Allantoin Levels Research Articles

LC/MS/MS detection of uric acid oxidation metabolites to evaluate reactive oxygen and nitrogen species in vivo with application to pseudo-inflammation

Uric acid, a water-soluble antioxidant ubiquitous in human bodily fluids at relatively high concentrations, reacts with a variety of reactive oxygen and nitrogen species. Ours and other previous studies identified allantoin, oxaluric acid, triuret, and 5-N-carboxyimino-6-aminopyrimidine-2,4(3H)-dione as specific metabolites reactive against free radicals, singlet oxygen, peroxynitrite, and hypochlorous anion, respectively. We analyzed human plasma spiked with these products using high-performance liquid chromatography-tandem mass spectrometry. We observed recoveries of 40-110% and coefficients of variance within 7%. Samples remained stable at -80°C for at least 4 weeks, indicating the analytical method is sound. Detection of these metabolites in biological samples enables the identification of each species generated in vivo. We observed changes in the products in human blood during pseudo-inflammation induced by treatment with lipopolysaccharide. Levels of allantoin, oxaluric acid, triuret, and 5-N-carboxyimino-6-aminopyrimidine-2,4(3H)-dione increased after addition of lipopolysaccharide. The formation of singlet oxygen was confirmed by increased formation of Trp-derived cis-hydroxide ([2S,3aR,8aR]-3a-hydroxy-1,2,3,3a,8,8a-hexahydropyrrolo[2,3-b]indole-2-carboxylic acid) (cis-WOH). We believe our method will aid development of strategies to treat oxidative stress-associated diseases.

Disrupted NAD(P) Metabolism and Xanthine Dehydrogenase in a Stress-Induced Rat Model of Depression: NMR Metabolomics Insights.

Background: Clinical findings have shown a negative correlation between the severity of depressive symptoms and serum uric acid levels in men, yet the role of metabolic regulation in the pathophysiology of depression remains largely unknown. Methods: In this study, we utilized an acute restraint-stress-induced male rat model of depression to investigate biochemical changes through NMR-based metabolomics combined with serum biochemical analysis. Additionally, we employed qPCR, immunoblotting, and enzyme activity assays to assess the expression and activity of xanthine oxidoreductase, the rate-limiting enzyme in uric acid production. Results: Our findings indicate the following: (1) restraint stress is a valid method for inducing a depressive phenotype in rats; (2) depressive rats exhibit decreased NAD(P) levels in the liver and increased nicotinamide N-oxide and nicotinate levels in urine, accompanied by decreased levels of uric acid, allantoin, and allantoic acid in serum or tissues; (3) xanthine dehydrogenase activity is diminished in depressive rats without corresponding changes in gene or protein expression. Conclusion: The increased urinary excretion of NAD(P) precursors results in reduced hepatic NAD(P) levels, thereby suppressing NAD-dependent xanthine dehydrogenase activity and diminishing the production of uric acid and its downstream metabolites (allantoin and allantoic acid).

Salt stress alters root meristem definition, vascular differentiation and metabolome in Sorghum bicolor (L.) genotypes

