RESUMEN

Researchers have reported the benefits of feeding rumen-protected methionine (RPM) during the peripartum on the health parameters of dairy cows. Rumen-protected Met has reportedly improved milk yield, milk components, and liver health, but the literature is scarce on its effects in commercial herds. Therefore, we aimed to determine the effects of feeding RPM (Smartamine M, Adisseo Inc., Antony, France) prepartum (8 g/cow per day) and postpartum (15 g/cow per day) on performance, metabolic profile, and culling rate of Holstein cows in a commercial herd. One hundred sixty-six (n = 166) Holstein cows, 58 nulliparous and 108 parous, were randomly assigned to 1 of 2 dietary treatments, consisting of TMR top-dressed with RPM (RPMet; 2.35% and 2.24% Met of MP for close-up and fresh cows, respectively) or without (control [CON] 2.03% and 1.89% Met of MP for close-up and fresh cows, respectively), fed from 21 ± 6 d prepartum until 16 ± 5 d postpartum. From 17 DIM until dry-off, all cows received RPMet. Daily milk yield was recorded, and milk samples were collected in the first and second weeks after calving to determine their composition. Blood samples were collected before the morning feeding on -14, -7, +1, +7, and +14 d relative to calving. Mortality and morbidity were recorded during the first 60 DIM. Cows supplemented with RPMet had greater milk yield during the first 16 DIM (31.76 vs. 30.37 kg/d; SEM = 1.04, respectively), and had greater milk fat content (4.45 vs. 4.10%; SEM = 0.11, respectively), but not milk total protein (3.47 vs. 3.39%; SEM = 0.04, respectively) and casein contents (2.74 vs. 2.66%; SEM = 0.04, respectively) than CON cows. Cows in RPMet had increased plasma Met concentrations than cows in CON (24.9 vs. 21.0 µmol/L; SEM = 1.2, respectively). Although morbidity was similar between treatments, the culling rate from calving until 60 DIM was lower for RPMet cows than for CON cows (2.4% vs. 12.1%; SEM = 0.02). In conclusion, cows receiving RPMet have greater milk yield, improved milk fat content, and a lower culling rate at 60 DIM than CON cows.

Asunto(s)

Alimentación Animal , Dieta , Lactancia , Metionina , Leche , Periodo Periparto , Rumen , Animales , Bovinos , Metionina/metabolismo , Metionina/administración & dosificación , Femenino , Leche/química , Leche/metabolismo , Dieta/veterinaria , Rumen/metabolismo , Alimentación Animal/análisis

RESUMEN

The oxidation of Met to methionine sulfoxide (MetSO) by oxidants such as hydrogen peroxide, hypochlorite, or peroxynitrite has profound effects on protein function. This modification can be reversed by methionine sulfoxide reductases (msr). In the context of pathogen infection, the reduction of oxidized proteins gains significance due to microbial oxidative damage generated by the immune system. For example, Mycobacterium tuberculosis (Mt) utilizes msrs (MtmsrA and MtmsrB) as part of the repair response to the host-induced oxidative stress. The absence of these enzymes makes Mycobacteria prone to increased susceptibility to cell death, pointing them out as potential therapeutic targets. This study provides a detailed characterization of the catalytic mechanism of MtmsrA using a comprehensive approach, including experimental techniques and theoretical methodologies. Confirming a ping-pong type enzymatic mechanism, we elucidate the catalytic parameters for sulfoxide and thioredoxin substrates (kcat/KM = 2656 ± 525 M-1 s-1 and 1.7 ± 0.8 × 106 M-1 s-1, respectively). Notably, the entropic nature of the activation process thermodynamics, representing ∼85% of the activation free energy at room temperature, is underscored. Furthermore, the current study questions the plausibility of a sulfurane intermediate, which may be a transition-state-like structure, suggesting the involvement of a conserved histidine residue as an acid-base catalyst in the MetSO reduction mechanism. This mechanistic insight not only advances our understanding of Mt antioxidant enzymes but also holds implications for future drug discovery and biotechnological applications.

Asunto(s)

Metionina Sulfóxido Reductasas , Mycobacterium tuberculosis , Metionina Sulfóxido Reductasas/metabolismo , Mycobacterium tuberculosis/metabolismo , Oxidación-Reducción , Catálisis , Estrés Oxidativo , Metionina/metabolismo

RESUMEN

Rickettsia typhi is the causative agent of murine typhus (endemic typhus), a febrile illness that can be self-contained, though in some cases it can progress to death. The three dimensional structure of Methionyl-tRNA Synthetase from R. typhi (RtMetRS) in complex with its substrate l-methionine was solved by molecular replacement and refined at 2.30 Å resolution in space group P1 from one X-ray diffraction dataset. Processing and refinement trials were decisive to establish the lower symmetry space group and indicated the presence of twinning with four domains. RtMetRS belongs to the MetRS1 family and was crystallized with the CP domain in an open conformation, what is distinctive from other MetRS1 enzymes whose structures were solved with a bound L-methionine (therefore, in a closed conformation). This conformation resembles the ones observed in the MetRS2 family.

