Trp Concentrations Research Articles

Dietary tryptophan alters CD4+ T cell function in the lupus-prone B6. Sle1.Sle2.Sle3 (TC) mouse model

Abstract Tryptophan (Trp) metabolism is altered in lupus patients with higher production of kynurenine (Kyn). We show that TC mice have the same alterations. To ascertain whether Trp plays a role in autoimmune pathogenesis, TC and B6 control mice were fed with either high or low Trp. High Trp exacerbated disease in TC mice while low Trp was protective. Expression of IDO1, the enzyme producing Kyn from Trp, was similar between TC and B6 mice, and its inhibition by 1MT did not change disease outcomes with and without dietary Trp variations. To identify the mechanism by which Trp plays a role in this process, we analyzed CD4 T cell phenotypes. Regulatory T cell (Treg) functional markers were altered by dietary Trp. Further, Tregs from TC mice consuming high Trp lost their suppressive capacity in vitro, whereas Tregs from TC mice on low Trp were as suppressive as that of B6 controls. Additionally, increasing Trp concentrations affected B6 and TC T cell polarization in vitro differently. A high Trp concentration increased the percentage of IFNg+ and FoxP3+ CD4+ T cells from B6 and TC mice under Th1 and Treg polarizing conditions, respectively. On the other hand, B6 Treg polarization increased with a higher Kyn concentration while TC Treg polarization decreased. These findings suggest that Trp and some of its metabolites modulate lupus pathogenesis in TC mice potentially by altering T cell phenotypes. Thus, these data emphasize a novel pro-inflammatory role for Trp metabolism in lupus. Supported by a grant from the NIH (RO1 AI143313) to LM.

Zeolitic imidazolate frameworks and cobalt-tannic acid nanocomposite modified carbon paste electrode for simultaneous determination of dopamine, uric acid, acetaminophen and tryptophan: Investigation of kinetic parameters of surface electrode and its analytical performance

In this study, we have created a particularly selective electrochemical sensor cobalt-tannic acid nanocomposite (ZIF-8@Co-TA) and a familiar metal organic framework (ZIF-8), which was then used to simultaneously determine acetaminophen (AC), dopamine (DA), tryptophan (Trp), and uric acid (UA) with wide linear ranges and low detection limits (LOD). In addition, we have applied chronoamperometry, differential pulse voltammetry (DPV), electrochemical impedance spectroscopy, and cyclic voltammetry so that the electrochemical features of the fabricated electrode could be assessed. The eminent catalytic activity and high-level conductivity of ZIF-8@Co-TA has led to perfectly separated peaks, and to improved peak currents of DA, UA, AC, and Trp. According to the results, the ΔEp of 130, 120, 310 mV vs. SCE were reported as the level of separation of anodic peak potentials for DA-UA, UA-AC, and AC-Trp, respectively. With LOD of 3.4, 1.2, 5.1 and 6.7 nM, the maximum currents linearly depended on DA, UA, AC and Trp concentrations in the ranges of 0.02–0.44 μM (Sbk/m = 3). Good repeatability and stability were observed after determining DA, UA, AC, and Trp, which was carried out through successfully challenging the chemically modified electrode with some interfering compounds and evaluating it in real samples. Therefore, it seems that fabricated sensor can be properly exploited for clinical experiments and pharmaceutical applications.