Knowledge on salt tolerance requires further investigation, particularly in plants of agro-food interest. Sorghum is a potentially useful plant because it is a emerging food species that combines high levels of salt tolerance with interesting nutritional characteristics. In sorghum different genotypes respond differently to saline stress and the early events characterizing the salt stress tolerance are not yet fully understood. Moreover, the number of salt resistant genotypes needs to be extended. The genotypes Bianca and Tonkawa are two possible candidates for extending sorghum cultivation to soils characterized by high levels of salinity. The root is the first organ that responds to soil conditions, especially during the initial stages of plant developmental. The research aim was to analyse the root system responses to salt stress (NaCl) of Bianca and Tonkawa genotypes to identify the morpho-functional and metabolic changes that occur during the initial stages of the root system development and to use them as discriminating parameters for assessing the different plant’s susceptibility to the salt. The results showed that salt stress negatively affected many morphological and cyto-histological root parameters, from seed germination to root system establishment. The salt altered the root meristem organization and quiescent centre (QC) definition, but similarly in both genotypes. By contrast, it reduced primary root (PR) length and induced a more extended oxidative stress in the adventitious roots (ARs) and lateral root primordia (LRPs) of Tonkawa in comparison with Bianca. The stele area and the number of protoxylem and phloem elements in the ARs were more reduced in 150 mM NaCl-treated Tonkawa seedlings in comparison with those of Bianca. Moreover, the salt enhanced lignin deposition in protoxylem, early metaxylem and endodermis and changed the root metabolic profiles significantly increasing the levels of leucine, isoleucine, alanine, proline, trigonelline, allantoin and glutamine in Bianca compared to Tonkawa. Altogether, specific morpho-anatomical and metabolic differences between the genotypes were identified as discriminating markers of genotype salt susceptibility.

Liver Metabolomics Analysis Revealing Key Metabolites Associated with Different Stages of Nonalcoholic Fatty Liver Disease in Hamsters.

Nonalcoholic fatty liver disease (NAFLD) is not only the top cause of liver diseases but also a hepatic-correlated metabolic syndrome. This study performed untargeted metabolomics analysis of NAFLD hamsters to identify the key metabolites to discriminate different stages of NAFLD. Hamsters were fed a high-fat diet (HFD) to establish the NAFLD model with different stages (six weeks named as the NAFLD1 group and twelve weeks as the NAFLD2 group, respectively). Those liver samples were analyzed by untargeted metabolomics (UM) analysis to investigate metabolic changes and metabolites to discriminate different stages of NAFLD. The significant liver weight gain in NAFLD hamsters was observed, accompanied by significantly increased levels of serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Moreover, the levels of TG, LDL-C, ALT, and AST were significantly higher in the NAFLD2 group than in the NAFLD1 group. The UM analysis also revealed the metabolic changes; 27 differently expressed metabolites were detected between the NAFLD2 and NAFLD1 groups. More importantly, the levels of N-methylalanine, allantoin, glucose, and glutamylvaline were found to be significantly different between any two groups (control, NAFLD2 and NAFLD1). Receiver operating characteristic curve (ROC) curve results also showed that these four metabolites are able to distinguish control, NAFLD1 and NAFLD2 groups. This study indicated that the process of NAFLD in hamsters is accompanied by different metabolite changes, and these key differently expressed metabolites may be valuable diagnostic biomarkers and responses to therapeutic interventions.

Allantoin and Tissue Specific Redox Regulation in Mud Crab Scylla serrata under Varied Natural Water Physico-Chemical Parameters

Effects of varied sediment and physico-chemical properties of water on allantoin content in tissues of Scylla serrata and its role in antioxidant homeostasis were investigated. Tissues of crabs were sampled from different coastal natural habitats of S. serrata of India during summer, winter, and rainy seasons and were analyzed to detect the variation in allantoin content and lipid peroxidation (LPx) and oxidative stress (OS) neutralizing antioxidant factors. High allantoin content in hepatopancreas over muscle tissue was observed in all seasons and sampling sites. The correlation coefficient values between allantoin and environmental factors, i.e., temperature, pH, salinity, organic carbon, Mg, and Ca, strongly support the stress-induced allantoin variation level in tissues. The level of allantoin had a negative correlation with levels of ROS, which was probably due to the upregulation of the activity of major antioxidant enzymes and assisting enzymes such as glutathione peroxidase (GPx), glutathione reductase (GR), and biotransforming enzyme glutathione -s- transferases (GST). A significant seasonal variation in the level of allantoin was correlated with the activity of including superoxide dismutase, catalase, GPx, GR, and GST, which was clearly noticed as a function of abiotic factors. Additionally, the level of allantoin did not correlate with small antioxidant molecules, such as ascorbic acid and reduced glutathione. Discriminant function analysis revealed that the level of allantoin and CAT and GR activities were the major contributing factors for the clear discrimination of groups. Therefore, allantoin can be considered as a significant factor for the seasonal modulation of OS physiology in mud crab Scylla serrata.