Asunto(s)

Metionina-ARNt Ligasa , Animales , Ratones , Metionina-ARNt Ligasa/química , Metionina-ARNt Ligasa/genética , Metionina-ARNt Ligasa/metabolismo , Aminoácidos , Rickettsia typhi/metabolismo , Difracción de Rayos X , Metionina/metabolismo

RESUMEN

Tambaqui (Colossoma macropomum) is a species of great cultural and economic importance in aquaculture in the Amazon region. Methionine is considered the first limiting sulfur amino acid in practical fish diets, which encourages investigating its use in diets for tambaqui. This study aimed to verify the digestible methionine plus cystine (Met + Cys) requirement in diets for tambaqui (89.52 ± 0.53 g) for 60 days. The treatments investigated were: 6.50, 7.80, 9.10, 10.40, 11.70, and 13.00 g Met + Cys kg diet-1. The estimated requirement based on final weight, weight gain, feed conversion ratio, and specific growth rate was 9.04, 8.92, 8.91, and 8.58 g Met + Cys kg diet-1, respectively, while on body protein deposition, body fat deposition, body ash deposition, and nitrogen retention efficiency was 9.29, 9.20, 9.19, and 8.72 g Met + Cys kg diet-1, respectively. Linear regression demonstrated that increased digestible Met + Cys in the diet decreased plasma total protein, globulin, and liver total protein levels. Quadratic regression showed that the highest value for liver glycogen was found with a 10.40 g Met + Cys kg diet-1. Another quadratic regression demonstrated a lower hepatic aspartate aminotransferase (AST) enzymatic activity in fish fed between 7.80 and 11.70 g Met + Cys kg diet-1. The different treatments did not influence the erythrogram. In conclusion, when considering an integrative view of the results for growth performance, whole-body deposition, and liver parameters without harming the physiological and metabolic status, we recommended choosing a diet with digestible Met + Cys between 8.58 and 9.29 g kg- 1 for tambaqui.

Asunto(s)

Aminoácidos Sulfúricos , Metionina , Animales , Metionina/metabolismo , Cistina/metabolismo , Aminoácidos Sulfúricos/metabolismo , Racemetionina/metabolismo , Dieta/veterinaria , Composición Corporal , Hígado/metabolismo , Alimentación Animal/análisis

RESUMEN

Trypanosoma cruzi is the causal agent of Chagas Disease and is a unicellular parasite that infects a wide variety of mammalian hosts. The parasite exhibits auxotrophy by L-Met; consequently, it must be acquired from the extracellular environment of the host, either mammalian or invertebrate. Methionine (Met) oxidation produces a racemic mixture (R and S forms) of methionine sulfoxide (MetSO). Reduction of L-MetSO (free or protein-bound) to L-Met is catalyzed by methionine sulfoxide reductases (MSRs). Bioinformatics analyses identified the coding sequence for a free-R-MSR (fRMSR) enzyme in the genome of T. cruzi Dm28c. Structurally, this enzyme is a modular protein with a putative N-terminal GAF domain linked to a C-terminal TIP41 motif. We performed detailed biochemical and kinetic characterization of the GAF domain of fRMSR in combination with mutant versions of specific cysteine residues, namely, Cys12, Cys98, Cys108, and Cys132. The isolated recombinant GAF domain and full-length fRMSR exhibited specific catalytic activity for the reduction of free L-Met(R)SO (non-protein bound), using tryparedoxins as reducing partners. We demonstrated that this process involves two Cys residues, Cys98 and Cys132. Cys132 is the essential catalytic residue on which a sulfenic acid intermediate is formed. Cys98 is the resolutive Cys, which forms a disulfide bond with Cys132 as a catalytic step. Overall, our results provide new insights into redox metabolism in T. cruzi, contributing to previous knowledge of L-Met metabolism in this parasite.

Asunto(s)

Metionina Sulfóxido Reductasas , Trypanosoma cruzi , Metionina Sulfóxido Reductasas/genética , Metionina Sulfóxido Reductasas/química , Metionina Sulfóxido Reductasas/metabolismo , Trypanosoma cruzi/genética , Oxidación-Reducción , Cisteína/química , Metionina/metabolismo

RESUMEN

Supplementation of rumen-protected amino acids may improve dairy cow performance but few studies have evaluated the implications of supplementing low-forage diets. Our objective was to evaluate the effects of supplementing rumen-protected methionine (Met) and lysine (Lys) on milk production and composition as well as on mammary gland health of mid-lactating Holstein cows from a commercial dairy farm feeding a high by-product low-forage diet. A total of 314 multiparous cows were randomly assigned to control (CON; 107 g of dry distillers' grains) or rumen-protected Met and Lys (RPML; 107 g dry distillers' grains + 107 g of RPML). All study cows were grouped in a single dry-lot pen and fed the same total mixed ration diet twice a day for a total of 7 weeks. Treatments were top-dressed on the total mix ration immediately after morning delivery with 107 g of dry distillers' grains for 1 week (adaptation period) and then with CON and RPML treatments for 6 weeks. Blood samples were taken from a subset of 22 cows per treatment to determine plasma AA (d 0 and 14) and plasma urea nitrogen and minerals (d 0, 14, and 42). Milk yield and clinical mastitis cases were recorded daily, and milk components were determined bi-weekly. Body condition score change was evaluated from d 0 to 42 of the study. Milk yield and components were analyzed by multiple linear regression. Treatment effects were evaluated at the cow level considering parity and milk yield and composition taken at baseline as a covariate in the models. Clinical mastitis risk was assessed by Poisson regression. Plasma Met increased (26.9 vs 36.0 µmol/L), Lys tended to increase (102.5 vs 121.1 µmol/L), and Ca increased (2.39 vs 2.46 mmol/L) with RPML supplementation. Cows supplemented with RPML had higher milk yield (45.4 vs 46.0 kg/d) and a lower risk of clinical mastitis (risk ratio = 0.39; 95% CI = 0.17-0.90) compared to CON cows. Milk components yield and concentrations, somatic cell count, body condition score change, plasma urea nitrogen, and plasma minerals other than Ca were not affected by RPML supplementation. Results suggest that RPML supplementation increases milk yield and decreases the risk of clinical mastitis in mid-lactation cows fed a high by-product low-forage diet. Further studies are needed to clarify the biological mechanisms for mammary gland responses to RPML supplementation.

