Trp Levels Research Articles

Effects of Turmeric Rhizome Powder Supplemented Diet on Indomethacin-induced Toxicity in Wistar Rats

Turmeric (Curcuma longa) L rhizome powder (TRP) was commonly reported to have antiulcerogenic and non-toxicity effects. However, the scientific evidence showing the effectiveness of turmeric in the treatment of gastric ulcer and its non-toxicity effect are controversial. The aim of the study was to investigate the effects of different percentages of turmeric rhizome powder supplemented diet on toxicity induced by indomethacin in Wistar rats. This study investigated the effects of TRP formulated diet on the activities of blood enzymes in indomethacin-induced gastric ulcerated Wistar rats. This investigation was carried out through a 28-day experiment using corn-starch flour meal-based diet containing four levels of TRP (1%, 2%, 5% and 10%) as treatments with five replicates in a completely randomized design. The remaining three groups were fed with basal diet, one group received standard drug, another received no treatment but induced while the last group received no treatment and not induced. 35 male Wistar rats weighing 150-200 g were housed in seven cages and received feed and water ad-libitum. At the end of the experiment, all the animals were sacrificed, blood and some organs were collected and evaluated for hepatoxicity and nephrotoxicity induced by indomethacin (60mg/kg bw). Measurements of serum, kidney and liver alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were performed. Statistical evaluation of the results at a p < 0.05 showed significantly elevated values (P ≤ 0.05) of these enzymes in the kidney of rats in groups B, C, E and G when compared with group F. Increase in kidney ALT and AST activities of animals in groups A, C and D was detected in comparison with group F. Furthermore, there was increase in hepatic ALT and ALP activities of animals in groups A, B, C, D and E compared with animals in group F but a significant increase (P ≤ 0.05) in group G in comparison with group F. Likewise, there was significant increase (P ≤ 0.05) in liver AST activities of rats in groups A, B, C, E and G compared with group F, however, insignificant increase (P ≤ 0.05) was observed in animals in group D in comparison with rats in group F. Indomethacin induced rats (group F) showed a significant increase in serum levels of ALT, AST and ALP compared with rats in groups E and G in all the groups but those fed on 1%, 2%, 5% and 10% turmeric supplemented diet showed decrease in comparison with group F. The increased levels of these enzymes in the serum of animals in ulcerogenic group and some in group D (10%TRP group) could be a sign of tissue injury due to relative toxicity of indomethacin induction in animal model and deleterious effect of turmeric rhizome powder at large concentration. However, decrease levels of these enzymes in pretreated groups could indicate the attenuating potential of turmeric at moderate dose against toxicity effect of indomethacin induction. Therefore, turmeric rhizome powder should be consumed with caution and its percentage in the whole recipe should not be up to 10% (100g/kg).

Does the type of exercise affect tryptophan catabolism in horses?

Tryptophan (Trp) is an essential amino acid which metabolises via the kynurenine pathway to generate a number of bioactive substances referred to as kynurenines. Among those are 3-hydroxykynurenine (3-HKyn) and quinolinic acid, which are neurotoxic, as well as kynurenic acid (Kyna) and xanthurenic acid (XA), which, similarly to nicotinamide (NAm), show neuroprotective and anti-depressive effects. Routine exercise is known to modulate Trp metabolism in skeletal muscle and is thus believed to reduce the risk of depressive states in humans and laboratory animals. Analogously, it was hypothesised that exercise can influence Trp metabolism in horses as well. The aim of this study was to evaluate the influence of two different types of exercise on Trp metabolism in horses of the same breed. A total of 32 purebred Arabian horses were involved in the study. The 22 three-year-old racehorses were subjected to short-time intense exercise. Ten other horses were made to perform endurance competitions at a distance of 80 km. Blood samples were collected at rest and following the end of the exercise period. Plasma concentrations of Trp, kynurenine (Kyn), Kyna, 3-HKyn, XA and NAm were determined using Ultra-High Performance Liquid Chromatography-Electrospray Ionisation-Tandem Mass Spectrometry. Short-time intense exercise led to an increase in plasma concentrations of Kyn, Kyna and XA. The endurance effort induced an increase in Kyna and a decrease in Trp and NAm levels. Both types of exercise, short-time intensive exercise and endurance exercise induced an increase in Trp metabolites, especially Kyna, and did not induce an increase in Trp level. Thus, from a pathophysiological perspective of the kynurenine pathway’s influence on mental state, both types of exercise induced beneficial effects in horses.

A comprehensive analysis of TDO2 expression in immune cells and characterization of immune cell phenotype in TDO2 knockout mice.