Effects of feeding different dietary rates of mixed fodder beet tops-wheat straw silage on the performance of Holstein lactating cows.

Rations containing different rates of the mixed fodder beet tops-wheat straw silage (BS), instead of corn silage (CS), were given to 30 mid-lactation Holstein cows (all in parity 2) to measure the effects on feed consumption, milk production efficiency, milk chemistry, urinary purine derivatives (PD), blood chemistry, antioxidant levels, and in vitro methane (CH4) emission. The BS was prepared by mixing the fodder beet tops with wheat straw at a ratio of 9:1 based on fresh weight. The experimental design was completely randomized (one 28-d period with 21-d adaptation) using 30 cows (10 animals/treatment) and 3 treatments. The treatments were 1) a diet containing CS only (25g/100g DM) (CSD), 2) a diet containing 50% CS (12.5g/100g DM) and 50% BS (12.5g/100g DM) (CBSD), and 3) diet containing BS only (25g/100g DM) (BSD). Each animal (as an experimental unit) was housed individually in the tie stall and had ad libitum access to its diet. Dietary replacing 50% of CS with BS showed no significant differences in milk production, fat-corrected milk, fat and protein yields, feed efficiency, and apparent digestibility, however, these variables were less (P < 0.05) in the cows fed with BSD. Cows fed on BSD had less intakes of DM, organic matter, crude protein, and neutral detergent fiber but greater oxalic acid intake and blood urea-N, as compared to the other cows. Milk percentages of fat, protein, lactose, urea N, blood serum glucose, triglyceride, cholesterol, total protein, albumin, globulin, Ca, and P, as well as in vitro ruminal pH, were not affected by the diets. Saturated fatty acids concentration was less and monounsaturated FA and polyunsaturated FA (PUFA) was greater in the milk of cows receiving CBSD, compared to the other groups. The inclusion of both BS rates in the diet decreased the in vitro gas production, protozoa number, and CH4 emission in comparison to the control. Cows fed BSD had decreased levels of urinary allantoin, PD excreted or absorbed, and estimated microbial-N synthesis than the control and CBSD-fed groups. The milk and blood total antioxidant capacity (TAC) of the animals fed CBSD was the maximum among the cows. Overall, under the current experimental conditions, replacing 50% of dietary CS with BS did not affect milk production, but increased milk PUFA, as well as blood and milk TAC, and decreased in vitro CH4 emission, so it's feeding to lactating Holstein cows is recommended.

Serum metabolome differences associated with subclinical intramammary infection caused by Streptococcus agalactiae and Prototheca spp. in multiparous dairy cows

Mastitis is one of the most significant diseases in dairy cows and causes several economic losses. Somatic cell count (SCC) is often used as an indirect diagnostic tool for mastitis, especially for subclinical mastitis (SCM) where no symptoms or signs can be detected. Streptococcus agalactiae is one of the main causes of contagious mastitis, while Prototheca spp. is an alga inducing environmental mastitis that is not always correlated with increased milk SCC. The aim of this study was to evaluate the changes in the metabolomic profile of blood in relation to subclinical intramammary infection (sIMI) in dairy cows. In addition, differences due to the etiologic agent causing mastitis were also considered. Forty Holstein-Friesian dairy cows in mid and late lactation were enrolled in this study with a cross-sectional design. Based on the bacteriological examination of milk, the animals were divided into 3 groups: Group CTR (control group; n = 16); Group A (affected by SCM with IMI of Streptococcus agalactiae; n = 17); and Group P (affected by SCM with IMI of Prototheca spp.; n = 7). Blood samples were collected in tubes containing clot activator from jugular vein. The serum aliquot was stored until metabolomic analysis by 1H-NMR. Statistical analysis was conducted fitting a linear model with the group as fixed effect and SCC as covariate. Forty-two metabolites were identified and among them, 10 were significantly different among groups. Group A and P showed greater level of His and lactose, and lower level of acetate, Asn, and dimethylamine compared with Group CTR. Group A showed high level of Val, while the Group P showed also high level of Cit and methylguanidine, and lower level of 3-hydroxybutyrate, acetone, allantoin, carnitine, citrate, and ethanol. These metabolites were related to ruminal fermentations, energy metabolism, urea synthesis and metabolism, immune and inflammatory response, and mammary gland permeability. These results are suggestive of a systemic involvement of sIMI and that the metabolic profile of animals with SCM undergoes changes related to the etiological agent of mastitis.