Asunto(s)

Lactancia , Lisina , Embarazo , Femenino , Bovinos , Animales , Lisina/metabolismo , Lactancia/fisiología , Metionina/metabolismo , Paridad , Rumen/metabolismo , Dieta/veterinaria , Suplementos Dietéticos , Leche/metabolismo , Racemetionina/metabolismo , Nitrógeno/metabolismo , Minerales/metabolismo , Urea/metabolismo , Alimentación Animal/análisis

RESUMEN

This study investigated the hypothesis that methionine supplementation of Japanese quail (Coturnix coturnix japonica) hens can reduce the effects of oxidative stress and improve the performance of the offspring exposed to heat stress during growth. For that, the quail hens were fed with three diets related to the methionine supplementation: methionine-deficient diet (Md); diet supplemented with the recommended methionine level (Met1); and diet supplemented with methionine above the recommended level (Met2). Their chicks were identified, weighed, and housed according to the maternal diet group from 1 to 14 d of age. On 15 d of age, chicks were weighed and divided into two groups: thermoneutral ambient (constant temperature of 23 °C) and intermittent heat stress ambient (daily exposure to 34 °C for 6 h). Methionine-supplemented (Met1 and Met2) hens had higher egg production, better feed conversion ratio, higher hatchability of total and fertile eggs, and offspring with higher body weight. Supplemented (Met1 and Met2) hens showed greater expression of glutathione synthase (GSS) and methionine sulfoxide reductase A (MSRA) genes, greater total antioxidant capacity, and lower lipid peroxidation in the liver. The offspring of hens fed the Met2 diet had lower death rate (1 to 14 d), higher weight on 15 d of age, weight gain, and better feed conversion ratio from 1 to 14 d of age. Among chicks reared under heat stress, the progeny of methionine-supplemented hens had higher weight on 35 d, weight gain, expression of GSS, MSRA, and thermal shock protein 70 (HSP70) genes, and total antioxidant capacity in the liver, as well as lower heterophil/lymphocyte ratio. Positive correlations between expression of glutathione peroxidase 7 (GPX7) and MSRA genes in hens and offspring were observed. Our results show that maternal methionine supplementation contributes to offspring development and performance in early stages and that, under conditions of heat stress during growth, chicks from methionine-supplemented hens respond better to hot environmental conditions than chicks from nonsupplemented hens. Supplementation of quail hens diets with methionine promoted activation of different metabolic pathways in offspring subjected to stress conditions.

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The deficiency of nutrients such as methionine in the diet of birds is affecting fertility rate, egg production, egg weight, and progeny weight. In addition, the maternal environment influences gene expression through epigenetic mechanisms, where the conditions experienced by the parental generation during embryonic development can produce effects on the progeny. This study investigates how methionine supplementation in the diet of quail hens can reduce the effects of oxidative stress and improve the performance of progeny subjected to heat stress during growth. For that, the quail hens were fed with diets containing three different levels of methionine; and their chicks were created (15 on 35 d of age) into thermoneutral and/or intermittent heat stress ambient. It was observed that methionine supplementation in the quail hens had a positive effect on mortality during the initial phase and greater weight gain in the progeny growth phase. In addition, genetic inheritance was observed through the positive correlation between the expression of genes (maternal and progeny) related to oxidative stress. The results show that methionine supplementation in the maternal diet contributes to the development and performance of the progeny when subjected to heat stress during the growth phase.

Asunto(s)

Antioxidantes , Coturnix , Animales , Femenino , Antioxidantes/metabolismo , Coturnix/fisiología , Dieta/veterinaria , Suplementos Dietéticos , Respuesta al Choque Térmico , Metionina/farmacología , Metionina/metabolismo , Óvulo , Codorniz , Racemetionina/metabolismo , Aumento de Peso