Tryptophan 2,3-dioxygenase (TDO2) was an initial rate-limiting enzyme of the kynurenine (Kyn) pathway in tryptophan (Trp) metabolism. We undertook this study to determine a comprehensive analysis of TDO2 expression in immune cells and assess the characterization of immune cell phenotype in TDO2 knockout mice. The expression of TDO2 in various tissues of DBA/1 mice was detected by quantitative real-time PCR (qPCR) and immunohistochemistry. Both flow cytometry and immunofluorescence were used to analyze the expression of TDO2 in immune cells. Furthermore, TDO2 knockout (KO) mice were generated by CRISPR/Cas9 technology to detect immune cell phenotype. TDO2 protein level in liver was tested by western blot. High-performance liquid chromatography was used to detect the level of Trp and Kyn. Flow cytometry was used to test the proportions of splenic lymphocyte subsets in wild-type (WT) and TDO2 KO mice. We found that TDO2 was expressed in various tissues and immune cells, and TDO2 staining was mainly observed in the cytoplasm of cells. There was no difference in the development of immune cells between TDO2 KO mice and WT mice, including T cells, B cells, memory B cells, plasma cells, dendritic cells, and natural killer cells. Interestingly, the reduced M1/M2 ratio was observed in the peritoneal macrophages of TDO2 KO mice. Taken together, these findings enriched the known expression profile of TDO2, especially its expression in immune cells. Our study suggested that TDO2-mediated Trp-Kyn metabolism pathway might be involved in the immune response.

High levels of Tryptophan reduce cell wall or membrane stress tolerance in Saccharomyces cerevisiae.

Tryptophan (Trp) is a proteinogenic aromatic amino acid; however, high levels of Trp are toxic in animals and yeast with unknown mechanisms. Previously, we suggested that aromatic aminotransferase Aro9 is important for excess Trp degradation. Besides, Schroeder and Ikui showed that aro9Δ is sensitive to membrane stress by sodium dodecyl sulfate. Therefore, Trp accumulation may reduce the cell wall or membrane (CW/M) stress tolerance through participation of cell wall integrity (CWI) pathway, which detects and responds to CW/M perturbations. In this study, we found that yeast mutants of the CWI mitogen-activated protein kinase cascade were susceptible to excess Trp. Also, the Trp degradation deficient mutant aro8Δ aro9Δ cells, in which Trp accumulation was confirmed, were sensitive to several CW/M stresses. These results indicated that accumulation of Trp is adverse for the CW/M stress resistance and may disturb appropriate signal transduction responding to the stress.

Dietary tryptophan supplementation does not affect growth but increases brain serotonin level and modulates the expression of some liver genes in zebrafish (Danio rerio).

This study aimed at assessing the effects of the dietary tryptophan (Trp) supplementation on growth and feed utilization, brain serotonin content, and expression of selected liver genes (involved in the liver serotonin pathway, protein synthesis degradation, and antioxidant activity) in zebrafish. A growth trial was conducted with zebrafish juveniles fed five experimental isoproteic (40%DM) and isolipidic (8%DM) fishmeal-based diets containing graded levels of Trp: a Trp-non-supplemented diet (diet Trp0, with 0.22% Trp) and four Trp-supplemented diets containing 2-16 times higher Trp content (diets Trp2, Trp4, Trp8, and Trp16 with 0.40, 0.91, 2.02, and 3.34% Trp, respectively). Diets were tested in quadruplicate, with fish being fed twice a day, 6days a week for 6weeks to apparent visual satiation. At the end of the trial, growth performance and feed utilization were assessed, and fish from all experimental groups were sampled for whole-body composition analysis. In addition, fish fed low (Trp0), medium (Trp4), and high (Trp16) Trp diets were also sampled for analysis of brain serotonin content and liver gene expression. Tested tryptophan levels did not influence growth performance nor feed intake. However, values of energy and nitrogen retention as well as body energy content indicate a better feed utilization with diets containing around 0.9% and 2.0% DM Trp. Brain serotonin content increased with increasing dietary tryptophan levels. In addition, regarding liver genes, dietary treatment had a modulatory effect on the expression of Htr1aa and Htr2cl1 genes (encoding for serotonin receptors), TPH1a gene (encoding for tryptophan hydroxylase, the rate-limiting enzyme in the synthesis of serotonin from tryptophan), TOR gene (involved in protein synthesis), and Keap1 gene (involved in antioxidant responses).