Effect of Combined Infrared Hot Air Drying on Yam Slices: Drying Kinetics, Energy Consumption, Microstructure, and Nutrient Composition.

Using hot air drying (HAD) and combined infrared hot air drying (IR-HAD) test devices, the drying kinetics, unit energy consumption, color difference values, rehydration rate, microstructure, and changes in polysaccharide and allantoin contents of yam slices were examined at various temperatures (50 °C, 55 °C, 60 °C, 65 °C, and 70 °C). The findings demonstrated that each of the aforementioned parameters was significantly impacted by the drying temperature. IR-HAD dries quicker and takes less time to dry than HAD. The Deff of IR-HAD is higher than that of HAD at the same temperature and increases with the increase in temperature. The activation energy required for IR-HAD (26.35 kJ/mol) is lower than that required for HAD (32.53 kJ/mol). HAD uses more energy per unit than IR-HAD by a factor of greater than 1.3. Yam slices treated with IR-HAD had higher microscopic porosity, better rehydration, lower color difference values, and higher polysaccharide and allantoin levels than HAD-treated yam slices. The IR-HAD at 60 °C had the greatest comprehensive rating after a thorough analysis of the dried yam slices using the coefficient of variation method. Three statistical indicators were used to evaluate six thin-layer drying models, and the Weibull model was most applicable to describe the variation of drying characteristics of yam slices.

Hyperuricaemia Does Not Interfere with Aortopathy in a Murine Model of Marfan Syndrome.

Redox stress is involved in the aortic aneurysm pathogenesis in Marfan syndrome (MFS). We recently reported that allopurinol, a xanthine oxidoreductase inhibitor, blocked aortopathy in a MFS mouse model acting as an antioxidant without altering uric acid (UA) plasma levels. Hyperuricaemia is ambiguously associated with cardiovascular injuries as UA, having antioxidant or pro-oxidant properties depending on the concentration and accumulation site. We aimed to evaluate whether hyperuricaemia causes harm or relief in MFS aortopathy pathogenesis. Two-month-old male wild-type (WT) and MFS mice (Fbn1C1041G/+) were injected intraperitoneally for several weeks with potassium oxonate (PO), an inhibitor of uricase (an enzyme that catabolises UA to allantoin). Plasma UA and allantoin levels were measured via several techniques, aortic root diameter and cardiac parameters by ultrasonography, aortic wall structure by histopathology, and pNRF2 and 3-NT levels by immunofluorescence. PO induced a significant increase in UA in blood plasma both in WT and MFS mice, reaching a peak at three and four months of age but decaying at six months. Hyperuricaemic MFS mice showed no change in the characteristic aortic aneurysm progression or aortic wall disarray evidenced by large elastic laminae ruptures. There were no changes in cardiac parameters or the redox stress-induced nuclear translocation of pNRF2 in the aortic tunica media. Altogether, the results suggest that hyperuricaemia interferes neither with aortopathy nor cardiopathy in MFS mice.

