Decreased plasmatic spermidine and increased spermine in mild cognitive impairment and Alzheimer's disease patients

Abstract Background Current evidence suggests that upregulation of polyamines system plays a role both in cognitive deficit and synaptic loss observed in Alzheimer's disease (AD). Objective The aim of this study was to determine the plasmatic concentration of polyamines in mild cognitive impairment (MCI) and AD patients in comparison with healthy controls (HC). Methods Plasmatic polyamines were quantified using the AbsoluteIDQ® p180 and liquid chromatography coupled to tandem mass spectrometry (LC/MS-MS). Results The study group comprised 34 AD patients, 20 MCI and 25 HC. All individuals were followed for 4 years. During this period 8 amnestic MCI patients (40% of the MCI sample at baseline) converted to AD. Spermidine level was lower in both patient groups (AD; MCI) compared to HC (p = 0.007). Plasma levels of spermine were higher in the MCI group (p < 0.001), but decreased in the sub-sample of MCI patients who converted to AD (p = 0.043). No statistically significant differences were found in ornithine and putrescine levels (p = 0.056 and p = 0.126, respectively). Discussion Our results suggest dynamic changes in the expression of polyamines in the MCI-AD continuum.

Characterization of OAZ1 and its potential functions in goose follicular development

Background: Ornithine decarboxylase antizyme 1 (OAZ1) is an important regulator of polyamine synthesis and uptake. Our previous studies indicated that high OAZ1 expression in the ovaries of laying geese is responsible for poor egg production. In the present study, the molecular characterization of goose OAZ1 gene was analyzed, as well as the expression profile in various follicular tissues. Results: An 873-bp cDNA sequence of the OAZ1 gene (Accession No. KC845302) with a +1 frameshift site (+175T) was obtained. The sequence consisted of a 652-bp two overlapping open reading frames (a putative protein with 216 amino acids). The OAZ domain, OAZ signature and OAZ super family domain were prominent conserved regions among species. As the follicle size increased, OAZ1 abundance showed an increasing trend during follicular development, while it decreased during follicular regression. The level of OAZ1 mRNA expression was the lowest in the fifth largest preovulatory follicle, and was 0.65-fold compared to the small white follicle (P b 0.05). OAZ1 mRNA expression in the largest preovulatory and postovulatory follicle was 2.11- and 2.49-fold compared to the small white follicle, respectively (P b 0.05). Conclusions: The goose OAZ1 structure confirms that OAZ1 plays an important role in ornithine decarboxylase-mediated regulation of polyamine homeostasis. Our findings provide an evidence for a potential function of OAZ1 in follicular development, ovulation and regression.

Flavoproteínas que actúan como amino-oxidasas: Estructura, función e importancia clínica / Flavoproteins acting as amine oxidases: Structure, function and clinical significance / Flavoproteínas atuando como amina oxidases: Estrutura, função e significado clínico

Se destaca la actividad de las flavoenzimas como amino-oxidasas, que intervienen en el metabolismo de las aminas biogénicas como biorreguladores, especialmente en el crecimiento y la diferenciación celular. La clasificación de las amino-oxidasas incluye flavoenzimas y quinoenzimas. Se analizan las amino-oxidasas que son flavoproteínas, como las monoamino-oxidasas y las poliamino-oxidasas. Se discuten las isoformas, estructuras y función de ambas, sus sustratos e inhibidores, la expresión de MAO-A y MAO-B en tejidos humanos y sus implicancias clínicas. MAO plaquetaria es un biomarcador de desórdenes mentales y neurodegenerativos. Los inhibidores selectivos de MAO-A resultaron ser eficaces antidepresivos, mientras que algunos de MAO-B se utilizan en el tratamiento de enfermedades de Parkinson y de Alzheimer. La identificación de elevadas concentraciones de poliaminas en varias enfermedades, desde cáncer y psoriasis hasta infecciones parasitarias, hace que la manipulación de su metabolismo sea un blanco terapéutico o preventivo en ciertas enfermedades. Se discute además qué poliamino-oxidasas actúan en el metabolismo de las poliaminas en humanos, frente a las presentes en plantas, bacterias y protistas. Las poliaminas y las enzimas de su metabolismo desempeñan funciones relevantes en los procesos de envejecimiento y en algunas enfermedades, como cáncer, diabetes mellitus, accidentes cerebro-vasculares, insuficiencia renal y trastornos psiquiátricos.