Identification of biomarkers on kidney failure by Raman spectroscopy

AbstractChronic kidney disease (CKD) is a global health problem. Our study employed Raman spectroscopy for the first time to analyze potential biomarker such as tryptophan (Trp) on kidney failure and show the difference between normal and CKD groups. Adenine model of CKD characterized most of the physiological changes observed in human CKD. For that, 6‐week‐old Wistar male rats were assigned to the two groups: the control group, 0.5% carboxymethyl cellulose (CMC) was administered; the CKD group, adenine was administered to rats at a dose 600 mg/kg for 10 days. The Raman spectra were obtained in the spectral range of 100–2500 cm−1. The excitation source was 532 nm, the laser power was ~40 mW, and the analyzed tissue area was 10.02 × 82.12 μm2. The results showed a significant reduction in the case of Raman peak intensities at 748, 1359, 1554, and 1636 cm−1, which can be assigned as Trp. Moreover, it was shown that tyrosine at 1169 cm−1 and phenylalanine at 1001 cm−1 decreased in the CKD group. Trp and kynurenine (Kyn) levels in kidney tissue and plasma were analyzed by using the high‐performance liquid chromatography (HPLC). The Trp concentrations in plasma and kidney tissue significantly decreased in CKD 7.4 μg/ml and 0.1 μg/mg, respectively. However, Kyn level was not a statistically significant change in kidney tissue. In this work, we showed that Raman spectroscopy usage in CKD model and Raman spectroscopy can be considered a promising technique for semi‐quantitative analysis of potential biomarkers such as phenylalanine, tyrosine, and Trp of renal function because of the little or no need for sample preparation, and it is easily observing several biomarkers at the same time. The utilization of multivariate analysis method, such as partial least square analysis can be considered as a promising tool for the discrimination of control and CKD groups.

Quantitative analysis of tryptophan and its metabolites in urine by ultra performance liquid chromatography-tandem mass spectrometry

基于超高效液相色谱-串联质谱(UPLC-MS/MS)建立定量分析色氨酸(Trp)及代谢产物3-OH-犬尿氨酸(3-OH-Kyn)、3-OH-邻氨基苯甲酸(3-OH-AA)、黄尿酸(XA)、犬尿氨酸(Kyn)、5-羟基吲哚乙酸(5-HIAA)、犬尿喹啉酸(KA)和5-羟色胺(5-HT)的方法,应用该方法分析其在尿样中的含量,探讨排泄规律。将尿样稀释、离心后,加入丹磺酰氯(DNS-Cl)衍生,经Thermo C18色谱柱(50 mm×3 mm, 2.7 μm)分离和0.1%甲酸和甲醇梯度洗脱后,采用电喷雾电离(ESI)源,在正离子扫描和多反应监测(MRM)模式下检测。以咖啡酸(CA)为内标,定量分析。结果显示,8种目标化合物的线性关系良好,相关系数(R 2)≥0.9740,检测灵敏(LOD为0.005~0.5 ng/mL),回收率高(93.24%~107.65%)。采用本方法检测分析了健康志愿者70个尿液样本,在尿样中检测到Trp原型及其7种代谢产物。结果表明,体内的Trp是通过原型和代谢两种方式排泄:Trp原型的含量为5.22~20.88 μg/mL;尿液中经代谢后排泄的Trp量是原型的124%~268%,即体内的Trp主要经代谢后排出体外。方法主要研究了Trp-5-HT和Trp-Kyn两条途径的代谢产物含量,Trp经Kyn降解生成的3-OH-AA和3-OH-Kyn含量较多,即Trp-Kyn是体内Trp的主要代谢途径。方法通过UPLC-MS/MS实现了尿液中Trp及其代谢产物含量的检测,能为临床检查提供技术和理论支持。

Estimation of dietary tryptophan requirement for laying duck breeders: effects on productive and reproductive performance, egg quality, reproductive organ and ovarian follicle development and serum biochemical indices

This study aimed to determine the dietary tryptophan (Trp) requirement for laying duck breeders. A total of 504 Longyan duck breeders (body weight: 1.20 ± 0.02 kg) aged 22 wk were randomly allocated to 6 treatments, each with 6 replicates of 14 ducks. During the next 16 wk, birds were fed the basal diet with total Trp contents of 1.00, 2.00, 3.00, 4.00, 5.00 and 6.00 g/kg, respectively. Dietary Trp levels increased egg production, egg mass and feed intake of duck breeders from 22 to 37 wk (P < 0.05), and there were linear and quadratic effects of Trp level (P < 0.05). The feed conversion ratio (FCR) quadratically decreased with dietary Trp levels (P < 0.05). Dietary Trp levels decreased (P < 0.05) egg albumen height and Haugh unit at wk 8 or 12, and the responses were linear and quadratic (P < 0.05). The body weight of breeders, absolute and relative weight of oviduct, number and total weight of preovulatory follicles (POF), and its proportion relative to ovarian weight were increased (P < 0.05), and the responses were linear (P < 0.01) and quadratic (P < 0.001). Ovarian weight increased quadratically (P < 0.05), and the mean weight of POF increased (P < 0.05), linearly and quadratically. The proportion of small yellow follicles relative to ovary weight decreased (P < 0.01) linearly and quadratically. At wk 16 of the trial period, the serum albumin content and alanine aminotransferase activity decreased (P < 0.05) and the creatinine content increased (P < 0.01) linearly and quadratically. The Trp requirements were estimated to be 3.14 g/kg for optimizing egg production, 2.93 g/kg for egg mass, and 2.92 g/kg for FCR. Overall, dietary Trp levels (1 to 6 g/kg) affected productive performance, egg quality, reproductive organ and ovarian follicle development, and serum biochemical indices of layer duck breeders, and a diet containing 2.9 to 3.1 g Trp per kg feed was adequate during the laying period (22 to 37 wk of age).