RESUMEN

Nonalcoholic steatohepatitis (NASH) is a disease with a high incidence worldwide, but its diagnosis and treatment are poorly managed. In this study, NASH pathophysiology and DNA damage biomarkers were investigated in mice with NASH treated and untreated with melatonin (MLT). C57BL/6 mice were fed a methionine- and choline-deficient (MCD) diet for 4 weeks to develop NASH. Melatonin was administered at 20 mg/kg during the last 2 weeks. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were measured, and hepatic tissue was dissected for histological analysis, evaluation of lipoperoxidation, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), as well as nuclear factor-erythroid 2 (Nrf2), tumor necrosis factor alpha (TNF-α), inducible nitric oxide synthase (iNOS), and transforming growth factor beta (TGF-ß) expression by immunohistochemistry. DNA damage was evaluated using Comet assay, while a micronucleus test in bone marrow was performed to assess the genomic instability associated with the disease. Melatonin decreased AST and ALT, liver inflammatory processes, balloonization, and fibrosis in mice with NASH, decreasing TNF-α, iNOS, and TGF-ß, as well as oxidative stress, shown by reducing lipoperoxidation and intensifying Nrf2 expression. The SOD and GPx activities were increased, while CAT was decreased by treatment with MLT. Although the micronucleus frequency was not increased in mice with NASH, a protective effect on DNA was observed with MLT treatment in blood and liver tissues using Comet assay. As conclusions, MLT slows down the progression of NASH, reducing hepatic oxidative stress and inflammatory processes, inhibiting DNA damage via anti-inflammatory and antioxidant actions.

Asunto(s)

Deficiencia de Colina , Melatonina , Enfermedad del Hígado Graso no Alcohólico , Alanina Transaminasa , Animales , Antiinflamatorios/farmacología , Antioxidantes/metabolismo , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Aspartato Aminotransferasas , Biomarcadores/metabolismo , Catalasa/metabolismo , Colina/análisis , Colina/metabolismo , Colina/farmacología , Deficiencia de Colina/complicaciones , Deficiencia de Colina/metabolismo , Daño del ADN , Dieta , Glutatión Peroxidasa/metabolismo , Inflamación/metabolismo , Hígado/metabolismo , Melatonina/farmacología , Melatonina/uso terapéutico , Metionina/análisis , Metionina/genética , Metionina/metabolismo , Ratones , Ratones Endogámicos C57BL , Factor 2 Relacionado con NF-E2/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Estrés Oxidativo , Superóxido Dismutasa/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

1. Cobb and Ross broilers (200 of each sex and breed) were fed four phases of diets ad libitum formulated with balanced protein to match their amino acid requirements throughout growth. Ten birds per genotype were sampled and euthanised at two-weekly intervals from 14 to 112 d of age. All feathers were dry-plucked from each of the seven tracts (specific skin areas) and pulp (the centre of the feather filament) was removed from primary and secondary remiges.2. Daily losses of feathers were collected from an additional 20 individually-caged broilers of each breed. These feathers were separated into natal down, contour feathers, remiges and rectrices and then pooled by type, sex and genotype to quantify water and protein contents. Only those feathers collected from male Cobb 500 MX broilers were analysed for amino acid content.3. Amino acid contents of feathers from the seven tracts were measured only in Cobb males on days 1, 28 and 70; for pulp on days 28 and 70; and for the four types of moulted feathers.4. Protein content on a dry matter basis remained relatively constant over all ages and tracts during growth. Water content decreased with age in both sexes and genotype. Lysine and methionine content in feathers decreased with age while cystine, valine, leucine and serine increased. Lysine, methionine and histidine levels were higher in pulp than in mature feathers whereas cystine and valine were higher in mature feathers than in pulp.5. These results, together with information about moulting patterns in broilers, enabled the effects of age of the bird and of the type of feather, to be taken into account when determining the rate of deposition of amino acids in feathers.

Asunto(s)

Pollos , Plumas , Aminoácidos/metabolismo , Animales , Cistina/metabolismo , Plumas/química , Femenino , Genotipo , Lisina/análisis , Masculino , Metionina/metabolismo , Proteínas/análisis , Valina/análisis , Valina/metabolismo , Agua/análisis

RESUMEN

Amyloid aggregation of α-synuclein (AS) is one of the hallmarks of Parkinson's disease (PD). Copper ions specifically bind at the N-terminus of AS, accelerating protein aggregation. Its protein homolog ß-synuclein (BS) is also a copper binding protein, but it inhibits AS aggregation. Here, a comparative spectroscopic study of the Cu2+ binding properties of AS and BS has been performed, using electronic absorption, circular dichroism (CD) and electronic paramagnetic resonance (EPR). Our comparative spectroscopic study reveals striking similarities between the Cu2+ binding features of the two proteins. The Cu2+ binding site at the N-terminal group of BS protein, modeled by the BS (1-15) fragment is identical to that of AS; however, its rate of reduction is three times faster as compared to the AS site, consistent with BS having an additional Met residue in its Met1-Xn-Met5-Xn-Met10 motif. The latter is also evident in the cyclic voltammetry studies of the Cu-BS complex. On the other hand, the Cu2+ binding features of the His site in both proteins, as modeled by AS(45-55) and BS(60-70), are identical, indicating that the shift in the His position does not affect its coordination features. Finally, replacement of Glu46 by Ala does not alter Cu2+ binding to the His site, suggesting that the familial PD E46K mutation would not impact copper-induced aggregation. While further studies of the redox activity of copper bound to His50 in AS are required to understand the role of this site in metal-mediated aggregation, our study contributes to a better understanding of the bioinorganic chemistry of PD.