The activity of flavoenzymes as amine oxidases involved in the metabolism of biogenic amines as bioregulators is highlighted, particularly for cell growth and differentiation. The classification of amine oxidases includes flavoenzymes and quinoenzymes. Amine oxidases that are flavoproteins, such as monoamine oxidases and polyamine oxidases, are analyzed herein. The isoforms, structures and functions of both enzyme families, their substrates and inhibitors, the expression of MAO-A and MAO-B in human tissues, and their clinical implications are discussed. Platelet MAO is a biomarker of mental and neurodegenerative disorders. Selective MAO-A inhibitors proved to be effective antidepressants, while some MAO-B inhibitors are used for treatment of Parkinson's and Alzheimer's diseases. The identification of high concentrations of polyamines in a variety of diseases, from psoriasis to cancer and parasitic infections, makes handling their metabolism a therapeutic or preventive target for the treatment of some diseases. Also polyamine oxidase activity on polyamine metabolism in humans, compared to those present in plants, bacteria and protists,is discussed. Polyamines and the enzymes involved in their metabolism play important roles in the aging processes, as well as in certain diseases such as cancer, diabetes mellitus, stroke, kidney failure, and defined psychiatric disorders.

Foi enfatizada a atividade de flavoenzimas como as amina oxidases envolvidas no metabolismo de aminas biogênicas como biorreguladores, especialmente no crescimento e diferenciação celular. A classificação das amina oxidases inclui flavoenzimas e quinoenzimas. Amina oxidases que são flavoproteínas, tais como monoamina oxidases e poliamina oxidases, são analisadas. Isoformas, estrutura e função das duas oxidases são discutidas, os seus substratos e inibidores, a expressão de MAO-A e MAO-B em tecidos humanos e suas implicações clínicas. MAO plaquetária é um biomarcador de desordens mentais e neurodegenerativas. Os inibidores selectivos da MAO-A resultaram ser eficazes antidepressivos, embora alguns dos MAO-B sejam utilizados no tratamento da doença de Parkinson e de Alzheimer. A identificação de elevadas concentrações de poliaminas em várias doenças, desde câncer e psoríase a infecções parasitárias, faz com que a manipulação do seu metabolismo seja um alvo terapêutico ou preventivo em certas doenças. Também se discute que a poliamina oxidase atua sobre o metabolismo das poliaminas no ser humano, em comparação com aquelas presentes em plantas, bactérias e protistas. As poliaminas e enzimas do seu metabolismo desempenham papéis relevantes nos processos de envelhecimento e em algumas doenças, tais como câncer, diabetes miellitus, acidente vascular cerebral, insuficiência renal e perturbações psiquiátricas.

Sodium butyrate does not decrease the evolution of precancerous lesions in rats / Butirato de sódio não diminui a evolução de lesões pré-neoplásicas em ratos

PURPOSE: To evaluate the preventive effect of sodium butyrate in the appearance of aberrant crypt foci (ACF) in rats after induction with the carcinogen 1,2-dimethylhydrazine (DMH). METHODS: Forty Wistar rats were separated into four groups (n=10) distributed as follows: control 1, control 2, butyrate 1 and butyrate 2. The groups control 1 and butyrate 1 remained under experimentation for 4 weeks, while the groups control 2 and butyrate 2 remained for 8 weeks. In the first four weeks, the animals of the control groups received water ad libitum and the animals of the butyrate groups received a sodium butyrate solution (3.4 percent) ad libitum. Injections of the drug 1,2-dimethylhydrazine were applied during the two first weeks of the experiment in all the animals, concurrently with the application of sodium butyrate. The large intestine of the animals was removed, for the analysis of the ACF and of the content of polyamines. The animal feces were collected for the analysis of the SCFA profile. RESULTS: The spermidine presented a higher concentration in the group butyrate 2 in comparison to the group control 2. There was a significant difference in the concentration value (µmol/mL) of acetate in comparison to the groups control 2 and butyrate 2. CONCLUSION: The use of sodium butyrate together with the induction of colorectal cancer was not effective in the prevention of the disease progression.