Hypoxia Routes Tryptophan Homeostasis Towards Increased Tryptamine Production.

The liver is the central hub for processing and maintaining homeostatic levels of dietary nutrients especially essential amino acids such as tryptophan (Trp). Trp is required not only to sustain protein synthesis but also as a precursor for the production of NAD, neurotransmitters and immunosuppressive metabolites. In light of these roles of Trp and its metabolic products, maintaining homeostatic levels of Trp is essential for health and well-being. The liver regulates global Trp supply by the immunosuppressive enzyme tryptophan-2,3-dioxygenase (TDO2), which degrades Trp down the kynurenine pathway (KP). In the current study, we show that isolated primary hepatocytes when exposed to hypoxic environments, extensively rewire their Trp metabolism by reducing constitutive Tdo2 expression and differentially regulating other Trp pathway enzymes and transporters. Mathematical modelling of Trp metabolism in liver cells under hypoxia predicted decreased flux through the KP while metabolic flux through the tryptamine branch significantly increased. In line, the model also revealed an increased accumulation of tryptamines under hypoxia, at the expense of kynurenines. Metabolic measurements in hypoxic hepatocytes confirmed the predicted reduction in KP metabolites as well as accumulation of tryptamine. Tdo2 expression in cultured primary hepatocytes was reduced upon hypoxia inducible factor (HIF) stabilisation by dimethyloxalylglycine (DMOG), demonstrating that HIFs are involved in the hypoxic downregulation of hepatic Tdo2. DMOG abrogated hepatic luciferase signals in Tdo2 reporter mice, indicating that HIF stability also recapitulates hypoxic rewiring of Trp metabolism in vivo. Also in WT mice HIF stabilization drove homeostatic Trp metabolism away from the KP towards enhanced tryptamine production, leading to enhanced levels of tryptamine in liver, serum and brain. As tryptamines are the most potent hallucinogens known, the observed upregulation of tryptamine in response to hypoxic exposure of hepatocytes may be involved in the generation of hallucinations occurring at high altitude. KP metabolites are known to activate the aryl hydrocarbon receptor (AHR). The AHR-activating properties of tryptamines may explain why immunosuppressive AHR activity is maintained under hypoxia despite downregulation of the KP. In summary our results identify hypoxia as an important factor controlling Trp metabolism in the liver with possible implications for immunosuppressive AHR activation and mental disturbances.

Standardized ileal digestible tryptophan to lysine ratios affect performance and regulate intestinal mRNA expression of amino acid transporters in weaning pigs fed a low crude protein diet

The objective of this study was to estimate the effect of standardized ileal digestible (SID) tryptophan (Trp) to lysine (Lys) ratios on growth performance and jejunal expression of amino acid transporters in weaning pigs. A total of 240 hybrid [(Duroc, male) x (Large White x Landrace, female)] piglets (6.52 ± 0.42 kg) were used in a 28-day growth trial and randomly allotted to one of four diets providing SID Trp to Lys ratios of 0.15, 0.18, 0.21, or 0.24 and were allocated in 6 pens per treatment with 10 pigs per pen. The average daily gain (ADG), average daily feed intake (ADFI), and serum levels of Trp, serine, isoleucine and leucine in pigs significantly increased when the SID Trp to Lys ratios increased from 0.15 to 0.18, 0.21, or 0.24 (P < 0.05). The pigs fed the diet with the 0.24 ratio had a reduced serum urea nitrogen (SUN) concentration compared with the pigs fed the 0.15 ratio diet (P < 0.05). The villus heights of the duodenum and jejunum in pigs fed the diet with a 0.21 ratio were greater than those of pigs fed the 0.15 ratio diet (P < 0.05). The increase in the SID Trp to Lys ratio from 0.15 to 0.18 decreased the intestinal ASCT2 mRNA level in weaning pigs (P < 0.05). The increase in the SID Trp to Lys ratio from 0.15 to 0.21 or 0.24 increased the mRNA abundance of intestinal B°AT1 (P < 0.05). The increase in the SID Trp to Lys ratio from 0.15 to 0.18 increased the mRNA abundances of intestinal CAT-1 and rBAT (P < 0.05). The intestinal b0,+AT mRNA level in the pigs fed the 0.21 ratio diet was higher than that in pigs in other three treatments (P < 0.05). The mRNA abundances of y+LAT1 and 4F2hc in the pigs fed the 0.15 ratio diet were lower than those in pigs in other three treatments (P < 0.05). The intestinal PepT-1 mRNA level in the pigs fed the 0.15 or 0.24 ratio diet was lower than that of the pigs fed the 0.21 ratio diet (P < 0.05). In conclusion, the optimal SID Trp to Lys ratio was determined to be 0.18 for weaning pigs that were fed a low crude protein diet using ADG and jejunal amino acid transporter mRNA expression as the response criteria.