Serum but not cerebrospinal fluid levels of allantoin are increased in de novo Parkinson’s disease

Oxidative stress supposedly plays a role in the pathogenesis of Parkinson’s disease (PD). Uric acid (UA), a powerful antioxidant, is lowered in PD while allantoin, the oxidation product of UA and known biomarker of oxidative stress, was not systematically studied in PD. We aim to compare serum and cerebrospinal fluid (CSF) levels of UA, allantoin, and allantoin/UA ratio in de novo PD patients and controls, and evaluate their associations with clinical severity and the degree of substantia nigra degeneration in PD. We measured serum and CSF levels of UA, allantoin, and allantoin/UA ratio in 86 PD patients (33 females, mean age 57.9 (SD 12.6) years; CSF levels were assessed in 51 patients) and in 40 controls (19 females, 56.7 (14.1) years). PD patients were examined using Movement Disorder Society-Unified Parkinson’s Disease Rating Scale (MDS-UPDRS), Montreal Cognitive Assessment (MoCA), Scales for Outcomes in Parkinson Disease-Autonomic (SCOPA-AUT), the University of Pennsylvania Smell Identification Test (UPSIT), one-night video-polysomnography, and dopamine transporter single-photon emission computed tomography (DAT-SPECT). Serum allantoin and allantoin/UA ratio were significantly increased in the PD group compared to controls (p < 0.001 and p = 0.002, respectively). Allantoin/UA ratios in serum and CSF were positively associated with the SCOPA-AUT score (p = 0.005 and 0.031, respectively) and RBD presence (p = 0.044 and 0.028, respectively). In conclusion, serum allantoin and allantoin/UA ratio are elevated in patients with de novo PD. Allantoin/UA ratio in serum and CSF is associated with autonomic dysfunction and RBD presence, indicating that higher systemic oxidative stress occurs in PD patients with more diffuse neurodegenerative changes.

Allantoin induces pruritus by activating MrgprD in chronic kidney disease.

Chronic kidney disease (CKD) is a disease with decreased, irreversible renal function. Pruritus is the most common skin symptom in patients with CKD, especially in end-stage renal disease. The underlying molecular and neural mechanism of CKD-associated pruritus (CKD-aP) remains obscure. Our data show that the level of allantoin increases in the serum of CKD-aP and CKD model mice. Allantoin could induce scratching behavior in mice and active DRG neurons. The calcium influx and action potential reduced significantly in DRG neurons of MrgprD KO or TRPV1 KO mice. U73122, an antagonist of phospholipase C, could also block calcium influx in DRG neurons induced by allantoin. Thus, our results concluded that allantoin plays an important role in CKD-aP, mediated by MrgprD and TrpV1, in CKD patients.

DETERMINATION OF ALLANTOIN LEVELS AS A RESULT OF OPTIMIZATION OF IONIC LIQUID-ULTRASOUND ASSISTED EXTRACTION OF COMFREY LEAVES BY TLC-DENSITOMETRY

Allantoin is a compound contained in comfrey leaves. This research aims to obtain optimal condition parameters of ionic liquid-ultrasound-assisted extraction (IL-UAE) comfrey leaves to attract allantoin compounds. Comfrey leaves are extracted with eight ionic liquids by ultrasound-assisted extraction (UAE) and screened for allantoin levels. The best ionic liquid from the screening results is optimized response surface methodology (RSM) with Box-Behnken design with three factors and three levels to determine the optimum extraction conditions for allantoin levels. Determination of levels is carried out with TLC densitometry. The results of the IL screening obtained [HMIM]Cl as the best IL in the evaluation of optimization of extraction parameters. The results of optimization of extraction condition parameters [HMIM]Cl obtained the highest allantoin levels on the sixth run of 306.396 μg / g of powder at the ratio of solvents to powders of 10 mL / g, concentrations [HMIM] Cl 1 mol / L and extraction time of 30 minutes.

Salivary metabolic signatures of carotid atherosclerosis in patients with type 2 diabetes hospitalized for treatment.