OBJETIVO: Avaliar o efeito preventivo do butirato de sódio no surgimento de focos de cripta aberrante (FCA) em ratos após a indução com o carcinógeno 1,2-dimetilhidrazina. MÉTODOS: Quarenta ratos foram divididos em quatro grupos, com dez animais em cada. Os grupos controle 1 e butirato 1 ficaram em experimentação por 4 semanas e os grupos controle 2 e butirato 2 por oito semanas. Nas primeiras quatro semanas, os animais dos grupos controle receberam água ad libitum e os animais dos grupos butirato receberam solução de butirato de sódio (3,4 por cento) ad libitum. Em todos os animais foram aplicadas quatro injeções subcutâneas da droga 1,2-dimetilhidrazina nas duas primeiras semanas, concomitante a administração do butirato de sódio. Foi retirado o intestino grosso dos animais, para análise dos FCA e do teor de poliaminas. As fezes dos animais foram recolhidas para análise do perfil de AGCC. RESULTADOS: A espermidina apresentou maior concentração no grupo butirato 2 em relação ao grupo controle 2. Foi encontrada diferença significativa no valor da concentração de acetato quando comparado os grupos controle 2 e butirato 2. CONCLUSÃO: A utilização do butirato de sódio concomitante à indução do câncer colorretal não se mostrou efetiva na prevenção da progressão da doença.

Trypanosoma cruzi: Transporte de metabolitos esenciales obtenidos del hospedador / Trypanosoma cruzi: Transport of essential metabolites acquired from the host

El Trypanosoma cruzi es el agente causal de la enfermedad de Chagas, endémica en Argentina y en toda América Latina. Presenta numerosas características metabólicas diferenciales respecto a sus hospedadores insectos y mamíferos. Algunas de estas diferencias fueron consecuencia de millones de años de adaptación al parasitismo en los cuales estos organismos protozoarios reemplazaron, a lo largo de su evolución, muchas rutas metabólicas de biosíntesis por sistemas de transporte de metabolitos desde el hospedador. En esta revisión se describen los avances en el conocimiento de los sistemas de transporte tanto bioquímicos como también de las moléculas involucradas en dichos procesos. Se aborda con especial énfasis los transportadores de aminoácidos y poliaminas de T. cruzi de la familia AAAP (Amino Acid/Auxin Permeases) ya que parece ser exclusiva de los tripanosomátidos. Teniendo en cuenta que estas moléculas se encuentran completamente ausentes en mamíferos podrían ser consideradas como potenciales blancos contra el Trypanosoma cruzi.

Trypanosoma cruzi is the etiological agent of Chagas disease, a disease endemic not only in Argentina but also in all of Latinamerica. T. cruzi presents several metabolic characteristics which are completely absent in its insect vectors and in mammalian hosts. Some of these differences were acquired after millions of years of adaptation to parasitism, during which this protozoan replaced many biosynthetic routes for transport systems. In the present review, we describe the advances in the knowledge of T. cruzi transport processes and the molecules involved. In particular, we focus on aminoacid and polyamine transporters from the AAAP family (Amino Acid/Auxin Permeases), because they seem to be exclusive transporters from trypanosomatids. Taking into account that these permeases are completely absent in mammals, they could be considered as a potential target against Trypanosoma cruzi.

Antisense expression of a gene encoding a calcium-binding protein in transgenic tobacco leads to altered morphology and enhanced chlorophyll.

Entamoeba histolytica contains a novel calcium-binding protein like calmodulin,which was discovered earlier,and we have reported the presence of its homologue(s)and a dependent protein kinase in plants.To understand the functions of these in plants,a cDNA encoding a calcium-binding protein isolated from Entamoeba histolytica (EhCaBP)was cloned into vector pBI121 in antisense orientation and transgenic tobacco plants were raised.These plants showed variation in several phenotypic characters,of which two distinct features,more greenness and leaf thickness,were inherited in subsequent generations.The increase in the level of total chlorophyll in different plants ranged from 60% to 70%.There was no major change in chloroplast structure and in the protein level of D1,D2,LHCP and RuBP carboxylase.These morphological changes were not seen in antisense calmodulin transgenic tobacco plants,nor was the calmodulin level altered in EhCaBP antisense plants.