Tryptophan metabolism is inversely regulated in the tumor and blood of patients with glioblastoma.

Tryptophan (Trp)-catabolic enzymes (TCEs) produce metabolites that activate the aryl hydrocarbon receptor (AHR) and promote tumor progression and immunosuppression in glioblastoma. As therapies targeting TCEs or AHR become available, a better understanding of Trp metabolism is required.Methods: The combination of LC-MS/MS with chemical isobaric labeling enabled the simultaneous quantitative comparison of Trp and its amino group-bearing metabolites in multiple samples. We applied this method to the sera of a cohort of 43 recurrent glioblastoma patients and 43 age- and sex-matched healthy controls. Tumor volumes were measured in MRI data using an artificial neural network-based approach. MALDI MSI visualized Trp and its direct metabolite N-formylkynurenine (FK) in glioblastoma tissue. Analysis of scRNA-seq data was used to detect the presence of Trp metabolism and AHR activity in different cell types in glioblastoma.Results: Compared to healthy controls, glioblastoma patients showed decreased serum Trp levels. Surprisingly, the levels of Trp metabolites were also reduced. The decrease became smaller with more enzymatic steps between Trp and its metabolites, suggesting that Trp availability controls the levels of its systemic metabolites. High tumor volume associated with low systemic metabolite levels and low systemic kynurenine levels associated with worse overall survival. MALDI MSI demonstrated heterogeneity of Trp catabolism across glioblastoma tissues. Analysis of scRNA-seq data revealed that genes involved in Trp metabolism were expressed in almost all the cell types in glioblastoma and that most cell types, in particular macrophages and T cells, exhibited AHR activation. Moreover, high AHR activity associated with reduced overall survival in the glioblastoma TCGA dataset.Conclusion: The novel techniques we developed could support the identification of patients that may benefit from therapies targeting TCEs or AHR activation.

High indoleamine-2,3-dioxygenase 1 (IDO) activity is linked to primary resistance to immunotherapy in non-small cell lung cancer (NSCLC).

BackgroundMetabolic profiling in non-small cell lung cancer (NSCLC) may identify key metabolic vulnerabilities and shows enormous discovery potential. Preclinical studies showed that metabolic rewiring in cancer plays an essential role in modulation of immunotherapy response. However, this situation is understudied in the clinical setting. Therefore, we aimed to evaluate the plasma metabolic profile of immune checkpoint inhibitor (CI) responding versus non-responding NSCLC patients. The aim of this project is to identify potential predictive biomarkers for CI response.MethodsPlasma samples from CI treated NSCLC patients were analysed at baseline and at the first follow up scan by using a broad targeted metabolomics mass spectrometry panel, and were compared to healthy controls. For further validation of identified key alterations high-performance liquid chromatography (HPLC) for tryptophan (Trp) and kynurenine (Kyn) as indicator of IDO-activity was performed.ResultsSixty-seven metabolites were significantly altered in NSCLC patients compared to healthy controls. The metabolic profile of patients with primary CI resistance showed an increase in indoleamine-2,3-dioxygenase (IDO) and a decrease in branched-chain amino acids (BCAA) compared to baseline concentrations. Deregulated IDO activity was validated by additional HPLC measurements, which revealed that baseline Trp levels were predictive for CI response. According to receiver operating characteristic (ROC)-analysis baseline Trp levels ≥49.3 µmol/L predicted disease control at the first follow up scan with a sensitivity of 89% and a specificity of 71%.ConclusionsWe showed that NSCLC patients are characterized by a distinct metabolic profile compared to healthy controls. Moreover, metabolic changes during CI therapy were observed. Of those IDO metabolism seemed to play an important role in primary CI resistance. Trp as a surrogate parameter of IDO activity is a promising biomarker in patients undergoing treatment with CIs and might be a future marker in trials investigating IDO inhibitors.