Atherosclerosis is a life-threatening disease associated with morbidity and mortality in patients with type 2 diabetes (T2D). This study aimed to characterize a salivary signature of atherosclerosis based on evaluation of carotid intima-media thickness (IMT) to develop a non-invasive predictive tool for diagnosis and disease follow-up. Metabolites in saliva and plasma samples collected at admission and after treatment from 25 T2D patients hospitalized for 2weeks to undergo medical treatment for diabetes were comprehensively profiled using metabolomic profiling with gas chromatography-mass spectrometry. Orthogonal partial least squares analysis, used to explore the relationships of IMT with clinical markers and plasma and salivary metabolites, showed that the top predictors for IMT included salivary allantoin and 1,5-anhydroglucitol (1,5-AG) at both the baseline examination at admission and after treatment. Furthermore, though treatment induced alterations in salivary levels of allantoin and 1,5-AG, it did not modify the association between IMT and these metabolites (p interaction > 0.05), and models with these metabolites combined yielded satisfactory diagnostic accuracy for the high IMT group even after treatment (area under curve = 0.819). Collectively, this salivary metabolite combination may be useful for non-invasive identification of T2D patients with a higher atherosclerotic burden in clinical settings.

Alterations in bone metabolites with age in C57BL/6 mice model.

Understanding the pathophysiology behind age-related diseases is an urgent need as the elderly population continues to grow. With age, there is a high risk of musculoskeletal deterioration and associated morbidity and mortality. Although the exact mechanism behind age-related degeneration is unknown, it is well established that alteration in cellular metabolism is one of the important contributing factors. Alteration in signaling pathways with age leads to the accumulation or depletion of several metabolites that play a vital role in musculoskeletal pathophysiology. This study aimed to identify age-related changes in bone tissue metabolites in C57BL/6 mice. We then correlated the differentially expressed metabolites with their functions in bone biology. In both aged males and females, hydroxyproline, glutamine, and alpha-linolenic acid levels were decreased. In aged females, Ornithine (p value = 0.001), L-Proline (p value = 0.008), Uridine (p value = 0.001), Aspartic Acid (p value = 0.004) levels were significantly decreased, and glutamate (p value = 0.002) was elevated. In aged males, N-acetyl-D-glucosamine (pvalue = 0.010), Adrenic acid (pvalue = 0.0099), Arachidonic acid (p value = 0.029) and Allantoin (p value = 0.004) levels were decreased. Metabolic pathway analysis revealed that purine and D-glutamine and D-glutamate metabolism were significantly altered in both sexes, while arginine biosynthesis in females and lipid metabolism in males were highly affected. These differences in metabolic signaling might be one of the reasons for the discrepancy in musculoskeletal disease manifestation between the two sexes. Understanding the role of these metabolites play in the aging bone will allow for new sex-specific targeted therapies against the progression of musculoskeletal diseases.

A Role for Allantoate Amidohydrolase (AtAAH) in the Germination of Arabidopsis thaliana Seeds

Seed dormancy is a very complex trait controlled by interactions between genetic and environmental factors. Nitrate is inversely correlated with seed dormancy in Arabidopsis. This is explained by the fact that seed dry storage (after-ripening) reduces the need for nitrogen for germination. When nitrate is absorbed by plants, it is first reduced to nitrite and then to ammonium for incorporation into amino acids, nucleic acids and chlorophyll. Previously, we showed that ALLANTOATE AMIDOHYDROLASE (AtAAH) transcripts are up-regulated in imbibed dormant seeds compared with after-ripened seeds. AAH is an enzyme in the uric acid catabolic pathway which catalyzes the hydrolysis of allantoate to yield CO2, NH3 and S-ureidoglycine. This pathway is the final stage of purine catabolism, and functions in plants and some bacteria to provide nitrogen, particularly when other nitrogen sources are depleted. Ataah mutant seeds are more dormant and accumulate high levels of allantoate, allantoin and urea, whereas energy-related metabolites and several amino acids are lower upon seed imbibition in comparison with Columbia-0. AtAAH expression could be detected during the early stages of seed development, with a transient increase around 8 d after pollination. AtAAH expression is the highest in mature pollen. The application of exogenous potassium nitrate can partly complement the higher dormancy phenotype of the Ataah mutant seeds, whereas other nitrogen sources cannot. Our results indicate that potassium nitrate does not specifically overcome the alleviated dormancy levels in Ataah mutant seeds, but promotes germination in general. Possible pathways by which AtAAH affects seed germination are discussed.

