Allantoin Content Research Articles

The effect of presowing saturation with molybdatenum and presence of nitrate on the allantoin content in sprouted wheat grain

The effect of presowing saturation with molybdatenum and presence of nitrate on the allantoin content in sprouted wheat grain

Influence of Nitrogen Deficiency on Ureide Mediated Stress Resistance in Arabidopsis thaliana

The relationship between nitrogen availability and presence of heavy metal stress appear to affect development and growth of Arabidopsis thaliana plants. In this work, Arabidopsis thaliana seedlings were exposed to media containing varying concentrations of nitrogen in form of NH4NO3 or KNO3 and cadmium in the form of CdCl2. When exposed to increasing cadmium stress, aln mutant lines, lacking a functional allantoinase enzyme, displayed enhanced growth and development when compared to wild type, most likely due to their high allantoin content. Increasing nitrogen concentrations appeared to decrease the amount of senescing tissue caused by exposure to cadmium stress. Growth appeared to be slightly increased in both wild type and aln mutant lines grown in media containing KNO3 as the nitrogen source, thereby supporting the importance of nitrogen source on plant growth.

블랜칭 처리에 의한 신선편이 둥근마의 저장 중 품질 특성

This study was carried out to investigate shelf-life and quality of fresh-cut Dungkunma (Dioscorea bulbifera) in order to elevate utilization of Dungkunma a fresh food. Before vacuum-packaging (in polyethylene/polypropylene film (100μm, 15×20 cm, 75±2 cmHg) and storaging at 2℃, Dungkunma was peeled out and cut to dice type (2.0±0.5 cm3), and then washed and blanched using hot water (at 90±2℃ with 2% NaCl solution for 30 sec). Blanched Dungkunma was pre-dried at room temperature, 40℃ and 50℃ for removing surface water. Each peeled dice Dungkunma was packed 50 g in polyethylene/polypropylene film (100 μm, 15×20 cm) with vacuum treatment (75±2 cmHg) and stored at 2℃ for 90 days. Hardness and adhesiveness of Dungkunma blanched by 2% NaCl and pre-dried at 50℃ (SB50) were the highest, but changes were the least during storage. Lightness and yellowness of stored Dungkunma in all treatments decreased slightly while redness increased during storage. Changes of color of SB50 was the least. Total concentration of aerobic bacteria in SB50 was 1.88±0.18 log CFU/g during 90 days and E. coli was detected in all treatments during whole storage periods. Dioscin and allantoin contents of SB50 were virtually unchanged during the storage. Consequently, the results of this study suggest that vacuum packaged Dungkunma after blanching using 2% NaCl solution could be effective to prolong the quality of fresh-cut Dungkunma.

Allantoin accumulation mediated by allantoinase downregulation and transport by Ureide Permease 5 confers salt stress tolerance to Arabidopsis plants.

Allantoin, a metabolite generated in the purine degradation pathway, was primarily considered an intermediate for recycling of the abundant nitrogen assimilated in plant purines. More specifically, tropical legumes utilize allantoin and allantoic acid as major nodule-to-shoot nitrogen transport compounds. In other species, an increase in allantoin content was observed under different stress conditions, but the underlying molecular mechanisms remain poorly understood. In this work, Arabidopsis thaliana was used as a model system to investigate the effects of salt stress on allantoin metabolism and to know whether its accumulation results in plant protection. Plant seedlings treated with NaCl at different concentrations showed higher allantoin and lower allantoic acid contents. Treatments with NaCl favored the expression of genes involved in allantoin synthesis, but strongly repressed the unique gene encoding allantoinase (AtALN). Due to the potential regulatory role of this gene for allantoin accumulation, AtALN promoter activity was studied using a reporter system. GUS mediated coloration was found in specific plant tissues and was diminished with increasing salt concentrations. Phenotypic analysis of knockout, knockdown and stress-inducible mutants for AtALN revealed that allantoin accumulation is essential for salt stress tolerance. In addition, the possible role of allantoin transport was investigated. The Ureide Permease 5 (UPS5) is expressed in the cortex and endodermis of roots and its transcription is enhanced by salt treatment. Ups5 knockout plants under salt stress presented a susceptible phenotype and altered allantoin root-to-shoot content ratios. Possible roles of allantoin as a protectant compound in oxidative events or signaling are discussed.

Urinary Allantoin Is Elevated in Severe Intraventricular Hemorrhage in the Preterm Newborn.