Influence of cytokinins, auxins and polyamines on in vitro mass multiplication of cotton (Gossypium hirsutum L. cv. SVPR2).

In the present investigation, the influence of different forms of cytokinins, auxins and polyamines were tested for mass multiplication and regeneration of cotton. Initially, for the identification of effective concentration for multiple shoot induction, various concentrations of BAP, Kin and 2iP along with IAA and NAA were tested. Among tested concentrations, media fortified with MS salts; B5 vitamins; 30 g/l, glucose; 2.0 mg/l, 2iP; 2.0 mg/l, IAA and 0.7 % agar showed best response for multiplication of shoot tip explants (20 shoots per shoot tip explants). In nodal explants, maximum of 18.6 shoots were obtained in the media fortified with MS salts, B5 vitamins, 30 g/l, glucose, 2.0 mg/l, 2iP, 1.0 mg/l, NAA and 0.7 % agar. Effect of different concentrations of polyamines like spermidine and putrescine were also tested along with the above said multiplication media. Among the various treatments, 20 mg/l of putrescine showed best response and the multiple of shoots were increased to 26.5 shoots per shoot tip explants and 24.5 shoots per nodal explants. Elongation of shoots was achieved on multiple shoot induction medium. Significant number of roots were initiated in the medium supplemented with MS salts, vitamin B5 and IBA (2.0 mg/l). The frequency of root induction was increased by addition of, PVP (10 mg/l) along with root induction medium and after 2 weeks, the roots reached the maximum length of 22 cm. Further, these plantlets were hardened by using sand, soil and vermiculate in 1:1:1 ratio. The hardened plants were transferred to the environmental growth chamber for proper acclimatization. The hardened plants were then transferred to field for boll yielding and they exhibited 100% survival.

Intracellular polyamine pools, oligopeptide-binding protein A expression, and resistance to aminoglycosides in Escherichia coli

The role of intracellular free polyamine (putrescine and spermidine) pools in multiple resistance to aminoglycoside antibiotics was investigated among in vitro selected kanamycin-resistant Escherichia coli J53 mutants expressing diminished oligopeptide-binding protein (OppA) levels and/or defective ornithine decarboxylase (ODC) activity. The results suggest that diminished OppA content, but not defective ODC activity expression, increased the relative concentration of free spermidine as compared to the wild type strain. Moreover, by adding exogenous polyamines or polyamine synthesis inhibitors to cultures with different mutant strains, a direct relationship between the intracellular OppA levels and resistance to kanamycin was revealed. Collectively these results further suggest a complex relation among OppA expression, aminoglycoside resistance and polyamine metabolism.

Melatonin reduces ultraviolet-B induced cell damages and polyamine levels in human skin fibroblasts in culture

UV radiation is known to cause photoaging of the skin and is considered one of the leading cause of developing skin carcinogenesis. Melatonin which has a highly lipophilic molecular structure facilitating penetration of cell membranes and serving as an extra- and intracellular free radical scavenger has been demonstrated to protect photodamage of skin affected by UV exposure. In this study, we have examined the role of melatonin in response to UVB induced photodamaging process, using human skin fibroblasts in vitro. Cell survival curves after UVB irradiation showed dose-dependent decrease. Only 60% of fibroblasts were survived at 140 mJ/cm2 UVB irradiation. By pre-cultivation of cells with melatonin (100 nM), a significant number of cells remained unaffected. After UVB irradiation with 70 mJ/cm2, the level of putrescine was 1.7+/-0.3 fold increased compared to melatonin pre-treated group. In Northern analyses, the transcriptional level of ornithine decarboxylase (ODC) gene expression was increased by UVB irradiation and prohibited by melatonin. These results indicated that melatonin was effectively able to neutralize membrane peroxidation when present in relevant concentration during UVB irradiation and diminishes the UVB-induced increase of polyamine synthesis and ODC gene expression. Collectively, ODC response to UVB induced changes are possibly involves a melatonin or antioxidant sensitive regulatory pathway in normal human skin fibroblast.

Changes in free polyamines and related enzymes during stipule and pod wall development in Pisum sativum.