The Tryptophan System in Cocaine-Induced Depression

Major depression disorder (MDD) is the most prevalent psychiatric comorbid condition in cocaine use disorder (CUD). The comorbid MDD might be primary-MDD (CUD-primary-MDD) or cocaine-induced MDD (CUD-induced-MDD), and their accurate diagnoses and treatment is a challenge for improving prognoses. This study aimed to assess the tryptophan/serotonin (Trp/5-HT) system with the acute tryptophan depletion test (ATD), and the kynurenine pathway in subjects with CUD-primary-MDD, CUD-induced-MDD, MDD and healthy controls. The ATD was performed with a randomized, double-blind, crossover, and placebo-controlled design. Markers of enzymatic activity of indoleamine 2,3-dioxygenase/tryptophan 2,3-dioxygenase, kynurenine aminotransferase (KAT) and kynureninase were also established. Following ATD, we observed a decrease in Trp levels in all groups. Comparison between CUD-induced-MDD and MDD revealed significant differences in 5-HT plasma concentrations (512 + 332 ng/mL vs. 107 + 127 ng/mL, p = 0.039) and the Kyn/5-HT ratio (11 + 15 vs. 112 + 136; p = 0.012), whereas there were no differences between CUD-primary-MDD and MDD. Effect size coefficients show a gradient for all targeted markers (d range 0.72–1.67). Results suggest different pathogenesis for CUD-induced-MDD, with lower participation of the tryptophan system, probably more related to other neurotransmitter pathways and accordingly suggesting the need for a different pharmacological treatment approach.

The evaluation of serum tryptophan and kynurenine levels in patients with obstructive sleep apnea syndrome.

The kynurenine (Kyn) pathway may play a role in certain physiological functions such as behavior, sleep, thermoregulation, and pregnancy. Tryptophan (Trp) is oxidized with tryptophan 2,3-dioxygenase and indolamine 2,3-dioxygenase (IDO). Under normal conditions, hepatic kynurenine is a transcription factor and IDO expression in healthy tissues is very low. The ratio of Kyn to Trp can be used as an indicator to assess IDO activity. This study aimed to determine the relationship between Kyn/Trp ratio and obstructive sleep apnea syndrome (OSAS)disease activity. Study participants were categorized in 3 groups: Group 1 included patients withmild OSAS, Group 2, patients with moderatetosevere OSAS, and Group 3, individuals considered normalto serve as controls. The demographic characteristics of the patients were recorded. Apnea-hypopnea index (AHI) and oxygen desaturation index (ODI) measurements were performed by diagnostic polysomnography (PSG). Trp and Kyn levels were determined by HPLC-UV method. Group 1 included 30 patients (18 men) with mild OSAS;Group 2 included42 patients (31 men) with moderate to severe OSAS; and Group 3 included 25 controls (13 men).While there was no statistically significant difference between the levels of tryptophan and kynurenine in the groups, a significant difference was found between the Kyn/Trp ratios. A significant correlation was observed in individuals with a body mass index less than 25 with the Kyn/Trp ratio. In individuals with mild OSAS, a significant correlation was observed between ODI and BMI. In individuals with moderatetosevere OSAS, there was a significant correlation between ODI, AHI, and BMI. In this study, there was no relationship between OSAS diseaseseverityandIDO activity as assessed by immunoreactivity via the Kyn/Trp pathway.

Polydopamine/graphene/MnO2 composite-based electrochemical sensor for in situ determination of free tryptophan in plants

The in vivo detection of small active molecules in plant tissues is essential for the development of precision agriculture. Tryptophan (Trp) is an important precursor material for auxin biosynthesis in plants, and the detection of Trp levels in plants is critical for regulating the plant growth process. In this study, an electrochemical plant sensor was fabricated by electrochemically depositing a polydopamine (PDA)/reduced graphene oxide (RGO)-MnO2 nanocomposite onto a glassy carbon electrode (GCE). PDA/RGO-MnO2/GCE exhibited high electrocatalytic activity for the oxidation of Trp owing to the combined selectivity of PDA and catalytic activity of RGO-MnO2. To address the pH variability of plants, a reliable Trp detection program was proposed for selecting an appropriate quantitative detection model for the pH of the plant or plant tissue of interest. Therefore, a series of linear regression curves was constructed in the pH range of 4.0–7.0 using the PDA/RGO-MnO2/GCE-based sensor. In this pH range, the linear detection range of Trp was 1–300 μM, the sensitivity was 0.39–1.66 μA μM−1, and the detection limit was 0.22–0.39 μM. Moreover, the practical applicability of the PDA/RGO-MnO2/GCE-based sensor was successfully demonstrated by determining Trp in tomato fruit and juice. This sensor stably and reliably detected Trp levels in tomatoes in vitro and in vivo, demonstrating the feasibility of this research strategy for the development of electrochemical sensors for measurements in various plant tissues.