Germinal matrix intraventricular hemorrhage (IVH) is the most common type of intracranial hemorrhage observed in preterm neonates. It is a precursor of poor neurocognitive development, cerebral palsy, and death. The pathophysiology is not well defined, but damage to the fragile germinal matrix vasculature may be due to free radicals generated during inflammation and as a consequence of ischemia followed by reperfusion. Assessment of the oxidative stress status in these infants is therefore important. Urinary allantoin concentration was measured in preterm neonates as a marker of oxidative stress associated with IVH. Urine was collected from 44 preterm neonates at four time points between 24 and 72 hours of life (HOL), and the allantoin content was determined by gas chromatography mass spectrometry (GCMS). Records were retrospectively reviewed, and the incidence and severity of IVH was categorized as follows: no IVH (n = 24), mild (grade 1-2) IVH (n = 13), and severe (grade 3-4) IVH (n = 7). Neonates with severe IVH showed significantly elevated allantoin levels vs subjects with no IVH from 36 HOL (0.098 ± 0.013 μmol and 0.043 ± 0.007 μmol, respectively, p = 0.002). The allantoin concentration remained elevated even at 72 HOL (0.079 ± 0.014 μmol and 0.033 ± 0.008 μmol, respectively, p = 0.021). There were no significant differences in allantoin levels in the no IVH and mild IVH groups. IVH was diagnosed by head imaging on average at about 11th postnatal day. Urinary allantoin levels were significantly elevated during the first 3 days of life in the neonates subsequently diagnosed with severe IVH, suggesting that oxidative stress might be a crucial factor in IVH pathogenesis. Further studies are needed to assess the usefulness of urinary allantoin in early identification of preterm infants at risk for or with severe IVH and monitoring of the response to interventions designed to prevent or treat it.

The identification and quantification of bioactive compounds from the aqueous extract of comfrey root by UHPLC-DAD-HESI-MS method and its microbial activity

In this study the qualitative, quantitative and microbial determination of the aqueous extract of comfrey root was done. The qualitative and quantitative analyses were done by the UHPLC-DAD-HESI-MS method. As major bioactive compounds, allantoin, rosmarinic acid and ellagic acid were identified and their quantification was also done. The obtained results showed a high content of allantoin, ellagic acid and rosmarinic acid (8.91, 7.4 and 12.8%, respectively) which indicated that the comfrey root can be used as a source for the isolation of these three compounds. The results obtained by the determination of the antimicrobial activity showed that Escherichia coli ATCC8739 and Salmonela typhimirium ATCC6538 were most sensitive to the aqueous extract of comfrey root. The results showed that allantoin did not express the antimicrobial activity on all the investigated bacteria species, and based on this it can be concluded that allantoin is not responsible for the antimicrobial activity of the aqueous extract of comfrey root.

Changes in main nutrients and medicinal composition of Chinese yam (Dioscorea opposita) tubers during storage.

The changes in the main nutrient and medicinal components during the storage of the Chinese yam (Dioscorea opposita) tubers were studied. The harvested tubers were stored under ambient conditions (10°C to 18°C, 60% to 80% Relative Humidity) and cold temperature and packaged conditions (4°C, 60% to 65% Relative Humidity) for 45day. The allantoin, starch, total alcohol-soluble sugar, reducing sugar, protein, and moisture contents of the samples were evaluated. Their amylase activities were also investigated. Results of ambient conditions indicated that, during storage, moisture decreased by 67.96% to 56.51%, and total sugars, reducing sugars, and protein increased by 6.49% to 9.81%, 1.7% to 2.27%, and 13.02% to 14.55%, respectively. Starch and enzyme activities increased during the early days of storage and progressively decreased, and the content of allantoin changed in volatility. The changes were more significant at cold temperatures and packaged conditions than at ambient conditions. This result suggests that after-ripening occurred in the early stages of Chinese yam tubers, which positively affected the nutritional potential of the tubers by a marked increase in nutrients. Low-temperature sweetening greatly affects the nutritional potential of tubers by a series of complicated interactions between starch and sugars at 4°C.

Distribution and Function of Allantoin (5-Ureidohydantoin) in Rice Grains

Despite increasing knowledge of allantoin as a phytochemical involved in rice, relatively little is known about its distribution and function in rice grains. In this study, allantoin was quantified in 15 Chinese rice grains, and its contents varied with grain fraction, cultivar, and genotype. Bran always had the highest allantoin level, followed by brown rice and milled rice. Hull contained the lowest allantoin content. Allantoin in japonica bran ranged from 70 to 171 μg/g but rarely exceeded 100 μg/g in indica bran. There was a positive relationship between allantoin level in grains and seedling survival in seedbeds under low temperature or water deficit. Exogenous allantoin stimulated plant growth, increased soluble sugar and free proline contents, and decreased malondialdehyde content in rice seedlings. However, allantoin did not show any antioxidant activity through free radical-scavenging capacity, reducing power, linoleic acid peroxidation inhibition, and chelating activity. The results suggest that allantoin in rice grains may play some roles in providing plant stress protection but not serving as a beneficial health antioxidant.