Quantification of allantoin and other metabolites of the purine degradation pathway in human plasma samples using a newly developed HILIC-LC-MS/MS method.

The development of a simple HILIC-LC-MS/MS method to quantify the plasma levels of allantoin, inosine, hypoxanthine, and adenosine, using stripped plasma for the bioanalytical method validation, was the purpose of this study. Chromatographic separation conducted using an XBridge BEH Amide column (2.1 × 150mm,3.5 μm) was achieved under gradient elution with two mobile phases: 0.1% formic acid-ACN (5:95) and 0.1% formic acid-ACN (50:50). Multiple reaction monitoring MS detection was performed using a triple quadrupole. The method validation experiments were performed according to the European Medicines Agency and the U.S. Food and Drug Administration guidelines. The lower LOQ was 50nM, 5nM, 20nM, and 2nM for allantoin, inosine, hypoxanthine, and adenosine, respectively. The recovery was repeatable and stable. The intraday precision ranged from 1.6% to 6.5%, while the interday precision ranged from 3.4% to 58.7%. Therefore, it is necessary to make a matrix-matched calibration curve each day to overcome this issue. Since the quality control samples' stability did not always comply with the guidelines, the samples need to be analyzed soon after collection.

Enterocyte synthesizes and secrets uric acid as antioxidant to protect against oxidative stress via the involvement of Nrf pathway

The gut is an important site to excreting uric acid (UA) in addition to the kidney. The gastrointestinal tract is constantly exposed to various potentially harmful substances, triggering intestinal oxidative damage. In the present study, the hypothesis that UA is can be synthesized to function as an antioxidant in the gut is evaluated. The synthesis and secretion of UA by enterocytes were analyzed in the presence of inosine, a precursor of UA, febuxostat (Fx), an inhibitor of xanthine oxidase (XOR), and H2O2. The regulation of Nrf2 pathway on UA secretion and transport were evaluated in the present of agonist (TBHQ) and inhibitor (ML385) of Nrf2. The in vivo result showed that UA and its oxidation product allantoin were presented in gut contents along the gastrointestinal tract and the highest level of UA and allantoin were detected in duodenum and jejunum respectively. The genes in the de novo purine nucleotide synthesis and salvage-catabolism pathways, and UA transporters were expressed in the intestinal tract. In the in vitro cultured enterocytes and everted gut sacs, inosine stimulated UA synthesis and secretion. H2O2 stimulated UA synthesis and secretion and meanwhile induced oxidative damage. UA attenuated H2O2-induced oxidative damage by Nrf2 pathway. UA secretion and transport were reduced by blocking Nrf2 with ML385, while increased by activating Nrf2 with TBHQ. This study provides new insights into the antioxidant effects if UA on intestinal lumen. The result suggests that activation of Nrf2 pathway is involved in the transportation and secretion of UA.

Patients with REM sleep behavior disorder have higher serum levels of allantoin

IntroductionRapid eye movement (REM) sleep behavior disorder (RBD) is associated with an increased risk of developing Parkinson's disease (PD). Low uric acid (UA) levels are associated with the risk of development and progression of PD. Allantoin is the major oxidation product of UA and is considered as a biomarker of oxidative stress. We aimed to compare serum levels of UA, allantoin, and allantoin/UA ratio in RBD patients with those in healthy controls, and to examine their associations with clinical severity. MethodsWe evaluated serum levels of UA, allantoin, and allantoin/UA ratio in 38 RBD patients (one female, mean age 66.8 (SD 6.3) years) and in 47 controls (four females, 66.8 (7.6) years). All RBD patients were assessed according to an examination protocol, which included structured interview, Montreal Cognitive Assessment (MoCA), Movement Disorder Society-Unified Parkinson's Disease Rating Scale (MDS-UPDRS), and dopamine transporter single-photon emission computed tomography (DAT-SPECT). The lower putaminal binding ratio from both hemispheres was used for analysis. ResultsMean serum allantoin concentration and allantoin/UA ratio were significantly increased in the RBD group compared to controls (2.6 (1.8) vs. 1.4 (0.7) μmol/l, p = 0.0004, and 0.008 (0.004) vs. 0.004 (0.002), p < 0.0001, respectively). There were no significant differences in UA levels between the two groups. No significant associations between any biochemical parameter and RBD duration, putaminal binding ratio on DAT-SPECT, MDS-UPDRS, or MoCA score were found. ConclusionSerum allantoin and allantoin/UA ratio are increased in RBD patients in comparison to controls, which may reflect increased systemic oxidative stress in prodromal synucleinopathy.

