in A549 cells, whereas S-adenosylmethionine-decarboxylase and polyamine oxidase are increased in H1299 cells. This observation correlated with relative resistance to polyamine synthesis inhibitors eflornithine and SAM486 (inhibitors of ornithine decarboxylase and S-adenosyl-L-methionine decarboxylase, respectively), and MDL72527 (inhibitor of polyamine oxidase and spermine oxidase). Finally, indomethacin demonstrated a synergistic effect with MDL72527 in A549 cells and SAM486 in H1299 cells. Collectively, these results indicate that indomethacin alters polyamine metabolism in NSCLC cells and enhances the effect of polyamine synthesis inhibitors, such as MDL72527 or SAM486. However, this effect varies depending on the basal metabolic fingerprint of each type of cancer cell.

RESUMEN

Polyamines (PAs) are ubiquitous polycations derived from basic l-amino acids whose physiological roles are still being defined. Their biosynthesis and functions in nitrogen-fixing rhizobia such as Sinorhizobium meliloti have not been extensively investigated. Thin layer chromatographic and mass spectrometric analyses showed that S. meliloti Rm8530 produces the PAs, putrescine (Put), spermidine (Spd) and homospermidine (HSpd), in their free forms and norspermidine (NSpd) in a form bound to macromolecules. The S. meliloti genome encodes two putative ornithine decarboxylases (ODC) for Put synthesis. Activity assays with the purified enzymes showed that ODC2 (SMc02983) decarboxylates both ornithine and lysine. ODC1 (SMa0680) decarboxylates only ornithine. An odc1 mutant was similar to the wild-type in ODC activity, PA production and growth. In comparison to the wild-type, an odc2 mutant had 45 % as much ODC activity and its growth rates were reduced by 42, 14 and 44 % under non-stress, salt stress or acid stress conditions, respectively. The odc2 mutant produced only trace levels of Put, Spd and HSpd. Wild-type phenotypes were restored when the mutant was grown in cultures supplemented with 1 mM Put or Spd or when the odc2 gene was introduced in trans. odc2 gene expression was increased under acid stress and reduced under salt stress and with exogenous Put or Spd. An odc1 odc2 double mutant had phenotypes similar to the odc2 mutant. These results indicate that ODC2 is the major enzyme for Put synthesis in S. meliloti and that PAs are required for normal growth in vitro.

Asunto(s)

Ornitina Descarboxilasa/metabolismo , Poliaminas/metabolismo , Sinorhizobium meliloti/crecimiento & desarrollo , Sinorhizobium meliloti/metabolismo , Aminoácidos/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Medios de Cultivo , Regulación Bacteriana de la Expresión Génica , Prueba de Complementación Genética , Mutación , Ornitina Descarboxilasa/genética , Poliaminas/análisis , Putrescina/metabolismo , Sinorhizobium meliloti/enzimología , Espermidina/análogos & derivados , Espermidina/metabolismo , Transcripción Genética

RESUMEN

Among the main problems of some temperate fruit species, such as the apple tree (Malus domestica), are the poor set of fruits and low production. Polyamines and Self-Incompatibility Control Substances (SICS), involving mineral nutrients such as manganese and boron, are the major chemical compounds used to reduce these problems. The aim of this study was to use popular polyamines putrescine (Put) at 0.1, 0.25 mM, spermine (Spm) and spermidine (Spd) both at 0.05, 0.25 mM and SICS at 1, 2 mg L-1, alone or with cotton coverage bags and control to show the effects of these chemical compounds on yield indices and qualitative traits of apple (Malus domestica) cv ‘Red Delicious’. Results showed that Spd (0.25 mM) and SICS (1 mg L-1) had higher effect on yield per weight and per fruit number, final fruit set and ISI, but Spd (0.25 mM) decreased final drop. Put (0.1 mM + ccb), Spd (0.25 mM) and SICS (2 mg L-1 + ccb) were the most suitable treatments in order to increase the qualitative characteristics.

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Entre os principais problemas de algumas espécies frutíferas de clima temperado, como a macieira (Malus domestica), está o fraco conjunto de frutos e a baixa produção. As Poliaminas e Substancias de Controle de Auto-Incompatibilidade (self-incompatibility control substances-SICS) (envolvendo nutrientes minerais tais como manganês e boro) são os principais compostos químicos usados para reduzir esses problemas. O objetivo deste trabalho foi empregar a poliamina putrescina (Put) em 0,1, 0,25 mM, espermina (Spm) e espermidina (Spd) ambas em 0,05, 0,25 mM e SICS em 1,2 mg L-1, sozinhos ou com sacos cobertos de algodão, para demonstrar o efeito destes elementos químicos nos índices de produção e características qualitativas da maça (Malus domestica) ‘Red Delicious’. Resultados mostraram que a espermidina (0.25 mM) e SICS (1 mg L-1) tiveram um efeito maior na produção por peso e pelo número de frutos, o conjunto final de frutos e ISI, mas a espermidina (0.25 mM) caiu na produção final. Put (0.1 mM + sca), Spd (0.25 mM) e SICS (2 mg L-1 + sca) foram os tratamentos mais adequados para aumentar as características qualitativas.