Lactobacillus acidophilus fermented yam (Dioscorea opposita Thunb.) and its preventive effects on gastric lesion

This study was carried out to find the possibility of fermenting yam (Dioscorea opposita Thunb.) with Lactobacillus acidophilus and the protective role on stomach lesion of rats. The solutions of raw and extracted yam used were 2 and 6%, and they were fermented by L. acidophilus at 37°C for 0, 12, 24, and 36 h. At intervals (0, 12, 24, and 36 h), viscosity, titratable acidity (TA), allantoin, and diosgenin were examined. In the results, it was observed the most desirable data at 12 h with 6% fermented raw yam (FRY) without any change in the content of allantoin and diosgenin. In the animal study by rats (Sprague-Dawley), the inhibition ratios on gastric lesion were 53.41% in FRY. Based on these data, it confirmed that raw yam powder fermented by L. acidophilus would be a functional food with the supplement of viable cells and nutraceutical component of allantoin and diosgenin.

Effect of Dioscorea opposita Thunb. (yam) supplementation on physicochemical and sensory characteristics of yogurt

A study was conducted to examine the physicochemical, microbial, and sensory properties of yogurt made by supplementing powdered yam Dioscorea opposita Thunb. (YPT) at different concentrations (0.2, 0.4, 0.6, and 0.8%, wt/vol) into milk, which was pasteurized and then fermented at 43°C for 6h and stored for 16 d. The pH values of all samples decreased, whereas viscosity values and mean microbial counts increased during storage. The L* and a* color values (indicators of lightness and redness, respectively) of yogurt samples were not remarkably influenced by adding YPT, whereas the b* values (indicating yellowness) significantly increased with the addition of YPT at all concentrations at 0 d of storage, probably due to the original yellow color of yam powder. In functional component analyses, when the concentration of YPT increased, the amount of allantoin and diosgenin proportionally increased. The content of allantoin was 3.22 and diosgenin 4.69 μg/mL when 0.2% (wt/vol) YPT was supplemented and did not change quantitatively during the storage period (16 d). The sensory test revealed that the overall acceptability scores of YPT-supplemented yogurt samples (0.2 to 0.6%, wt/vol) were quite similar to those of the control throughout the storage period of 16 d. Based on the data obtained from the present study, it was concluded that the concentrations (0.2 to 0.6%, wt/vol) of YPT could be used to produce YPT-supplemented yogurt without significant adverse effects on physicochemical, microbial, and sensory properties, and enhance functional components from the supplementation.

Physicochemical Properties of Dioscorea opposita Thunb. Solution Fermented by Lactobacillus bulgaricus

The present study was designed to determine the physicochemical properties of yam (Dioscorea) solution fermented by L. bulgaricus, the protective effect of fermented Dioscorea opposita Thunb. on stomach lesions, and the possibility of its application to fermented dairy products. When the concentration of yam increased, the pH increased in raw powder samples but not in extracted powder samples. In 6% raw powder containing the sample, the viable cell counts showed a dramatic increase, whereas those in extracted yam powder significantly decreased in all concentrations during 32 h of fermentation. The viscosity of extracted yam powder showed a lower value than that of raw powder, which was maintained throughout the fermentation. The solution of Dioscorea opposita Thunb. raw powder showed a significantly higher allantoin content compared with the extracted powder (p<0.05). At 2% of added Dioscorea opposita Thunb., 0.65 mg/mL was contained in both samples; however, when 6% of Dioscorea opposita Thunb. was added, the amount of diosgenin increased up to 1.9 mg/mL. When 200 mg of powder/kg body wt. was injected, gastric lesions were apparently reduced compared with those of control rats. With 100 mg/kg body wt. injected into rats, the protective effect was a little lower than that in 200 mg/kg body wt. Based on these data, the present study indicated that yam powder fermented by L. bulgaricus showed a protective effect on gastric lesions in rats; therefore, it may be included as an added ingredient in yogurt manufacture.