Asunto(s)

Cobre/metabolismo , alfa-Sinucleína/metabolismo , Sinucleína beta/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Histidina/química , Histidina/metabolismo , Metionina/química , Metionina/metabolismo , Unión Proteica , alfa-Sinucleína/química , Sinucleína beta/química

RESUMEN

Increased homocysteine (Hcy) levels have been associated with a higher risk of cardiovascular and neurodegenerative diseases. Passive DNA demethylation has been suggested as one of the mechanisms implicated in the development of these conditions, and most studies have investigated this relationship in older adult populations. Therefore, this study aimed to evaluate the relationship between corporal composition and biochemical and haematological indicators with plasma homocysteine levels and genome-wide methylation (Alu, LINE-1, and SAT2) in a population of healthy young adults (median age, 18 years). We showed that the prevalence of hyperhomocysteinemia was significantly higher in men (18.5%) than in women (6.6%) (P = 0.034). Increased Hcy level was substantially associated with higher levels of body mass index and visceral fat in females, whereas in males, it was significantly associated with reduced red cell distribution width and high-density lipoprotein (HDL) cholesterol (HDL-C) levels and increased low-density lipoprotein/HDL ratio. Hypomethylation of Alu was significantly associated with reduced levels of HDL-C (<40.0 mg dL-1), whereas hypomethylation of LINE-1 and SAT2 was significantly associated with higher levels of skeletal muscle (<39.3%) in males. These results highlight the participation of hormonal factors in regulating Hcy metabolism, primarily in the female population, whereas changes in DNA methylation observed in males might be associated with the consumption of a protein diet with high levels of methionine, independent of increased Hcy levels.

Asunto(s)

Hiperhomocisteinemia , Adolescente , Anciano , HDL-Colesterol , Metilación de ADN , Femenino , Homocisteína/metabolismo , Humanos , Hiperhomocisteinemia/epidemiología , Hiperhomocisteinemia/genética , Lipoproteínas LDL/metabolismo , Masculino , Metionina/metabolismo , Adulto Joven

RESUMEN

Increasing methionine availability in dairy cow diets during the first third of lactation may enhance their performance and health. The aim of this study was to determine the effect of supplementing rumen-protected methionine (Smartamine® M, SM) in a lactation diet with protein and energy levels calculated according to the literature. Seventy-six multiparous Holstein cows (39.1 ± 6.8 kg of milk/d and 65 ± 28 DIM) were assigned to 1 of 2 dietary treatments (38/treatment) according to a randomized complete block design with a 2-wk (covariate) and 10-wk experimental period. Treatments were a basal diet (CON; 3.77 Lys:1Met); and CON + 23 g SM (2.97 Lys:1 Met). Individual milk samples were taken every 2 weeks to determine milk composition. Blood was collected from 24 cows on d+30 d to measure plasma AA levels. Body weight and body condition score (BCS) were measured at the beginning and the end of the experiment. The SM diet promoted higher milk yield (41.7 vs. 40.1 kg/d; P = 0.03). Energy-corrected milk yield (41.0 vs. 38.0 kg/d), milk protein yield (1.30 vs. 1.18 kg/d), milk protein (3.14% vs. 2.97%) and casein (2.39% vs. 2.28%) were also different (P < 0.01) as well as milk fat yield (1.42 vs. 1.29 kg/d; P = 0.02). A trend (P = 0.06) for higher milk fat % (3.41% vs. 3.21%) was observed. Both diets resulted in similar body weight, but CON-fed cows tended (P = 0.08) to have higher BCS. Higher plasma methionine levels were determined with SM compared with CON (29.6 vs. 18.4 µM; P < 0.01), but lysine and histidine were not different. Dietary supplementation of RPM improved productive performance by increasing milk yield and milk components yields, suggesting better dietary AA utilization when Met levels are adjusted in Lys-adequate lactation diets.

Asunto(s)

Alimentación Animal , Bovinos/fisiología , Suplementos Dietéticos , Metionina/metabolismo , Leche/metabolismo , Alimentación Animal/análisis , Crianza de Animales Domésticos , Animales , Suplementos Dietéticos/análisis , Femenino , Lactancia , Metionina/administración & dosificación , Metionina/análisis , Leche/química , Proteínas de la Leche/análisis , Proteínas de la Leche/metabolismo , Rumen/fisiología

RESUMEN

Anthocyanin biosynthesis is one of the most studied pathways in plants due to the important ecological role played by these compounds and the potential health benefits of anthocyanin consumption. Given the interest in identifying new genetic factors underlying anthocyanin content we studied a diverse collection of diploid potatoes by combining a genome-wide association study and pathway-based analyses. By using an expanded SNP dataset, we identified candidate genes that had not been associated with anthocyanin variation in potatoes, namely a Myb transcription factor, a Leucoanthocyanidin dioxygenase gene and a vacuolar membrane protein. Importantly, a genomic region in chromosome 10 harbored the SNPs with strongest associations with anthocyanin content in GWAS. Some of these SNPs were associated with multiple anthocyanin compounds and therefore could underline the existence of pleiotropic genes or anthocyanin biosynthetic clusters. We identified multiple anthocyanin homologs in this genomic region, including four transcription factors and five enzymes that could be governing anthocyanin variation. For instance, a SNP linked to the phenylalanine ammonia-lyase gene, encoding the first enzyme in the phenylpropanoid biosynthetic pathway, was associated with all of the five anthocyanins measured. Finally, we combined a pathway analysis and GWAS of other agronomic traits to identify pathways related to anthocyanin biosynthesis in potatoes. We found that methionine metabolism and the production of sugars and hydroxycinnamic acids are genetically correlated to anthocyanin biosynthesis. The results contribute to the understanding of anthocyanins regulation in potatoes and can be used in future breeding programs focused on nutraceutical food.