Monitoring the Transition Period in Dairy Cows through 1H NMR-Based Untargeted Metabolomics

The metabolic alterations associated with the increase in milk production make the transition period critical to the health of dairy cows, usually leading to a higher incidence of disease in periparturient animals. In this manuscript, we describe the use of NMR-based untargeted metabolomics to follow how these changes impact the serum metabolome in a group of 28 transition dairy cows with no initial clinical diseases. Principal component analysis (PCA) of serum 1H NMR data from four weeks before calving to 8 weeks after parturition allowed us to clearly identify four stages during the transition period. Pairwise comparisons using orthogonal partial least square discriminant analysis (OPLS-DA) and univariate data analysis led to the identification of 18 metabolites that varied significantly through these stages. Species such as acetate, betaine, and creatine are observed early after calving, while other markers of metabolic stress, including acetone, β-hydroxybutyrate (BHB), and choline, accumulate significantly at the height of milk production. Furthermore, marked variations in the levels of lactate, allantoin, alanine, and other amino acids reveal the activation of different gluconeogenic pathways following parturition. Concomitant with a return to homeostasis, a gradual normalization of the serum metabolome occurs 8 weeks after calving. Correlations of metabolite levels with dietary and metabolic adaptations based on animal parity could also be identified. Overall, these results show that NMR-based chemometric methods are ideally suited to monitor manifestations of metabolic diseases throughout the transition period and to assess the impact of nutritional management schemes on the metabolism of dairy cows.

A Systematic Review of the Effects of Skin-to-Skin Contact on Biomarkers of Stress in Preterm Infants and Parents.

Background:Premature infants and their parents experience significant stress related to separation and lifesaving procedures. While evidence suggests that skin-to-skin contact (SSC) is a stress-reducing intervention for both neonates and parents, the mechanisms that underlie its efficacy are not well understood.Objective:Purpose of this systematic review is to summarize the current state of knowledge on changes in biomarkers (ie, oxytocin [OT], cortisol, hypoxanthine, xanthine, uric acid, and allantoin), associated with SSC in premature infants and parents, that may reflect physiologic responses to stress.Methods:A comprehensive literature search was conducted from 1990 to 2020. Studies were selected using prespecified inclusion and exclusion criteria.Results:Of the 175 studies identified, only 19 are included in this review. Ten studies evaluated only infants, 2 evaluated only parents, and 7 evaluated for changes in biomarkers in both infants and parents. Cortisol was the most common biomarker evaluated. While changes in infants' cortisol levels were highly variable, in 55% of the parent studies, parent cortisol levels decreased following SSC. In both parents and infants, OT levels decreased following SSC. Only 1 study found that allantoin levels were significantly lower in infants who received SSC.Implications for Practice and Research:While evidence suggests the numerous benefits of SSC, additional research is needed to identify the optimal biomarker to determine the mechanisms that underlie these effects. The use of novel biomarkers (eg, gene expression changes microbiome) may provide new insights into the mechanisms that underlie the efficacy of SSC.Video available at: https://journals.lww.com/advancesinneonatalcare/Pages/videogallery.aspx?autoPlay=false&videoId=48