RESUMO

In plant tissue culture, differences in endogenous levels of species-specific plant growth regulators (PGRs) may explain differences in regenerative capacity. In the case of polyamines (PAs), their dynamics and distribution may vary between species, genotypes, tissues, and developmental pathways, such as sexual reproduction and apomixis. In this study, for the first time, we aimed to assess the impact of varying endogenous PAs levels in seeds from distinct reproductive modes in Miconia spp. (Melastomataceae), on their in vitro regenerative capacity. We quantified the free PAs endogenous content in seeds of Miconia australis (obligate apomictic), Miconia hyemalis (facultative apomictic), and Miconia sellowiana (sexual) and evaluated their in vitro regenerative potential in WPM culture medium supplemented with a combination of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The morphogenic responses were characterized by light microscopy and scanning electron microscopy and discussed regarding the endogenous PAs profiles found. Seeds of M. sellowiana presented approximately eight times more putrescine than M. australis, which was associated with a higher percentage of regenerated calluses (76.67%) than M. australis (5.56%). On the other hand, spermine levels were significantly higher in M. australis. Spermine is indicated as an inhibitor of auxin-carrying gene expression, which may have contributed to its lower regenerative capacity under the tested conditions. These findings provide important insights into in vitro morphogenesis mechanisms in Miconia and highlight the significance of endogenous PA levels in plant regeneration. These discoveries can potentially optimize future regeneration protocols in Miconia, a plant group still underexplored in this area.

Assuntos

RESUMO

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.

Assuntos

RESUMO

Necro-leachate, a liquid released during cadaveric decomposition, is considered the main culprit for impacts on cemetery environments. The biogenic amines cadaverine and putrescine make up part of the composition of necro-leachate and have a certain toxicity to different organisms. Springtails are among the most used bioindicators to assess the impacts of soil contaminants. As there are no data on the acute and chronic toxicity of springtails exposed to cadaverine and putrescine, the objective of this study was to evaluate the toxic potential of both amines, under the behavioral effect of avoidance and reproduction in the species Folsomia candida. Springtails were exposed to soils contaminated with different concentrations of cadaverine and putrescine, and different mixtures of these amines. To evaluate the avoidance and reproduction test, the individuals were exposed for periods of 48 h and 28 days, respectively. The results obtained in the avoidance test showed that springtails exhibited a preference for the treated soil in both isolated and mixed treatments. The chronic evaluation assays showed that the reproduction was affected, particularly in the treatments with combined amines, resulting in a reduction in the total number of juveniles. From the results, it is possible to infer that the methods applied in this research have provided information that will contribute to a better understanding of the toxicity of putrefactive biogenic amines, since there exist few ecotoxicological studies carried out with these amines, and especially with those from cemetery environments.

Assuntos

RESUMO

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.

RESUMO

Cemetery leachate generated by the process of cadaveric decomposition is a significant contaminant of several matrices in the cemetery environment (soil, groundwater, and surface water). The biogenic amines cadaverine and putrescine stand out among the cemetery leachate contaminants, since they are potentially carcinogenic compounds. This review article presents a discussion of possible environmental impacts caused by the increase in deaths resulting from COVID-19 as its central theme. The study also aims to demonstrate the importance of considering, in this context, some climatic factors that can alter both the time of bodily decomposition and the longevity of the virus in the environment. Additionally, some evidence for the transmission of the virus to health professionals and family members after the patient's death and environmental contamination after the burial of the bodies will also be presented. Several sources were consulted, such as scientific electronic databases (NCBI), publications by government agencies (e.g., ARPEN, Brazil) and internationally recognized health and environmental agencies (e.g., WHO, OurWorldInData.org), as well as information published on reliable websites available for free (e.g., CNN) and scientific journals related to the topic. The data from this study sounds the alarm on the fact that an increase in the number of deaths from the complications of COVID-19 has generated serious environmental problems, resulting from Cemetery leachate.

Assuntos

RESUMO

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.

RESUMO

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.

Assuntos

RESUMO

The aim of this study was to evaluate the effects of increasing doses of putrescine injected in ovo on hatchability, intestinal morphology and pre-starter performance of broilers. For this purpose, 720 eggs from broiler breeders were separated into a negative control (no injection) and injection treatments with increasing doses of putrescine (0.05; 0.1; 0.15 and 0.2%), totalling five treatments of 144 eggs each. Eggs were distributed in a completely randomized design inside the setter and the injection of solutions occurred at 17 days of incubation. After hatch, 330 birds were housed in mixed lots following the original treatments, totalling 5 treatments of 6 replicates with 11 birds each. Six birds per treatment were weighed and euthanized by cervical dislocation to collect the liver, intestine and breast 24 hr after injection, at hatch and 24 hr after hatch. At 2 days of age, intestines were collected from 4 animals per treatment to analyse histomorphology. The effects of putrescine levels were evaluated by polynomial regression models, ANOVA and Tukey test at 5% probability. The hatchability decreased linearly in response to increased doses of putrescine. The percentage of residual yolk was lower in animals that received putrescine compared to the control. After injection, the percentage of breast increased linearly, and the percentage of intestine had a quadratic response to increased doses of putrescine. However, 24 hr after hatch, the percentage of intestine linearly decreased, and the percentage of liver linearly increased in response to increased doses of putrescine. Villus height increased quadratically, crypt depth decreased linearly, and goblet cells increased linearly in response to the putrescine dose. FI and BWG were not affected in the pre-starter phase; however, FCR increased in response to increased levels of putrescine. Due to putrescine effects on embryos, it is recommended that the doses injected in ovo not exceed 0.1%.