Asunto(s)

Antocianinas/biosíntesis , Vías Biosintéticas , Estudio de Asociación del Genoma Completo/métodos , Polimorfismo de Nucleótido Simple , Solanum tuberosum/metabolismo , Biología Computacional/métodos , Ácidos Cumáricos/metabolismo , Diploidia , Regulación de la Expresión Génica de las Plantas , Metionina/metabolismo , Fenilanina Amoníaco-Liasa/genética , Proteínas de Plantas/genética , Sitios de Carácter Cuantitativo , Solanum tuberosum/genética

RESUMEN

Our previous studies have shown that methionine supplementation could help to attenuate the effects of heat stress on the metabolism of broiler chickens. Here we investigated for the first time the effects of methionine supplementation in the form of DL-methionyl-DL-methionine on broilers subjected to heat stress during the growth phase. Broilers were divided into two groups; one group was reared under thermoneutral conditions and the other under continuous heat stress (30 ±â€¯1 °C, 60% relative humidity). Both groups were subdivided into three dietary treatments: a methionine-deficient (MD) diet, a diet supplemented with free methionine (DL-M), and a diet supplemented with methionine dipeptide (DL-MM). Broilers raised under chronic heat stress had lower feed intake and weight gain than broilers raised under thermoneutral conditions (P < 0.05). There were no differences in animal performance between methionine-supplemented diets (DL-M and DL-MM). Heat-stressed birds had significantly higher heterophil/lymphocyte (H/L) ratio than thermoneutral birds. Under heat stress, broilers fed DL-M and DL-MM diets had lower H/L ratio than birds fed the MD diet. Higher concentrations of carbonylated proteins and lower concentration of reduced glutathione were observed in broilers raised under heat stress. In comparing heat-stressed broilers, we found that birds fed the DL-M diet had lower concentrations of thiobarbituric acid-reactive substances and carbonylated proteins than those fed the MD diet (P < 0.05). Higher expression of glutathione peroxidase (GPX) and glutathione synthetase (GSS) genes was observed in heat-stressed broilers (P < 0.05). Under heat stress, the MD diet increased GPX expression compared with other diets. Under thermoneutral conditions, the DL-M diet resulted in the highest GSS expression. There was a negative correlation between DNA methylation and GPX and GSS expression. Our results showed that supplementation of broiler diets with free methionine or methionine dipeptide may help attenuate the effects of heat stress through enhanced activation of genes related to the glutathione antioxidant system. Methionine effects were found for gene regulation, gene expression, and post-translational processing.

Asunto(s)

Pollos , Metionina , Alimentación Animal/análisis , Animales , Pollos/metabolismo , Dieta/veterinaria , Suplementos Dietéticos/análisis , Dipéptidos , Calor , Metionina/metabolismo , Estrés Oxidativo , Temperatura

RESUMEN

Here, we present the draft genome sequence of Pseudomonas sp. GC01, a cadmium-resistant Antarctic bacterium capable of biosynthesizing CdS fluorescent nanoparticles (quantum dots, QDs) employing a unique mechanism involving the production of methanethiol (MeSH) from methionine (Met). To explore the molecular/metabolic components involved in QDs biosynthesis, we conducted a comparative genomic analysis, searching for the genes related to cadmium resistance and sulfur metabolic pathways. The genome of Pseudomonas sp. GC01 has a 4,706,645 bp size with a 58.61% G+C content. Pseudomonas sp. GC01 possesses five genes related to cadmium transport/resistance, with three P-type ATPases (cadA, zntA, and pbrA) involved in Cd-secretion that could contribute to the extracellular biosynthesis of CdS QDs. Furthermore, it exhibits genes involved in sulfate assimilation, cysteine/methionine synthesis, and volatile sulfur compounds catabolic pathways. Regarding MeSH production from Met, Pseudomonas sp. GC01 lacks the genes E4.4.1.11 and megL for MeSH generation. Interestingly, despite the absence of these genes, Pseudomonas sp. GC01 produces high levels of MeSH. This is probably associated with the metC gene that also produces MeSH from Met in bacteria. This work is the first report of the potential genes involved in Cd resistance, sulfur metabolism, and the process of MeSH-dependent CdS QDs bioproduction in Pseudomonas spp. strains.