RESUMEN

SIGNIFICANCE: Major pathogenic enterobacteria and protozoan parasites from the phylum Euglenozoa, such as trypanosomatids, are endowed with glutathione (GSH)-spermidine (Sp) derivatives that play important roles in signaling and metal and thiol-redox homeostasis. For some Euglenozoa lineages, the GSH-Sp conjugates represent the main redox cosubstrates around which entire new redox systems have evolved. Several proteins underwent molecular adaptations to synthesize and utilize the new polyamine-based thiols. Recent Advances: The genomes of closely related organisms have recently been sequenced, which allows mining and analysis of gene sequences that belong to these peculiar redox systems. Similarly, the three-dimensional structures of several of these proteins have been solved, which allows for comparison with their counterparts in classical redox systems that rely on GSH/glutaredoxin and thioredoxin. CRITICAL ISSUES: The evolutionary and structural aspects related to the emergence and use of GSH-Sp conjugates in Euglenozoa are reviewed focusing on unique structural specializations that proteins developed to use N1,N8-bisglutathionylspermidine (trypanothione) as redox cosubstrate. An updated overview on the biochemical and biological significance of the major enzymatic activities is also provided. FUTURE DIRECTIONS: A thiol-redox system strictly dependent on trypanothione is a feature unique to trypanosomatids. The physicochemical properties of the polyamine-GSH conjugates were a major driving force for structural adaptation of proteins that use these thiols as ligand and redox cofactor. In fact, the structural differences of indispensable components of this system can be exploited toward selective drug development. Future research should clarify whether additional cellular processes are regulated by the trypanothione system. Antioxid. Redox Signal. 28, 463-486.

Asunto(s)

Glutarredoxinas/genética , Compuestos de Sulfhidrilo/química , Tiorredoxinas/genética , Trypanosomatina/metabolismo , Evolución Molecular , Glutarredoxinas/química , Glutarredoxinas/metabolismo , Oxidación-Reducción , Poliaminas/química , Poliaminas/metabolismo , Espermidina/química , Espermidina/metabolismo , Compuestos de Sulfhidrilo/metabolismo , Tiorredoxinas/química , Tiorredoxinas/metabolismo , Trypanosomatina/química , Trypanosomatina/genética

RESUMEN

Among the main problems of some temperate fruit species, such as the apple tree (Malus domestica), are the poor set of fruits and low production. Polyamines and Self-Incompatibility Control Substances (SICS), involving mineral nutrients such as manganese and boron, are the major chemical compounds used to reduce these problems. The aim of this study was to use popular polyamines putrescine (Put) at 0.1, 0.25 mM, spermine (Spm) and spermidine (Spd) both at 0.05, 0.25 mM and SICS at 1, 2 mg L-1, alone or with cotton coverage bags and control to show the effects of these chemical compounds on yield indices and qualitative traits of apple (Malus domestica) cv ‘Red Delicious. Results showed that Spd (0.25 mM) and SICS (1 mg L-1) had higher effect on yield per weight and per fruit number, final fruit set and ISI, but Spd (0.25 mM) decreased final drop. Put (0.1 mM + ccb), Spd (0.25 mM) and SICS (2 mg L-1 + ccb) were the most suitable treatments in order to increase the qualitative characteristics.(AU)

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Entre os principais problemas de algumas espécies frutíferas de clima temperado, como a macieira (Malus domestica), está o fraco conjunto de frutos e a baixa produção. As Poliaminas e Substancias de Controle de Auto-Incompatibilidade (self-incompatibility control substances-SICS) (envolvendo nutrientes minerais tais como manganês e boro) são os principais compostos químicos usados para reduzir esses problemas. O objetivo deste trabalho foi empregar a poliamina putrescina (Put) em 0,1, 0,25 mM, espermina (Spm) e espermidina (Spd) ambas em 0,05, 0,25 mM e SICS em 1,2 mg L-1, sozinhos ou com sacos cobertos de algodão, para demonstrar o efeito destes elementos químicos nos índices de produção e características qualitativas da maça (Malus domestica) ‘Red Delicious. Resultados mostraram que a espermidina (0.25 mM) e SICS (1 mg L-1) tiveram um efeito maior na produção por peso e pelo número de frutos, o conjunto final de frutos e ISI, mas a espermidina (0.25 mM) caiu na produção final. Put (0.1 mM + sca), Spd (0.25 mM) e SICS (2 mg L-1 + sca) foram os tratamentos mais adequados para aumentar as características qualitativas.(AU)

RESUMEN

Polyamines are polycationic molecules that contain two or more amino groups (-NH3 +) and are present in all eukaryotic and prokaryotic cells. Polyamines are synthesized from arginine, ornithine, and proline, and from methionine as the methyl-group donor. In the traditional pathway for polyamine synthesis, arginase converts arginine into ornithine, which is decarboxylated by ornithine decarboxylase (ODC1) to generate putrescine. The latter is converted to spermidine and spermine. Recent studies have indicated the existence of 'non-classical pathways' for the generation of putrescine from arginine and proline in animal cells. Specifically, arginine decarboxylase (ADC) catalyzes the conversion of arginine into agmatine, which is hydrolyzed by agmatinase (AGMAT) to form putrescine. Additionally, proline is oxidized by proline oxidase to yield pyrroline-5-carboxylate, which undergoes transamination with glutamate to produce ornithine for decarboxylation by ODC1. Intracellular production of polyamines is controlled by antizymes binding to and inactivating ODC1. Polyamines exert effects that include stimulation of cell division and proliferation, gene expression for the survival of cells, DNA and protein synthesis, regulation of apoptosis, oxidative stress, angiogenesis, and cell-cell communication activity. Accordingly, polyamines are essential for early embryonic development and successful pregnancy outcome in mammals. In this paper the main concepts on the history, structure and molecular pathways of polyamines as well as their physiological role on angiogenesis, and reproductive physiology are reviewed.

Asunto(s)

Blastocisto/fisiología , Neovascularización Fisiológica/fisiología , Estrés Oxidativo/fisiología , Poliaminas/química , Poliaminas/metabolismo , Animales , Femenino , Humanos , Mamíferos , Embarazo , Ovinos