Assuntos

RESUMO

ABSTRACT In this study two experiments were carried out, the effect of a diet supplemented with fructans of Agave tequilana Weber, was analyzed in Hy-line hens in regards to their egg quality and production as well as on the levels of the polyamines, putrescine, spermidine and spermine in their feces. In the first study 300 Hy-line W-36 hens per group, which were aged 36 weeks, were randomly separated as follows for each of the three treatments. One group of 100 hens was fed with a diet supplemented with 0.1% fructans. Another group of 100 hens was supplemented with 0.2% fructans and a group of 100 hens without any fructans was added as a control group. Feed consumption was lower in the supplemented groups compared to the control group (p 0.05). Egg yolk quality was measured using Haugh units. The quality of the shell was studied using an Egg Force reader (g pressure/mm2). Putrescine levels was measured in 10 animals in each group. In the second study, 1,155 laying hens, aged one day from the Hy-line W-35 genetic line were also randomly divided into three groups containing 385 hens in each one (A, B, C). Egg laying levels and weight was measured during 35 weeks. The egg laying percentage increased considerably in hens ingesting supplemented diets (p 0.05) and the egg weight was greater mainly in the supplementation with 0.1% fructans (p 0.05). Thus, it was shown that diet supplementation with fructans of agave improves egg quality and homeostasis and food consumption in the Hy-line hen.

RESUMO

During plant-microbe interactions, polyamines participate in the plant defense response. Previously, we reported that silencing of ADC genes in Arabidopsis thaliana causes a drastic reduction of polyamine levels as well as increments in reactive oxygen species content. In this study, we examined the response of the adc-silenced line to Botrytis cinerea and Pseudomonas syringae infection. The adc-silenced line was more susceptible to Botrytis cinerea, showing larger lesion length and a higher incidence of fungal infection. Pre-treatments with putrescine reestablished the response of the adc-silenced line to Botrytis cinerea, resulting in a similar phenotype to the parental plant. Expression levels of defense-related genes were analyzed during fungal infection showing that the salicylic acid-induced gene PR1 was up-regulated, while the jasmonic acid-related genes LOX3 and PDF1.2, as well as, the camalexin biosynthetic gene PAD3 were down-regulated in the adc-silenced line. Furthermore, methyl jasmonate pre-treatments reduced Botrytis cinerea infection in the adc-silenced line. On the other hand, the adc-silenced line showed an increased resistance to Pseudomonas syringae infection. SA-related genes such as PR1, ZAT1.2, WRKY54 and WRKY70 were highly expressed in the adc-silenced line upon bacterial interaction. Our data show that the adc-silenced line has altered the defense-response against Botrytis cinerea and Pseudomonas syringae, that is consistent with deregulation of SA- and JA-mediated response pathways.

Assuntos

RESUMO

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.

RESUMO

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.

Assuntos

RESUMO

Chagas disease is a neglected tropical disease endemic to Latin America, though migratory movements have recently spread it to other regions. Here, we have applied a cascade virtual screening campaign combining ligand- and structure-based methods. In order to find novel inhibitors of putrescine uptake in Trypanosoma cruzi, an ensemble of linear ligand-based classifiers obtained by has been applied as initial screening filter, followed by docking into a homology model of the putrescine permease TcPAT12. 1,000 individual linear classifiers were inferred from a balanced dataset. Subsequently, different schemes were tested to combine the individual classifiers: MIN operator, average ranking, average score, average voting, with MIN operator leading to the best performance. The homology model was based on the arginine/agmatine antiporter (AdiC) from Escherichia coli as template. It showed 64% coverage of the entire query sequence and it was selected based on the normalized Discrete Optimized Protein Energy parameter and the GA341 score. The modeled structure had 96% in the allowed area of Ramachandran's plot, and none of the residues located in non-allowed regions were involved in the active site of the transporter. Positivity Predictive Value surfaces were applied to optimize the score thresholds to be used in the ligand-based virtual screening step: for that purpose Positivity Predictive Value was charted as a function of putative yields of active in the range 0.001-0.010 and the Se/Sp ratio. With a focus on drug repositioning opportunities, DrugBank and Sweetlead databases were subjected to screening. Among 8 hits, cinnarizine, a drug frequently prescribed for motion sickness and balance disorder, was tested against T. cruzi epimastigotes and amastigotes, confirming its trypanocidal effects and its inhibitory effects on putrescine uptake. Furthermore, clofazimine, an antibiotic with already proven trypanocidal effects, also displayed inhibitory effects on putrescine uptake. Two other hits, meclizine and butoconazole, also displayed trypanocidal effects (in the case of meclizine, against both epimastigotes and amastigotes), without inhibiting putrescine uptake.

Assuntos

RESUMO

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.

Assuntos

RESUMO

A highly sensitive method was developed to measure putrescine by micellar electrokinetic chromatography with laser induced fluorescence detection with excellent linearity in the 1 nM to 3 µM range. The technique was tested on a drop of blood from Parkinson's disease patients obtained by finger prick. The results showed a statistically significant increase of putrescine in the erythrocytes compared to controls and a non-significant increase in plasma. This high level of putrescine does not constitute by itself proof that putrescine and polyamines are directly related to Parkinson's disease. However, the present results and several others addressed in the discussion suggest that these compounds might be causally involved in the pathophysiology of Parkinson's disease. In addition, the analytical method reported here may help to find new biomarkers for many diseases including Parkinson's disease.

Assuntos

RESUMO

Infection with Trichomonas vaginalis produces a malodorous seropurulent vaginal discharge due to several chemicals, including polyamines. The presence of 1,4-diamino-2-butanone (DAB) reduces the amount of intracellular putrescine by 90%, preventing the cotransport of exogenous spermine. DAB-treated parasites present morphological changes, which are restored by adding exogenous putrescine into the culture medium. However, the effect of polyamines over the trichomonad proteomic profile is unknown. In this study, we used a proteomic approach to analyze the polyamine-depletion and restoration effect by exogenous putrescine on T. vaginalis proteome. In the presence of inhibitor DAB, we obtained 369 spots in polyamine-depleted condition and observed 499 spots in the normal culture media. With DAB treatment, the intensity of 43 spots was increased but was found to be reduced in 39 spots, as compared to normal conditions. Interestingly, in DAB-treated parasites restored with a medium with added exogenous putrescine, 472 spots were found, of which 33 were upregulated and 63 were downregulated in protein intensity. Some of these downregulated proteins in DAB-treated parasites are involved in several cellular pathways such as glycolysis, glycolytic fermentation, arginine dihydrolase pathway, redox homeostasis, host cell binding mediated by carbohydrate, chaperone function, and cytoskeletal remodeling. Interestingly, the intensity of some of the proteins was restored by adding exogenous putrescine. In conclusion, the presence of DAB altered the proteomic profile of T. vaginalis, resulting in a decrease in the intensity of 130 proteins and an increase in the intensity of 43 proteins that was restored by the addition of putrescine.

Assuntos

RESUMO

We have performed a biophysical characterization, at single molecule level, of the interaction between the DNA molecule and the biogenic polyamine putrescine. By using force spectroscopy, we were able to monitor the complexes formation as putrescine is added to the sample, determining the mechanical properties of such complexes and the physicochemical (binding) parameters of the interaction for three different ionic strengths. In particular, it was shown that the behavior of the equilibrium binding constant as a function of the counterion concentration deviates from the prediction of the Record-Lohman model. The measured constants were (1.3 ± 0.2) × 105 M- 1 for [Na] = 150 mM, (2.1 ± 0.2) × 105 M- 1 for [Na] = 10 mM, and (2.2 ± 0.3) × 105 M- 1 for [Na] = 1 mM. The cooperativity degree of the binding reaction, on the other hand, increases with the ionic strength. From these analysis, the DNA-putrescine binding mechanisms are inferred, and a comparison with results reported for ordinary bivalent ions like magnesium is performed. Such study provides new insights on the general behavior of the DNA interactions with biogenic polyamines.

Assuntos

RESUMO

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.

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.

RESUMO

Plants respond to pathogen attack by modifying defense gene expression and inducing the production of myriad proteins and metabolites. Among these responses, polyamine (PA) levels suffer remarkable modifications. Evidences demonstrate that plants make use of the polyamine biosynthetic pathway and the oxidative catabolism of these compounds in order to mount adequate defenses against pathogens. In Arabidopsis thaliana, putrescine is synthesized exclusively through the arginine decarboxylase (ADC) pathway, this enzyme exists as two isoforms named ADC1 and ADC2. Even though both isoforms participate in the response to pathogen attack, the mechanisms modulating ADC activity are not completely understood. Therefore, studies to clarify their roles are necessary. In this chapter, we describe the methods that can be applied for the study of plant-pathogen interactions using Arabidopsis adc mutant plants.

Assuntos

Assuntos