Asunto(s)

Genoma Bacteriano , Genómica , Pseudomonas/genética , Pseudomonas/metabolismo , Adaptación Biológica/genética , Cadmio/metabolismo , Cadmio/toxicidad , Compuestos de Cadmio/química , Biología Computacional/métodos , Genómica/métodos , Redes y Vías Metabólicas , Metionina/metabolismo , Modelos Biológicos , Nanopartículas , Filogenia , Pseudomonas/clasificación , Sulfuros/química

RESUMEN

BACKGROUND: Methionine (Met) oxidation leads to a racemic mixture of R and S forms of methionine sulfoxide (MetSO). Methionine sulfoxide reductases (Msr) are enzymes that can reduce specifically each isomer of MetSO, both free and protein-bound. The Met oxidation could change the structure and function of many proteins, not only of those redox-related but also of others involved in different metabolic pathways. Until now, there is no information about the presence or function of Msrs enzymes in Leptospira interrogans. METHODS: We identified genes coding for putative MsrAs (A1 and A2) and MsrB in L. interrogans serovar Copenhageni strain Fiocruz L1-130 genome project. From these, we obtained the recombinant proteins and performed their functional characterization. RESULTS: The recombinant L. interrogans MsrB catalyzed the reduction of Met(R)SO using glutaredoxin and thioredoxin as reducing substrates and behaves like a 1-Cys Msr (without resolutive Cys residue). It was able to partially revert the in vitro HClO-dependent inactivation of L. interrogans catalase. Both recombinant MsrAs reduced Met(S)SO, being the recycle mediated by the thioredoxin system. LinMsrAs were more efficient than LinMsrB for free and protein-bound MetSO reduction. Besides, LinMsrAs are enzymes involving a Cys triad in their catalytic mechanism. LinMsrs showed a dual localization, both in cytoplasm and periplasm. CONCLUSIONS AND GENERAL SIGNIFICANCE: This article brings new knowledge about redox metabolism in L. interrogans. Our results support the occurrence of a metabolic pathway involved in the critical function of repairing oxidized macromolecules in this pathogen.

Asunto(s)

Citoplasma/química , Leptospira interrogans/genética , Metionina Sulfóxido Reductasas/genética , Metionina/metabolismo , Secuencia de Aminoácidos/genética , Catálisis , Citoplasma/enzimología , Genoma Bacteriano/genética , Humanos , Leptospira interrogans/enzimología , Metionina/química , Metionina/genética , Metionina Sulfóxido Reductasas/química , Metionina Sulfóxido Reductasas/ultraestructura , Oxidación-Reducción , Homología de Secuencia de Aminoácido , Estereoisomerismo , Especificidad por Sustrato

RESUMEN

The objective of this study was to estimate the sulphur amino acid (methionine + cystine) requirements and nitrogen endogenous losses in kittens aged 150 to 240 d. Thirty-six cats were distributed in six treatments (six cats per treatment) consisting of different concentrations of methionine + cystine (M + C): T1, 6.5 g/kg; T2, 8.8 g/kg; T3, 11.3 g/kg; T4, 13.6 g/kg; T5, 16.0 g/kg; and control, 6.5 g/kg. Diets were formulated by serial dilution of T5 (a diet relatively deficient in M + C but containing high protein concentrations) with a minimal nitrogen diet (MND). Thus, crude protein and amino acid concentrations in diets T1-T5 decreased by the same factor. The control diet was the T1 diet supplemented with adequate concentrations of M + C (6.5 g/kg; 8.8 g/kg; 11.3 g/kg; 13.6 g/kg and 16.0 g/kg). All diets were based on ingredients commonly used in extruded cat diets. Digestibility assays were performed for the determination of nitrogen balance. Nitrogen intake (NI) and nitrogen excretion (NEX) results data were fitted with an exponential equation to estimate nitrogen maintenance requirement (NMR), theoretical maximum for daily nitrogen retention (NRmaxT), and protein quality (b). M + C requirements were calculated from the limiting amino acid intake (LAAI) equation assuming a nitrogen retention of 45 to 65% NRmaxT. The NMR of kittens aged 150, 195, and 240 d was estimated at 595, 559, and 455 mg/kg body weight (BW)0.67 per day, respectively, and M + C requirements were estimated at 517, 664, and 301 mg/kg BW0.67 per day, respectively.

Asunto(s)

Gatos/metabolismo , Cistina/metabolismo , Metionina/metabolismo , Nitrógeno/metabolismo , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales/efectos de los fármacos , Animales , Cistina/administración & dosificación , Dieta/veterinaria , Suplementos Dietéticos/análisis , Femenino , Masculino , Metionina/administración & dosificación

RESUMEN

Hypermethioninemia is a disorder characterized by high plasma levels of methionine (Met) and its metabolites such as methionine sulfoxide (MetO). Studies have reported associated inflammatory complications, but the mechanisms involved in the pathophysiology of hypermethioninemia are still uncertain. The present study aims to evaluate the effect of chronic administration of Met and/or MetO on phenotypic characteristics of macrophages, in addition to oxidative stress, purinergic system, and inflammatory mediators in macrophages. In this study, Swiss male mice were subcutaneously injected with Met and MetO at concentrations of 0.35-1.2 g/kg body weight and 0.09-0.3 g/kg body weight, respectively, from the 10th-38th day post-birth, while the control group was treated with saline solution. The results revealed that Met and/or MetO induce an M1/classical activation phenotype associated with increased levels of tumor necrosis factor alpha and nitrite, and reduced arginase activity. It was also found that Met and/or MetO alter the activity of antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase, as well as the levels of thiol and reactive oxygen species in macrophages. The chronic administration of Met and/or MetO also promotes alteration in the hydrolysis of ATP and ADP, as indicated by the increased activity of ectonucleotidases. These results demonstrate that chronic administration of Met and/or MetO promotes activated pro-inflammatory profile by inducing M1/classical macrophage polarization. Thus, the changes in redox status and purinergic system upon chronic Met and/or MetO exposure may contribute towards better understanding of the alterations consistent with hypermethioninemic patients.