Level of free polyamines, their key metabolic enzymes, and other features related to ageing were examined during stipule and pod wall development in pea (Pisum sativum). Free polyamine titre (per unit fresh mass) in both the organs, the specific activities of arginine decarboxylase and ornithine decarboxylase in the pod wall, gradually decreased with maturation. In stipule, these enzymes attained peak activity at 15 days after pod emergence and declined thereafter. Ornithine decarboxylase activity was greater in pod wall than in stipule; while, arginine decarboxylase activity was higher in stipule. Activity of degradative enzyme diamine oxidase increased with the onset of senescence in both the organs. Chlorophyll and electrical conductance had a inverse relationship throughout the experimental period, whereas, the chlorophyll content was directly related with polyamine levels in both stipule and pod wall during aging. On the other hand, protein and RNA contents were positively correlated with free polyamines throughout the test period in stipule, but in the pod wall this was true only for the later stages of development.

Role of polyamines and ethylene as modulators of plant senescence.

Under optimal conditions of growth, senescence, a terminal phase of development, sets in after a certain physiological age. It is a dynamic and closely regulated developmental process which involves an array of changes at both physiological and biochemical levels including gene expression. A large number of biotic and abiotic factors accelerate the process. Convincing evidence suggests the involvement of polyamines (PAs) and ethylene in this process. Although the biosynthetic pathways of both PAs and ethylene are interrelated, S-adenosylmethionine (SAM) being a common precursor, their physiological functions are distinct and at times antagonistic, particularly during leaf and flower senescence and also during fruit ripening. This provides an effective means for regulation of their biosynthesis and also to understand the mechanism by which the balance between the two can be established for manipulating the senescence process. The present article deals with current advances in the knowledge of the interrelationship between ethylene and PAs during senescence which may open up new vistas of investigation for the future.

Possible role of polyamines in gyrate atrophy.

PURPOSE: Gyrate atrophy (GA) is marked by hyperornithinemia and lowered ornithine amino transferase (OAT). However there are patients of GA without hyperornithinemia and those with hyperornithinemia without GA. Some cases of GA have been reported to have low lysine. The purpose of the study was to determine if polyamines, the metabolites of ornithine, and lysine have any diagnostic role in GA. METHODS: Ornithine in plasma was estimated by two-dimensional paper chromatography, with elution of the coloured spot, and the absorbance measured using a spectrophotometer at 560 nm. OAT assay in lymphocytes was done spectrophotometrically using ornithine as substrate. Blood and urinary polyamines were extracted with n-butanol, benzoylated and analysed with HPLC; putrescine, spermine, spermidine, and cadaverine were assayed individually at 254 nm with the UV detector using ODS, G18 column with 63% methanol as solvent. RESULTS: Of the 7 patients investigated, 6 had features typical of GA. One was diagnosed to have atypical retinitis pigmentosa (case 3). The first five cases had elevated ornithine and diminished OAT, but cases 6 and 7 had near-normal ornithine and case 7 had near-normal OAT. However, all 7 patients had increased levels of total polyamines in urine compared to normals. Five had increased putrescine and three had increased spermine. All the 7 had decreased cadaverine in urine. Thus, though there were inconsistencies with ornithine and OAT, all the 7 patients had elevated polyamines from ornithine and decreased cadaverine. CONCLUSION: In addition to estimating ornithine and OAT in GA, it is suggested that urinary polyamines may be analysed as the latter appears to correlate better with the clinical condition and help in the diagnosis to a greater extent. Moreover, while ornithine is an innocuous amino acid, polyamines are known to damage DNA and proteins.

Neurotransmisores y esquizofrenia / Neurotransmitters and schizophrenia

En la patologia esquizofrénica, los modelos bioquímicos explicativos mas comunes son: El de la Dopamina y el de la Fenciclidina. El neurotransmisor Dopamina actúa sobre los diversos receptores específicos, y la Fenciclidina sobre los llamados receptores de aminoácidos excitatorios. Otra hipótesis, como la de la Glicina o la de las Poliaminas, también tienen que ver con la acción sobre receptores de aminoacidos excitatorios. Esta revisión estudia algunos eventos neuroquímicos y neuropatológicos relacionados con los modelos mencionados. Conclusion: La esquizofrenia no puede ser explicada por un modelo simple, y mas bien es el resultado de una compleja interacción de disfunciones en los sistemas de neurotransmisión.