Caffeine Biosynthesis and Degradation in Tea [Camellia sinensis (L.) O. Kuntze] is under Developmental and Seasonal Regulation

To study caffeine biosynthesis and degradation, here we monitored caffeine synthase gene expression and caffeine and allantoin content in various tissues of four Camellia sinensis (L.) O. Kuntze cultivars during non-dormant (ND) and dormant (D) growth phases. Caffeine synthase expression as well as caffeine content was found to be higher in commercially utilized tissues like apical bud, 1st leaf, 2nd leaf, young stem, and was lower in old leaf during ND compared to D growth phase. Among fruit parts, fruit coats have higher caffeine synthase expression, caffeine content, and allantoin content. On contrary, allantoin content was found lower in the commercially utilized tissues and higher in old leaf. Results suggested that caffeine synthesis and degradation in tea appears to be under developmental and seasonal regulation.

Changes in Allantoin and Arginine Contents in Dioscorea opposita 'Tsukuneimo' during the Growth

The ureides, allantoin and allantoic acid, are believed to play an important role in the storage and translocation of nitrogen in higher plants. Arginine is a major component in the free amino acid pool in some plants. To investigate the behavior of ureides and free amino acids, especially arginine, in developing Dioscorea opposita plants 'Tsukuneimo', their contents as well as those of water and nitrogen in leaves, stems and tubers were quantitatively analyzed. The content of allantoin in the leaves decreased after any fertilization, while that in the stems did not increase after the 1st fertilization, 96 days after planting; it significantly increased after the 2nd fertilization, 26 days later. Allantoin content in tubers remained constant during tuber development. Arginine content in leaves and stems during the growth of 'Tsukuneimo' were lower than 0.15 and 4.8 μmol·g-1FW, respectively. The time course of arginine in stems content was similar to that of allantoin. In tubers, arginine content at 170 days after planting increased to reach 14 times that at 142 days but then decreased, demonstrating that D. opposita transiently accumulates arginine in stems as well as allantoin in leaves and stems but only accumulates the latter in developing tubers.

Change in the allantoin content of Dioscorea (tsukuneimo) during cooking

Change in the allantoin content of Dioscorea (tsukuneimo) during cooking

Quantitative Analysis of Allantoin in Fresh Tubers of Dioscorea opposita 'Tsukuneimo'

Allantoin in fresh tuber of Dioscorea opposita 'Tsukuneimo' was quantitatively analyzed because the compound is thought to prevent inflammation and ulcers in humans as well as to play an important role for storage and translocation of nitrogen in higher plants. The content of ureide (allantoin and allantoic acid) in 'Tsukuneimo' was determined by the modified method of spectrophotometry with phenylhydrazine, whereas the content of allantoin was determined by HPLC. Analyses revealed that 'Tsukuneimo' tubers contained 2.6mg·gFW-1 of allantoin, but leaves or stems had undetectable levels. D. opposita, 'Yamatoimo' and 'Nagaimo', and D. japonica 'Jinenjo' contained 2.3, 0.47 and 1.2 mg·gFW-1 of allantoin in the tuber, respectively, but negligible amount of allantoic acid in the tuber. Furthermore, Ipomoea batatas and Solanum tuberosum contained less than 0.1 mg·gFW-1 of allantoin and a negligible amount of allantoic acid. Colocasia antiquorum, 'Ishikawawase' and 'Kyouimo' had no detectable amounts of allantoin or allantoic acid.

Influence of photoperiod on the accumulation of allantoin in comfrey plants

In order to evaluate the influence of photoperiod on the accumulation of allantoin in comfrey (Symphytum officinale L.), a well known medicinal plant, an experiment was conducted during July and August, 1998. Cuttings obtained from 90 day old plants were submitted to four photoperiods (8, 12, 16 and 20 hours). After 60 days, allantoin content in roots and rhizomes was evaluated. The results showed that increases in photoperiod promoted an increment in the average content of allantoin in roots (0.06%, 0.303%, 1.213% and 4.78%). On the other hand, in rhizomes, allantoin accumulation decreased (9.65%, 7.14%, 0.55%) when the photoperiod was increased from 8 to 12 and 16 hours, respectively, stabilizing on a 20-hour photoperiod (0.53%). Plants cultivated under field conditions presented 2.55% and 2.63% allantoin content in rhizomes and roots, respectively. Based on the fact that in comfrey the roots are considered to be sites of allantoin synthesis, the results demonstrated that photoperiod could influence both the synthesis of allantoin in these organs as well as its accumulation in the rhizomes.