Effects of Dietary Crude Protein Levels on Fecal Crude Protein, Amino Acids Flow Amount, Fecal and Ileal Microbial Amino Acids Composition and Amino Acid Digestibility in Growing Pigs.

Simple SummaryThe purpose of this experiment was to evaluate a low protein corn-soybean meal-based diet with the same Lys, Met + Cys, Thr and Trp level as a high protein diet on fecal crude protein (CP), amino acid (AA) flow amount, AA digestibility and fecal and ileal microbial AA composition in growing pigs. Eighteen pigs with an initial body weight of (30 ± 1.35) kg were randomly divided into three groups, with six replicates in each group, and fed a corn-soybean meal-based diets with 12%, 15% and 18% CP levels, respectively. Our aim was to explain whether the addition of four crystalline essential AAs (EAA) to a low diet affected the digestibility of protein-bound NEAA (non-essential amino acid) and EAA and the composition of microbial AA in ileum and feces.The purpose of this experiment was to evaluate the effects of low protein corn-soybean meal-based diets on fecal CP, amino acid (AA) flow amount, AA digestibility and fecal and ileal microbial AA composition in growing pigs. Eighteen pigs (initial body weight = 30 ± 1.35) were randomly divided into three groups and fed with basal diets with CP levels of 12%, 15% and 18%, respectively. The Lys, Met + Cys, Thr and Trp level in the 12% CP and 15% CP groups is the same as 18% CP group by the addition of four crystalline Lys, Met + Cys, Thr and Trp to the diet. The results showed that with the decrease of dietary CP level from 18% to 12%, the fecal total nitrogen (N), CP and total AA (TAA) flow amount decreased linearly (p < 0.05). Dry matter (DM) digestibility, CP digestibility, TAA digestibility, essential amino acid (EAA) digestibility and non-essential amino acid (NEAA) digestibility increased linearly with the decrease of dietary CP concentration from 18% to 12%. Compared with 18% CP group, the flow amount of Asp, Ser, Glu, Gly, Tyr, Val, Leu and Phe in feces of pigs in the 15% CP group and 12% CP group decreased significantly, while the flow amount of Arg in the 15% CP group was lower than that in the 18% CP group and 12% CP group. The fecal microbial N and AA of the 15% CP group were higher than those of the 18% CP and 12% CP groups. Fecal TAA flow amount decreased linearly with the decrease of the dietary CP levels from 18% to 12%. Fecal TAA and NEAA flow amount also decreased linearly with the decrease of dietary CP level from 18% to 12%. Except for Glu, Gly, Met, Tyr, Thr and Phe, there were significant differences among the three groups in the composition of 17 kinds of AAs in fecal microorganisms. Among the 17 AA compositions of ileal microorganisms, except Tyr and Lys, the other AAs were significantly different among the three groups (p < 0.05)

Neopterin and biopterin levels and tryptophan degradation in patients with diabetes

This study aimed to evaluate the possible changes of neopterin, biopterin levels and tryptophan degradation in diabetes and to compare the results within diabetes groups and with healthy subjects. Diabetes mellitus patients and healthy controls were recruited the study. Patients were further subgrouped according to their drug therapy. Serum neopterin concentrations were detected by ELISA. Urinary neopterin, biopterin, serum tryptophan (Trp) and kynurenine (Kyn) levels were detected by HPLC. There was no difference between controls and diabetes patients in serum neopterin, urinary neopterin and biopterin levels (p > 0.05, all). Serum Trp and Kyn levels were significantly different in type 1 diabetes (T1DM) patients compared to controls (p < 0.05, both). Serum neopterin levels were significantly higher in type 2 diabetes patients (T2DM) compared to T1DM (p < 0.05). Urinary biopterin levels of T2DM patients using both metformin and vildagliptin were significantly higher than T1DM patients (p < 0.05). The correlations between serum neopterin and urinary neopterin, Kyn and Kyn/Trp were statistically significant in control and patient groups (p < 0.05, all). The study showed that Kyn/Trp was altered in diabetes patients due to immune modulation. On the other hand, although xenobiotic exposure may change pteridine levels, metformin and/or vildagliptin use in T2DM patients did not have any effect on the measured parameters.