Asunto(s)

Errores Innatos del Metabolismo de los Aminoácidos/inmunología , Glicina N-Metiltransferasa/deficiencia , Macrófagos/inmunología , Metionina/análogos & derivados , Animales , Catalasa/metabolismo , Polaridad Celular , Glutatión Peroxidasa/metabolismo , Glicina N-Metiltransferasa/inmunología , Macrófagos/efectos de los fármacos , Masculino , Metionina/administración & dosificación , Metionina/metabolismo , Metionina/farmacología , Ratones , Oxidación-Reducción , Estrés Oxidativo , Fenotipo , Superóxido Dismutasa/metabolismo

RESUMEN

The anti-proliferative potential of Passiflora mollissima seeds, an underexplored agri-food waste, was investigated in this work by evaluating the molecular changes induced at transcript and metabolite expression levels on HT-29 human colon cancer cells. For this purpose, a pressurized-liquid extract from P. mollissima seeds obtained under optimized conditions was used for the treatment of HT-29 cells and a multi-omics strategy applied, integrating transcriptomics and metabolomics analysis, along with viability and cell cycle assays to study the molecular mechanisms that explain the anti-proliferative activity of this fruit by-product. After treatment for 48 and 72 h, the viability of HT-29 colon cancer cells was markedly affected, whereas minor effects were observed on normal human colon fibroblast cells. The bioactive extract was shown to arrest HT-29 cells in the S and G2/M phases of the cell cycle, which might be mediated by the inactivation of the FAT10 cancer signalling pathway among other genes identified as altered in the transcriptomic analysis. In addition, cellular redox homeostasis, as well as the polyamines pathway and methionine metabolism were found to be affected as suggested from the metabolomics data. Finally, the Foodomics integration enabled the identification of genes, such as MAD2L1, involved in the polyamine and glutathione metabolism, or the inactivation of the NUPR1 transcription factor, that might be related with the alteration of the intracellular ceramide levels in response to endoplasmic reticulum stress.

Asunto(s)

Antineoplásicos Fitogénicos/farmacología , Supervivencia Celular/efectos de los fármacos , Passiflora/química , Extractos Vegetales/farmacología , Semillas/química , Antineoplásicos Fitogénicos/química , Línea Celular , Proliferación Celular/efectos de los fármacos , Glutatión/metabolismo , Humanos , Metabolómica , Metionina/metabolismo , Extractos Vegetales/química , Poliaminas/metabolismo

RESUMEN

The aim of this study was to investigate the effect of the chronic administration of methionine (Met) and/or its metabolite, methionine sulfoxide (MetO), on the behavior and neurochemical parameters of young rats. Rats were treated with saline (control), Met (0.2-0.4 g/kg), MetO (0.05-0.1 g/kg), and/or a combination of Met + MetO, subcutaneously twice a day from postnatal day 6 (P6) to P28. The results showed that Met, MetO, and Met + MetO impaired short-term and spatial memories (P < 0.05), reduced rearing and grooming (P < 0.05), but did not alter locomotor activity (P > 0.05). Acetylcholinesterase activity was increased in the cerebral cortex, hippocampus, and striatum following Met and/or MetO (P < 0.05) treatment, while Na+, K+-ATPase activity was reduced in the hippocampus (P < 0.05). There was an increase in the level of thiobarbituric acid reactive substances (TBARS) in the cerebral cortex in Met-, MetO-, and Met + MetO-treated rats (P < 0.05). Met and/or MetO treatment reduced superoxide dismutase, catalase, and glutathione peroxidase activity, total thiol content, and nitrite levels, and increased reactive oxygen species and TBARS levels in the hippocampus and striatum (P < 0.05). Hippocampal brain-derived neurotrophic factor was reduced by MetO and Met + MetO compared with the control group. The number of NeuN-positive cells was decreased in the CA3 in Met + MetO group and in the dentate gyrus in the Met, MetO, and Met + MetO groups compared to control group (P < 0.05). Taken together, these findings further increase our understanding of changes in the brain in hypermethioninemia by elucidating behavioral alterations, biological mechanisms, and the vulnerability of brain function to high concentrations of Met and MetO.

Asunto(s)

Errores Innatos del Metabolismo de los Aminoácidos/complicaciones , Glicina N-Metiltransferasa/deficiencia , Hipocampo/patología , Trastornos de la Memoria/etiología , Trastornos de la Memoria/patología , Metionina/análogos & derivados , Especies Reactivas de Oxígeno/metabolismo , Acetilcolinesterasa/metabolismo , Errores Innatos del Metabolismo de los Aminoácidos/inducido químicamente , Errores Innatos del Metabolismo de los Aminoácidos/metabolismo , Animales , Catalasa/metabolismo , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Corteza Cerebral/patología , Femenino , Glutatión Peroxidasa/deficiencia , Glicina N-Metiltransferasa/metabolismo , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Masculino , Trastornos de la Memoria/metabolismo , Memoria a Corto Plazo/efectos de los fármacos , Metionina/metabolismo , Metionina/toxicidad , Ratas , Ratas Wistar , Memoria Espacial/efectos de los fármacos , Superóxido Dismutasa/deficiencia , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo