Amounts Of Polyamines Research Articles

High-performance liquid chromatography determination of polyamine contents in poultry based on precolumn benzoyl chloride derivatization

Polyamines are widely present in animal cells and perform important physiological and biochemical functions. Moderate intake of polyamines contributes to good health. Poultry is one of the common sources of polyamines in daily life. However, different tissues from poultry contain different amounts of polyamine. In this study, an efficient and accurate method for the determination of polyamines was developed by high-performance liquid chromatography (HPLC) with precolumn benzoyl chloride derivatization. The linearity of the polyamines was good between 10–400 nmol/mL, with the limit of detection (LOD, S/N = 3) and limits of quantification (LOQ, S/N = 10) were 0.37 and 1.23 nmol/mL. The recoveries of the method ranged from 94.73 % to 106.58 % with high accuracy (RSD < 7 %) and no significant matrix effects or interferences. The RSD of intra-day, inter-day and inter-batch repeatability in the range of 0.07–1.57 %, 0.08–1.03 % and 1.46–4.57 %. The method was successfully used to detect polyamines in poultry tissue, with putrescine, spermidine and spermine levels ranging from 3.24 to 29.85 mg/kg, 0.46 to 164.11 mg/kg and 63.39 to 489.57 mg/kg, respectively. Polyamine levels were higher in the spleen, thymus and bursa of fabricius and lower in the heart and muscle. Furthermore, the method was employed to study the effect of exogenous spermine on polyamine deposition in different tissues of geese. In summary, the method is suitable for the detection of polyamine in animal tissues.

Variations in Proline Content, Polyamine Profiles, and Antioxidant Capacities among Different Provenances of European Beech (Fagus sylvatica L.).

International provenance trials are a hot topic in forestry, and in light of climate change, the search for more resilient beech provenances and their assisted migration is one of the challenges of climate-smart forestry. The main aim of the study was to determine intraspecific variability in European beech (Fagus sylvatica L.) among 11 beech provenances according to total antioxidant capacities estimated by various assays, such as DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid), FRAP (ferric reducing antioxidant power) assay, and radical scavenging capacity against nitric oxide (RSC-NO assays), as well as osmolyte content, primarily individual polyamines (putrescine, spermidine, and spermine), and free proline content. Polyamine amounts were quantified by using HPLC coupled with fluorescent detection after dansylation pretreatment. The highest values for radical scavenger capacity assays (ABTS, DPPH, and FRAP) were measured in the German provenances DE47 and DE49. Also, the highest NO inhibition capacity was found in the provenance DE49, while the highest content of proline (PRO), total phenolic content (TPC), and total flavonoid content (TFC) was recorded in DE47. The Austrian AT56 and German provenance DE49 were most abundant in total polyamines. This research underlines the importance of the application of common antioxidant assays as well as osmolyte quantification as a criterion for the selection of climate-ready beech provenances for sustainable forest management.

Wheat germ, a byproduct of the wheat milling industry, as a beneficial source of anti-aging polyamines: A quantitative comparison of various forms.

Polyamines have received a lot of attention since the 1990s because of their anti-aging, anti-chronic disease, and proliferative effects. Wheat germ was reported as one of the natural sources of high polyamine, especially spermidine. The current study used three types of wheat germ: group A was industrially separated germ from whole grain, group B was the commercially available germinated wheat germ, and group C was manually separated wheat germ from germinated grain. The polyamine content of putrescine, spermidine, and spermine has been determined using a simplified isocratic LC-MS/MS method. An optimized extraction procedure was performed on all seven samples for obtaining a polyamine-enriched extract. The three dominant carbomylated polyamines were identified by analyzing the extracted samples in order to determine their relative abundance. Wheat germ powders contain the highest amount of polyamines (220-337 μg/g) of which spermidine is one of the most important. Germinated wheat grains, on the other hand, contain the least amount of this polyamine. The commercially available separated wheat germs are suggested as a good nutrition source of these polyamines.

Evaluation of the effect of urea foliar application on polyamine content, grain protein content, and the agronomic traits of bread wheat cultivars under dryland and supplemental irrigation conditions

The purpose of the experiment was to investigate the relationship between the accumulation of polyamines during the filling period of wheat grain with yield and grain protein under dryland and supplemental irrigation (SI) conditions and also the effect of foliar spraying of urea on the amount of polyamines in grain a factorial experiment was conducted based on randomized complete block design with three replications during two cropping seasons. The examined treatments included water availability (dryland and SI), wheat cultivars (Homa, Sardari, Rijaw, Ouhadi, and Azar2), and foliar application of urea (control and 4% urea foliar application). The results demonstrated that SI (10%) reduced and urea foliar (12%) application increased the percent of grain protein. SI reduced the contents of three PAs by 24%: spermine, spermidine, and putrescine. But urea foliar application increased the three PAs in the first and second years by 31% and 27% in dryland conditions and 12% and 4% in SI conditions, respectively. These results indicated that there is less need for PAs in SI conditions, and they thus exhibit poorer accumulation in the grain; in dryland conditions, however, their content increases with greater access to foliar nitrogen. Older cultivars Sardari and Azar 2 had more polyamines. According to the observed positive correlation between 1000-grain weight and the three PAs under stress conditions, PAs are probably involved in the improvement of grain filling, and urea foliar application can affect drought tolerance by increasing PA accumulation. Moreover, cadaverine exhibited a different function from the other PAs.

Levilactobacillus brevis with High Production of Putrescine Isolated from Blue Cheese and Its Application.

Polyamine intake has been reported to help extend the lifespan of animals. Fermented foods contain high concentrations of polyamines, produced by fermenting bacteria. Therefore, the bacteria, isolated from fermented foods that produce large amounts of polyamines, are potentially used as a source of polyamines for humans. In this study, the strain Levilactobacillus brevis FB215, which has the ability to accumulate approximately 200 µM of putrescine in the culture supernatant, was isolated from fermented foods, specifically the Blue Stilton cheese. Furthermore, L. brevis FB215 synthesized putrescine from agmatine and ornithine, which are known polyamine precursors. When cultured in the extract of Sakekasu, a byproduct obtained during the brewing of Japanese rice wine containing high levels of both agmatine and ornithine, L. brevis FB215 grew to OD600 = 1.7 after 83 h of cultivation and accumulated high concentrations (~1 mM) of putrescine in the culture supernatant. The fermentation product also did not contain histamine or tyramine. The Sakekasu-derived ingredient fermented by the food-derived lactic acid bacteria developed in this study could contribute to increasing polyamine intake in humans.

Metal- and Organ-Specific Response to Heavy Metal-Induced Stress Mediated by Antioxidant Enzymes' Activities, Polyamines, and Plant Hormones Levels in Populus deltoides.

Besides anthropogenic factors, climate change causes altered precipitation patterns that indirectly affect the increase of heavy metals in soils due to hydrological effects and enhanced leaching (i.e., Cd and Ni), especially in the vicinity of mines and smelters. Phytoextraction is a well-known, powerful "green" technique for environmental clean-up that uses plants to extract, sequester, and/or detoxify heavy metals, and it makes significant contributions to the removal of persistent inorganic pollutants from soils. Poplar species, due to their growth features, high transpiration rate, large biomass, and feasible reproduction represent great candidates for phytoextraction technology. However, the consequences of concomitant oxidative stress upon plant metabolism and the mechanism of the poplar's tolerance to heavy metal-induced stress are still not completely understood. In this study, cuttings of poplar species (Populus deltoides W. Bartram ex Marshall) were separately exposed to two heavy metals (Cd2+ and Ni2+) that were triple the maximum allowed amount (MAA) (according to national legislation). The aim of the study was to estimate the effects of heavy metals on: (I) the accumulation of free and conjugated polyamines, (II) plant hormones (including abscisic acid-ABA and indole-3-acetic acid-IAA), and (III) the activities of different antioxidant enzymes at root and leaf levels. By using the selected ion monitoring (SIM) mode of gas chromatography with mass spectrometry (GC/MS) coupled with the isotopically labeled technique, amounts of ABA and IAA were quantified, while polyamine amounts were determined by using high-performance liquid chromatography (HPLC) with fluorometric detection after derivatization. The results showed that P. deltoides responded to elevated concentrations of heavy metals in soils by exhibiting metal- and organ-specific tolerance. Knowledge about tolerance mechanisms is of great importance for the development of phytoremediation technology and afforestation programs for polluted soils.

Regulatory effect of polyamines and indole on expression of stress adaptation genes in &lt;i&gt; Escherichia coli &lt;/i&gt;

Background. Indole and polyamines are involved in the regulation of physiological processes in bacteria associated with adaptation to stress, biofilm formation, antibiotic tolerance, and bacterial persistence. However, the molecular targets and mechanisms of action of these metabolites are still poorly understood. In this work, we studied the effect of polyamines and indole on the expression of such genes as: rpoS, relA, and spoT, encoding regulators of the general stress responses and starvation; hns and stpA, encoding global regulators of gene expression; rmf, yqjD, hpf, raiA, rsfS, sra, ettA, encoding ribosome hibernation factors.The aim. To study the regulatory effects of polyamines and indole on the expression of these genes, which are responsible for the adaptation of Escherichia coli to stress.Materials and methods. We used strains of E. coli in this study. The amount of polyamines was studied by thin layer chromatography. The indole concentration was determined by high performance liquid chromatography. Gene expression was studied using real-time RT-PCR.Results. The addition of polyamines putrescine, cadaverine and spermidine to the medium stimulated the expression of all the studied genes. The maximal stimulation was observed at the stationary phase mostly. Putrescine and spermidine had the most significant effect. At 24 h of cultivation, an equimolar conversion of exogenous tryptophan into indole was showed. At this time, the expression of two genes – rmf and raiA – increased.Conclusions. We have shown that polyamines upregulate the expression of all the studied genes at the transcriptional level. The stimulating effect is specific for the phase of the batch culture and the type of polyamine. Indole has a positive effect on the expression of the rmf and raiA genes.

Biological solution conditions and flanking sequence modulate LLPS of RNA G-quadruplex structures.

Guanine-rich regions of DNA or RNA can form structures with two or more consecutive G-quartets called G-quadruplexes (GQ). Recent studies reveal the potential for these structures to aggregate in vitro. Here, we report effects of in vivo concentrations of additives-amino acids, nucleotides, and crowding agents-on the structure and solution behavior of RNAs containing GQ-forming sequences. We found that cytosine nucleotides destabilize a model GQ structure at biological salt concentrations, while free amino acids and other nucleotides do not do so to a substantial degree. We also report that the tendency of folded GQs to form droplets or to aggregate depends on the nature of flanking sequence and the presence of additives. Notably, in the presence of biological amounts of polyamines, flanking regions on the 5'-end of the RNA drive more droplet-like phase separation, while flanking regions on the 3'-end, as well as both the 5'- and 3'-ends, induce more condensed, granular structures. Finally, we provide an example of a biological sequence in the presence of polyamines and show that crowders such as PEG and dextran can selectively cause its phase separation. These findings have implications for the participation of GQS in LLPS in vivo.

The Chemical Environment at Maturation Stage in Pinus spp. Somatic Embryogenesis: Implications in the Polyamine Profile of Somatic Embryos and Morphological Characteristics of the Developed Plantlets.

Changes in the chemical environment at the maturation stage in Pinus spp. somatic embryogenesis will be a determinant factor in the conversion of somatic embryos to plantlets. Furthermore, the study of biochemical and morphological aspects of the somatic embryos could enable the improvement of somatic embryogenesis in Pinus spp. In the present work, the influence of different amino acid combinations, carbohydrate sources, and concentrations at the maturation stage of Pinus radiata D. Don and Pinus halepensis Mill. was analyzed. In P. radiata, the maturation medium supplemented with 175 mM of sucrose and an increase in the amino acid mixture (1,100 mgL–1 of L-glutamine, 1,050 mgL–1 of L-asparagine, 350 mgL–1 of L-arginine, and 35 mgL–1 of L-proline) promoted bigger embryos, with a larger stem diameter and an increase in the number of roots in the germinated somatic embryos, improving the acclimatization success of this species. In P. halepensis, the maturation medium supplemented with 175 mM of maltose improved the germination of somatic embryos. The increase in the amount of amino acids in the maturation medium increased the levels of putrescine in the germinated somatic embryos of P. halepensis. We detected significant differences in the amounts of polyamines between somatic plantlets of P. radiata and P. halepensis; putrescine was less abundant in both species. For the first time, in P. radiata and P. halepensis somatic embryogenesis, we detected the presence of cadaverine, and its concentration changed according to the species.

Amine variations in faecal content in the first weeks of life of newborns in relation to breast-feeding or infant formulas.

Breast-feeding is the ideal nutrition for a newborn's integral necessities. It seems crucial therefore to know its composition in order to provide suitable infant formula when required. Of these, polyamines (with lactation and the microbiota being its intestinal source) are involved in the development of gut epithelium and immunity. Safety concerns limit human intervention studies. Therefore, we studied the amounts of polyamines supplied by breast milk (varying among mothers) or infant formula feeding, up to day 30 postpartum, in the faeces of newborns. Independent samples (68) of breast milk from fifty-nine healthy Caucasian woman (day 0, 1, 3, 5, 7, 15 and 30 postpartum) who had natural deliveries after week 38, same-day faeces of newborns when available (eighty-one from breast milk and fifty-five from infant formula fed) and six infant formulas were collected and the polyamine content was determined by HPLC. In breast milk, polyamines and isoamylamine (a primary amine), with inter-individual variations, increased over time (with a higher content of spermidine; no other amines were present). Overall, they were much higher than in infant formula. By the 2nd week after birth, polyamines, cadaverine and tyramine, but not isoamylamine, were higher in the faeces of those fed infant formula compared with those fed breast milk. Cadaverine and tyramine could be used to predict the feeding type used for newborns. The differences in the content may be related to distinct colonisation of amine-producing bacteria, which can be established by polyamines. Further studies are required to determine the clinical utility of these findings.

Putrescine enhances salt tolerance of cucumber seedlings by regulating ion homeostasis

Appropriate amounts of polyamines (PAs) alleviating salt-induced damage have been widely recognized. Maintaining ion homeostasis is one of the key mechanisms to improve salt tolerance. The present study aimed to explore the specific mechanism of putrescine (Put) regulating ion balance in NaCl-stressed cucumber (Cucumis sativus) seedlings. Biochemical analysis combined with microscope imaging techniques revealed that 0.8 mM Put effectively inhibited the accumulation of Na+ and the loss of K+ in cucumber plants exposed to 75 mM NaCl. More Na+ tended to accumulate in stem. Moreover, exogenous Put further increased K+ concentration but decreased Na+ concentration in xylem sap of cucumber seedlings stressed by NaCl. Put increasing expression levels of SOS1 and NHX was involved in the Na+ exclusion. The differential expression of HKT1 in roots and leaves caused by Put might regulate Na+ distribution. In addition, the application of N,N′-dimethylthiourea (DMTU) and aminoguanidine (AG) decreased H2O2 levels, accompanied by the increase of Na+ in Put-treated seedlings, suggesting that polyamines oxidation-generated H2O2 mediated the Put-regulated ion homeostasis to some extent. These results indicated that exogenous Put could interact with ion channels as well as induce the generation of H2O2 to maintain Na+/K+ homeostasis under NaCl stress conditions.

Polyamines and amino acids in triticale plants grown on humic acids enriched nutrient solution and treated with UV-B irradiation

The effects of UV-B irradiation and Biomin (a natural substance extracted from coal with active ingredients of humic acids) on the content of endogenous polyamines spermine, spermidine and putrescine, and free amino acids in shoots and roots of young triticale seedlings were investigated. Biomin was added to the nutrient medium 3 days prior to UV-B irradiation. The seedlings were treated with 7.7 kJ m−2 day−1 UV-B light for 4 days. The exposure to UV-B increased total free amino acids, while Biomin application alone did not affect considerably their content. The treatment with UV-B or Biomin alone provoked augmentation of conjugated and bound polyamine (PA) fractions. Data suggest that Biomin alleviates the negative consequences of UV-B stress, manifested by the normalized amino acid and polyamine amounts in the UV-B + Biomin-treated plants. This study demonstrates the protective effect of Biomin on triticale plants against UV-B irradiation, which could be related to alterations in PAs and amino acids pools.

Tuning Enzyme/α-Zr(IV) Phosphate Nanoplate Interactions via Chemical Modification of Glucose Oxidase.

Using glucose oxidase (GOx) and α-Zr(IV) phosphate nanoplates (α-ZrP) as a model system, a generally applicable approach to control enzyme-solid interactions via chemical modification of amino acid side chains of the enzyme is demonstrated. Net charge on GOx was systematically tuned by appending different amounts of polyamine to the protein surface to produce chemically modified GOx(n), where n is the net charge on the enzyme after the modification and ranged from -62 to +95 electrostatic units in the system. The binding of GOx(n) with α-ZrP nanosheets was studied by isothermal titration calorimetry (ITC) as well as by surface plasmon resonance (SPR) spectroscopy. Pristine GOx showed no affinity for the α-ZrP nanosheets, but GOx(n) where n ≥ -20 showed binding affinities exceeding (2.1 ± 0.6) × 106 M-1, resulting from the charge modification of the enzyme. A plot of GOx(n) charge vs Gibbs free energy of binding (ΔG) for n = +20 to n = +65 indicated an overall increase in favorable interaction between GOx(n) and α-ZrP nanosheets. However, ΔG is less dependent on the net charge for n > +45, as evidenced by the decrease in the slope as charge increased further. All modified enzyme samples and enzyme/α-ZrP complexes retained a significant amount of folding structure (examined by circular dichroism) as well as enzymatic activities. Thus, strong control over enzyme-nanosheet interactions via modulating the net charge of enzymes may find potential applications in biosensing and biocatalysis.

Study on improving plantlet development and embryo germination rates in in vitro embryo rescue of seedless grapevine

ABSTRACTThe embryo rescue technique has been proved to be highly effective and become a standard tool in seedless grape breeding. The main factors limiting the success of in vitro embryo rescue are the germination and development of zygotic embryos. Here, we evaluate the effects of different crossing parents, sampling time and culture medium, including different polyamines on development processes of stenospermic grape embryos. Our results indicate that both embryo germination and plantlet development were highly dependent on the crossing parent genotypes, sampling times and improvement of culture medium. Additions of 3 mM putrescine, 0.5 mM spermidine or 0.3 mM spermine significantly increase plantlet development or embryo germination rate in a range of hybrid combinations, including Sultanina Rose × Shuangyou, Ruby Seedless × Zixiang Seedless, Aishenmeigui × Shine-Muscat, Ruby Seedless × Flame Seedless and Blush Seedless × Aishenmeigui, which reveals that appropriate amounts of polyamines are added to the culture medium, the efficiency of in vitro embryo rescue can be significantly increased.

PbrMYB21, a novel MYB protein of Pyrus betulaefolia, functions in drought tolerance and modulates polyamine levels by regulating arginine decarboxylase gene.

Summary MYB comprises a large family of transcription factors that play significant roles in plant development and stress response in plants. However, knowledge concerning the functions of MYBs and the target genes remains poorly understood. Here, we report the identification and functional characterization of a novel stress‐responsive MYB gene from Pyrus betulaefolia. The MYB gene, designated as PbrMYB21, belongs to the R2R3‐type and shares high degree of sequence similarity to MdMYB21. The transcript levels of PbrMYB21 were up‐regulated under various abiotic stresses, particularly dehydration. PbrMYB21 was localized in the nucleus with transactivation activity. Overexpression of PbrMYB21 in tobacco conferred enhanced tolerance to dehydration and drought stresses, whereas down‐regulation of PbrMYB21 in Pyrus betulaefolia by virus‐induced gene silencing (VIGS) resulted in elevated drought sensitivity. Transgenic tobacco exhibited higher expression levels of ADC (arginine decarboxylase) and accumulated larger amount of polyamine in comparison with wild type (WT). VIGS of PbrMYB21 in Pyrus betulaefolia down‐regulated ADC abundance and decreased polyamine level, accompanied by compromised drought tolerance. The promoter region of PbrADC contains one MYB‐recognizing cis‐element, which was shown to be interacted with PbrMYB21, indicating the ADC may be a target gene of PbrMYB21. Take together, these results demonstrated that PbrMYB21 plays a positive role in drought tolerance, which may be, at least in part, due to the modulation of polyamine synthesis by regulating the ADC expression.

Variation in Polyamine Content among 12 Pollinated Wheat Genotypes under Drought Stress Condition

The present work aims to study the drought stress effects on polyamine content and its relationship with growth and development in seeds, during cell division (from the beginning and until 17 days after pollination) and grain filling (until reaching the physiological maturity) stages. Factorial experiment based on Randomized Complete Block (RCB) design was carried out with three replications. Two factors of the experiment were considered: the level of irrigation (irrigation without and with drought stress during cell division stage and with drought stress during grain filling stage) and the level of wheat genotype (drought-resistant, semi-resistant and sensitive genotypes). As control treatment, soil moisture content of the field was considered. For drought-stress treatment, the 50% of the soil moisture content in control treatment was established. According to the experimental results, Putrescine content value is higher in control treatment which develops seeds of drought-resistant genotypes than that are registered for semi-resistant and sensitive genotypes. Both drought stress treatments induced significant rises in putrescine amount in the different genotypes of the study. The rises of putrescine content in sensitive and semi-resistant genotypes, however, were higher than in resistant ones, and the highest rise in putrescine content appeared in manning semi-resistant genotype during the stage of grain filling (27 days after pollination). Exerting drought stress in both stages of cell division and grain filling led to significant increase in spermine and spermidine contents of developing seeds of entire genotypes of the study. The highest spermidine content during grain filling stage occurred in sensitive genotypes of Shark and Tevee’s, and the lowest occurred in semi-resistant genotype of Sabalan. The highest spermine content during cell division stage, though, happened in semi-resistant genotype of manning. In fact, spermine and spermidine act as analogous to plant hormones such as Auxin and Cytokine, or they cooperate with these hormones in order to take a role in cell division regulation of developing wheat seeds and development of sink strength. They, additionally, stabilize the cell division process of developing seeds under drought condition. According to the findings of the study, manning semi-resistant genotype is capable of having a high grain yield under drought condition by reason of reserving high amounts of polyamines.

Screening and Quantification of Aliphatic Primary Alkyl Corrosion Inhibitor Amines in Water Samples by Paper Spray Mass Spectrometry.

Direct analysis and identification of long chain aliphatic primary diamine Duomeen O (n-oleyl-1,3-diaminopropane), corrosion inhibitor in raw water samples taken from a large medium pressure water tube boiler plant water samples at low LODs (<0.1 pg) has been demonstrated for the first time, without any sample preparation using paper spray mass spectrometry (PS-MS). The presence of Duomeen O in water samples was confirmed via tandem mass spectrometry using collision-induced dissociation and supported by exact mass measurement and reactive paper spray experiments using an LTQ Orbitrap Exactive instrument. Data shown herein indicate that paper spray ambient ionization can be readily used as a rapid and robust method for in situ direct analysis of polymanine corrosion inhibitors in an industrial water boiler plant and other related samples in the water treatment industry. This approach was applied for the analysis of three complex water samples including feedwater, condensate water, and boiler water, all collected from large medium pressure (MP) water tube boiler plants, known to be dosed with varying amounts of polyamine and amine corrosion inhibitor components. Polyamine chemistry is widely used for example in large high pressure (HP) boilers operating in municipal waste and recycling facilities to prevent corrosion of metals. The samples used in this study are from such a facility in Coventry waste treatment facility, U.K., which has 3 × 40 tonne/hour boilers operating at 17.5 bar.