Quantitative separation of uric acid and allantoin from rat liver tissue

A simple procedure is described for the assay of liver uric acid and allantoin and their specific radioactivity after administration of a radioactive precursor. Uric acid was quantified by the uricase reaction in liver trichloroacetic acid (TCA) extracts. The ‘true’ allantoin content of the liver could be estimated only after precipitation with Hg-acetate, a step by which the standard allantoin was also quantitatively recovered. Crude extracts lead to the evaluation of ‘apparent’ allantoin. For the determination of specific radioactivity, the Hg-acetate precipitate was further purified by ion-exchange chromatography. The purity of the two metabolites was confirmed by ultraviolet absorbance spectra. HPLC, constancy of specific radioactivity and the absence of amino acids. The incorporation of [ 14c]formate into uric acid and allantoin in the liver was studied by this procedure. The radioactivity in allantoin was several-fold higher than that in uric acid up to 60 min after administration of the precursor. This quite unexpected result is not easily explained on the basis of current knowledge.

Pulmonary dysfunction in gold miners with reactive airways.

In a cross sectional study of a working population of black South African gold miners a sample of 1197 older miners was examined. Airway reactivity determined by a greater than 10% response of the forced expiratory volume in one second (FEV1) to inhaled salbutamol was detected in 139 (12%) of the men. No relation was found between the extent of exposure to the underground environment and the frequency of airway reactivity. There was evidence of increased chronic airflow limitation in the men with reactive airways. This increase persisted after controlling for age, tobacco smoking, and for the presence and degree of silicosis. On average, the presence of reactive airways was associated with reduced (after bronchodilator) forced vital capacity (FVC) of 118 ml, 95% confidence interval (CI) 16 to 220 ml, FEV1 of 168 ml (95% CI 74-262 ml), FEV1/FVC% of 3% (95% CI 2.3-3.7%), and maximal mid-expiratory flow of 300 ml/s (95% CI 86-514 ml/s). Although there was no evidence that airway reactivity was induced by this occupational exposure, those with reactive airways appeared to be more susceptible to the adverse effects of the underground environment on the bronchial tree.

Response of Leucaena/Rhizobium symbiosis to mineral nutrients in Southwestern Nigeria

Pot and field experiments carried out at the International Institute of Tropical Agriculture (IITA) and at Fashola, Southwestern Nigeria, examined the effect of inoculation and N, P and micronutrients on nodulation and growth ofLeucaena leucocephala (Lam.) de Wit. In pot studies all parameters measured, except the percentage of nitrogen in shoots, were improved by inoculation, nitrogen and phosphorus. Micronutrients increased only nitrogen and allantoin contents. Interactions between inoculation and P, N and micronutrients on nodulation and growth of leucaena were observed. The effect of inoculation and fertilization with phosphorus or micronutrients was further investigated in field experiments. Establishment of uninoculated and unfertilized leucaena was poor at both locations due to low soil fertility and the presence of only a few native leucaena rhizobia. At one site, only inoculated plants were nodulated, while at the other, all plants produced nodules. Shoot dry weight, total nitrogen and phosphorus of inoculated plants were statistically equal to nitrogen-fertilized plants. Uninoculated plants were stunted. Generally, micronutrients did not influence nodulation, total nitrogen or growth of leucaena. They had only a positive effect on nitrogenase activity. Phosphorus increased total nitrogen and phosphorus uptake and plant growth. A 75% increase in shoot dry weight was obtained when 80 kg P ha−1 was applied to inoculated leucaena with Rhizobium strain IRc 1045. Inoculated plants contained more allantoins than uninoculated ones but no significant correlation was found between these compounds and other parameters of N fixation.

3 DETERMINATION OF ALLANTOIN IN MAMMALIAN TISSUES

Allantoin - the key compound in purine catabolism - is determined in biological systems using the Rimini's reaction, which involves treatment with 0.1 N alkali (100°C) and formation of allantoic acid, 0.025 N HCl (100°C), and production of urea and glyoxylic acid; final determination of the latter compound as a dinitrophenylhydrazine derivative. The reaction is subject to several interferences. The purification of allantoin is also difficult and thus it cannot be analyzed after labelled precursor administration. We evaluated the allantoin content in rat tissue extracts using two procedures: (1) allantoinase treatment (instead of alkali), subsequently proceeding as described above, (2) NaOH and HCl treatment, but a final determination of urea with urease and glutamate dehydrogenase. Results were lower compared with the conventional dinitrophenylhydrazine reaction. Precipitation of allantoin with Hg-acetate (followed by ion -exchange chromatography) guarantees excellent recovery of pure allantoin from tissue extracts. This has proved to be a simple procedure for determination of true allantoin content and of its labelling with isotopes for kinetic studies in different tissues.