Efecto de L-arginina y poliaminas sobre la movilidad espermática / Effect of L-arginine and polyamines on spermatic motility

En este trabajo se destaca la importancia bioquímica y fisiológica en el sistema reproductor masculino de L-arginina, ya que este aminoácido ha sido utilizado con éxito en el tratamiento de algunos problemas de fertilidad. Este aminoácido está relacionado con el metabolismo de las poliaminas putrecinas, espermidina y espermina, moléculas que tienen importancia fisiológica, ya que se ha sugerido su participación como iniciadoras del proceso de movilidad espermática. Se sugieren algunas aplicaciones prácticas de la L-arginina.

Control of zoopathogenic fungi in vitro by polyamine biosynthesis inhibitors.

Effect of inhibitors of polyamine (PA) biosynthesis, alpha-difluoromethylornithine (DFMO), methylglyoxal bis (guanylhydrazone)--MGBG and bis (cyclohexylammonium) sulphate (BCHA) on mycelial growth of three clinically important fungi-Trichophyton mentagrophytes, Microsporum gypseum and Aspergillus flavus was examined in vitro. All inhibitors at concentrations 1 to 50 mM produced greater inhibition of mycelial growth in all fungi tested in a dose-dependent manner. MGBG was the most effective inhibitor, and T. mentagrophytes was the most sensitive fungus to all inhibitors followed by M. gypseum and A. flavus. The results suggested that control of fungal diseases in animals and human beings with specific inhibitors of PA biosynthesis is possible.

Insecticidal activity of inhibitors of polyamine biosynthesis on Spodoptera litura F. larvae.

The inhibitors of polyamine (PA) biosynthesis such as alpha-difluoromethylornithine (DFMO), methylglyoxal bis (guanylhydrazone) (MGBG) and bis (cyclohexylammonium) sulphate (BCHA) have been used to protect crop plants from pathogenic fungi. In this communication the insecticidal activity of these inhibitors on tobacco caterpillar, S. litura has been reported. All the inhibitors exhibited insecticidal activity; MGBG being more effective than others. The results suggest, for the first time, a possible avenue for the control of insect pests by specific inhibition of insect PA biosynthesis.

Action of methylglyoxal bis (guanyl hydrazone) and related antiprotozoals on Acanthamoeba culbertsoni.

Methylglyoxal bis(guanyl hydrazone) (MGBG) and the related diamidine compounds berenil and pentamidine inhibited multiplication of A. culbertsoni. The growth inhibition by MGBG (2.5 mM) in the peptone medium was accompanied by the disappearance of spermidine and a marked reduction in the level of diaminopropane. MGBG and berenil completely inhibited growth in a chemically defined medium at 1 mM and 1-2 microM concentration, respectively. However, there was no decrease in the polyamine levels in the early stages of growth inhibition by these agents. Uptake of putrescine, spermidine and spermine by A. culbertsoni has been demonstrated but addition of exogenous polyamines did not reverse the growth inhibitory action of MGBG and berenil. Inhibition of S-adenosylmethionine decarboxylase and decrease in polyamine synthesis do not seem to be the primary targets for the antiamoebic action of MGBG and berenil.

Comparative evaluation of sequestration of polyamines by perfused rabbit and rat lungs.

Differences were observed in the sequestration of polyamines putrescine, spermidine and spermine by isolated, ventilated, perfused rat and rabbit lungs, former being able to accumulate more polyamines compared to the latter. Steady state equilibrium was reached earlier for spermine in rat. Isolated ventilated lungs were perfused with harmaline and ouabain, inhibitors known to inhibit the sodium pump at a maximum concentration of 1 mM for rabbit lungs and 0.4 and 0.2 mM for rat lungs, respectively. They did not affect the uptake of polyamines by rat lung but decreased the uptake of putrescine by rabbit lung. Decreased sodium (50 meq/L) in the perfusate increased the uptake of spermine and spermidine by rabbit lung but again showed no effect with rat lung. However, the uptake of polyamines by isolated ventilated rat and rabbit lungs perfused for 60 min with these compounds was linear over the entire range of high concentrations studied. These results suggest that the major uptake process of polyamines by intact lungs of both animal species is primarily by simple diffusion. HPLC analysis of the perfusate and lungs from both animal species post-perfusion indicated no detectable metabolites of the polyamines.