Вміст амінокислот і поліамінів у грудному м’язі курчат-бройлерів за умов застосування екстракту селезінки з метою нівелювання передзабійного стресу

The results on the Adrenocorticotropic hormone (ACTH) level in blood, amino acids and polyamines content in pectoral muscle of broiler chickens under using spleen extract for decreasing pre-slaughter stress are presented. The spleen extract (70% alcohol solution in volume 1.4 ml per chicken) was added to the diet of broiler chi­ckens of experimental groups by using aerosol method. This extract was obtained with and without the ultrasound application. 70% alcohol solution in the same volume and the same method was used to the diet of broiler chickens of control group five days before slaughter. The content of polyamines, such as putrescin, spermine and spermidin in pectoral muscle of broiler chi­ckens was determined. The total amount of polyamines and spermine specifically of both experimental groups in broiler chickens pectoral muscle was reliable higher compared to the control. The level of putrescin, spermine and spermidin was the lowest in pectoral muscle of broiler chickens of control group. The putrescin content in the pectoral muscle was reliablely highest in broiler chickens of the first experimental group after addition to the diet of the spleen extract with the ultrasound application. The concentration of amino acids, except of serine, in broiler chickens pectoral muscle was reliablely highest in broiler chickens of both experimental group, compared to control. The amount of polyamines and spermine specifically of both experimental groups in pectoral muscle of broi­ler chickens was reliablely higher, compared to control. After using of spleen extract, obtained with and without the ultrasound application, ACTH level in blood of broiler chi­ckens was reliablely lower compared to control, that caused a decrease at the pre-slaughter stress. The amino acids content in pectoral muscle of broiler chickens was reliablely the highest in broiler chickens of both experimental group after addition of the spleen extract to the diet that caused a raise in biological quality of poultry meat.

Inhibition of Polyamine Formation Antagonizes Vascular Smooth Muscle Cell Proliferation and Preserves the Contractile Phenotype

The polyamines putrescine, spermidine and spermine play essential roles in cell proliferation and migration, two processes involved in the development of vascular disease. Thus, intervention with polyamine formation may represent a way to inhibit unwanted vascular smooth muscle cell (VSMC) proliferation. The aim of the present study was to assess the importance of polyamines for VSMC proliferation and vascular contractility. The rate-limiting step in polyamine biosynthesis is catalysed by ornithine decarboxylase (ODC). Treatment with α-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, reduced DNA synthesis in primary rat VSMCs in a concentration-dependent manner with an IC50 value of 100μM. Moreover, DFMO reduced VSMC migration assessed in a scratch assay. The DFMO-induced attenuation of VSMC proliferation was associated with lowered cellular amount of polyamines. The antiproliferative effect of DFMO was specific because supplementation with polyamines reversed the effect of DFMO on proliferation and normalized cellular polyamine levels. Isometric force recordings in cultured rat tail artery rings showed that DFMO counteracts the decrease in contractility caused by culture with foetal bovine serum as growth stimulant. We conclude that inhibition of polyamine synthesis by DFMO may limit the first wave of cell proliferation and migration, which occurs in the acute phase after vascular injury. Besides its antiproliferative effect, DFMO may prevent loss of the smooth muscle contractile phenotype in vascular injury.

Polyamine supplementation in infant formula: Influence on lymphocyte populations and immune system-related gene expression in a Balb/cOlaHsd mouse model

The aim of this work was to study whether the proportion of polyamine found in human milk, administered with a commercial infant formula, affected the maturation of the immune system in a BALB/cOlaHsd mouse model. Forty-eight pups (14-days old) were randomly assigned to four-day intervention groups: 1) breast-fed (normal lactation); 2) fed infant formula; and 3) two different groups fed with infant formula supplemented with two different amounts of polyamines. The influence of polyamine administration on lymphocyte populations in the blood, spleen, and mesenteric lymph nodes, as well as on the modulation of immune system-related gene expression in the small intestine, was analyzed. The results demonstrated that polyamine supplementation induced an increase in splenic B cells to levels observed during normal lactation when compared with formula without supplementation. The correlation coefficients for the splenic lymphocyte populations increased with polyamine supplementation, with a dose-dependent effect. Our results demonstrate that polyamines influence gene expression profile, mainly Cd1d1, Cd40, Hdac5, Hdac7, Clcf1 and Tlr4 compared with normal lactation. In general, the gene expression results verified that the expression of genes associated with immune system was similar in the group with high polyamine supplementation to that observed in the group with normal lactation.