Clinical significance of tryptophan catabolism in follicular lymphoma

The enzyme, indoleamine 2,3-dioxygenase 1 (IDO), catabolizes tryptophan (Trp) in the kynurenine (Kyn) pathway, and is important in suppressing antitumor immune responses in the tumor microenvironment. With regard to previously untreated patients with follicular lymphoma (FL), we sought to establish the prognostic significance of Trp catabolism in this disease. Serum Trp and Kyn levels in 110 patients with FL were quantified, and their relationship to different clinical parameters studied. IDO expression in the lymph nodes of affected patients was studied. Study participants included 54 males and 56 females (age range 39-86, median 62 years), showing a 5-year overall survival (OS) rate of 78.5%. Patients with a high Kyn level (5-year OS, 65.0% vs. 81.7%; p=0.026), high Kyn/Trp ratio (71.1% vs. 81.7%; p=0.002), and low hemoglobin (Hb) level (<12.0 g/dL; p=0.001; a component of FL international prognostic indexes) demonstrated a significantly shorter OS. Multivariate analysis included the following 10 variables: age, sex, serum β2-microglobulin, Hb, longest diameter of the largest involved node, Ann Arbor stage, serum lactate dehydrogenase, histologic grading, B symptoms, and serum Kyn/Trp ratio; a lower Hb level and a high Kyn/Trp ratio (HR, 3.239; 95% CI, 1.296-8.096) led to a significantly inferior OS. In the microenvironment, some CD11c-positive myeloid dendritic cells but not FL tumor cells were found to produce IDO. Overall, measuring levels of serum Kyn and Trp in individual patients with FL contributed to predicting their prognosis.

Association between IDO activity and prognosis in patients with non-small cell lung cancer after radiotherapy.

BackgroundIndoleamine 2,3-dioxygenase (IDO), a limiting enzyme in the IDO/kynurenine (Kyn) pathway, converts tryptophan (Trp) into Kyn, and plays a significant role in immune suppression and tumor immune evasion. This study aimed to investigate the association between IDO activity and clinical outcomes in non-small cell lung cancer (NSCLC) patients who underwent radiotherapy (RT).MethodsSerum Kyn and Trp levels were measured in 104 NSCLC patients by high-performance liquid chromatography at baseline, and the following RT. The correlation between IDO activity, as computed by Kyn: Trp ratios and survival was estimated using Kaplan-Meier curves. Cox proportional hazard models are used in the univariate and multivariate analyses.ResultsBoth the Kyn levels and Kyn:Trp ratios were reduced after RT at a biologically equivalent dose (BED) of <70 Gy, while these increased at a BED of ≥70 Gy. Post/pre-Kyn levels were positively correlated with an objective response. Patients with a higher Kyn:Trp ratio pre-RT had the worse median progression-free survival (mPFS, 13.5 vs. 24.5 months, P=0.049). Higher post/pre-Kyn:Trp ratios were correlated with improved median overall survival (mOS, 23.8 months vs. not reached, P=0.032). On the multivariate analysis, pre-RT Kyn:Trp and post/pre-Kyn:Trp ratios remained as independent predictive factors for PFS and OS, respectively.ConclusionsIt was proved that RT could alter IDO-mediated immune activity and establish strong correlations between IDO activity and survival outcomes in NSCLC patients treated with RT. These present findings suggest that the profiling of IDO activity might allow for the prompt adjustment of RT doses and better predict patient response to RT.

Standardized ileal digestible tryptophan and lysine for 15–30 kg gilts

AbstractIt was aimed to simultaneously study standardized ileal digestible (SID) tryptophan (Trp) and lysine (Lys) for gilts. A digestibility assay was previously conducted to determine the SID amino acid in the basal diet (low levels of SID Trp and Lys). Sixty-four gilts (15.04 ± 1.44 kg) were allotted to 16 diets in a 4 × 4 factorial arrangement (1.55, 1.85, 2.15 and 2.45 g/kg SID Trp and 9.72, 11.12, 12.52 and 13.92 g/kg SID Lys) with four replicates per treatment. Performance, longissimus muscle (LM), backfat thickness (BF) and blood variables were evaluated. An interaction was observed for G:F, and by response surface model, the optimum Trp level was achieved at 2.15 g/kg (0.159 g/MJ of ME). A quadratic effect of Trp was observed on body weight (BW) and average daily gain (ADG); the daily feed intake increased linearly as Trp increased. The optimum Trp levels of 2.25 and 2.24 g/kg were estimated for BW and ADG, respectively. The BF increased with increasing levels of Trp. There was a quadratic and linear effect of Trp and Lys, respectively, on the LM, in which the optimum Trp level was determined as 2.05 g/kg in the diet. Plasma urea nitrogen decreased as Trp and Lys levels increased. Using estimates provided by response surface, maximized G:F ratio was obtained at 2.15 g SID Trp/kg of diet and at least 13.92 g SID Lys/kg of diet is necessary to optimize the G:F for 15–30 kg gilts, providing a Trp:Lys ratio of 15.4:100.