Application of carboxylic acid-functionalized of graphene oxide for electrochemical simultaneous determination of tryptophan and tyrosine in milk

In this work, a new method for the simultaneous determination of tryptophan (Trp) and tyrosine (Tyr) in milk has been reported. Trp and Tyr are essential amino acids in human nutrition, but their electrochemical signal overlapped. For solve this problem, we developed modified Screen Printed Carbon Electrode (SPCE) with Graphene Oxide–COOH/Chitosan (GO–COOH/Chitosan) electro-deposition. The morphology and electrochemical performance of modified electrode were characterized by scanning electron microscopy, energy dispersive spectrometry, FT-IR, electrochemical impedance spectroscopy and cyclic voltammetry. The electrochemical simultaneous determination of Trp and Tyr has been investigated by using differential pulse voltammetry. The carboxylic acid functionalized GO modified SPCE was utilized to catalyze the oxidation of Trp and Tyr. Compared with SPCE/GO/Chitosan and SPCE/Chitosan sensor, the new sensor has enhanced sensitivity, low detection limit and high selectivity. In addition, the SPCE/GO–COOH–Chitosan sensor enhanced separation of the oxidation peak of Tyr and Trp and showed a remarkable increase in peak current for electro-active compounds, thus, it can be used for simultaneous voltammetric determination. Under the optimized experimental conditions, a linear correlation between oxidation peak current and concentration of Tyr and Trp in the ranges 0.1–60 µM and 0.4–40 µM with a detection limit of 0.05 µM (S/N = 3) and 0.1 µM (S/N = 3) were achieved, respectively. Finally, the proposed electrochemical sensor was applied for quantification of Tyr and Trp in milk samples, where standard solutions were spiked to the milk samples and recoveries were obtained.

Serum Levels and Removal by Haemodialysis and Haemodiafiltration of Tryptophan-Derived Uremic Toxins in ESKD Patients.

Tryptophan is an essential dietary amino acid that originates uremic toxins that contribute to end-stage kidney disease (ESKD) patient outcomes. We evaluated serum levels and removal during haemodialysis and haemodiafiltration of tryptophan and tryptophan-derived uremic toxins, indoxyl sulfate (IS) and indole acetic acid (IAA), in ESKD patients in different dialysis treatment settings. This prospective multicentre study in four European dialysis centres enrolled 78 patients with ESKD. Blood and spent dialysate samples obtained during dialysis were analysed with high-performance liquid chromatography to assess uremic solutes, their reduction ratio (RR) and total removed solute (TRS). Mean free serum tryptophan and IS concentrations increased, and concentration of IAA decreased over pre-dialysis levels (67%, 49%, −0.8%, respectively) during the first hour of dialysis. While mean serum total urea, IS and IAA concentrations decreased during dialysis (−72%, −39%, −43%, respectively), serum tryptophan levels increased, resulting in negative RR (−8%) towards the end of the dialysis session (p < 0.001), despite remarkable Trp losses in dialysate. RR and TRS values based on serum (total, free) and dialysate solute concentrations were lower for conventional low-flux dialysis (p < 0.001). High-efficiency haemodiafiltration resulted in 80% higher Trp losses than conventional low-flux dialysis, despite similar neutral Trp RR values. In conclusion, serum Trp concentrations and RR behave differently from uremic solutes IS, IAA and urea and Trp RR did not reflect dialysis Trp losses. Conventional low-flux dialysis may not adequately clear Trp-related uremic toxins while high efficiency haemodiafiltration increased Trp losses.

Formation and characterization of crosslinks, including Tyr-Trp species, on one electron oxidation of free Tyr and Trp residues by carbonate radical anion.

Dityrosine and ditryptophan bonds have been implied in protein crosslinking. This is associated with oxidative stress conditions including those involved in neurodegenerative pathologies and age-related processes. Formation of dityrosine and ditryptophan derives from radical–radical reactions involving Tyr˙ and Trp˙ radicals. However, cross reactions of Tyr˙ and Trp˙ leading to Tyr–Trp crosslinks and their biological consequences have been less explored. In the present work we hypothesized that exposure of free Tyr and Trp to a high concentration of carbonate anion radicals (CO3˙−), under anaerobic conditions, would result in the formation of Tyr–Trp species, as well as dityrosine and ditryptophan crosslinks. Here we report a simple experimental procedure, employing CO3˙− generated photochemically by illumination of a Co(iii) complex at 254 nm, that produces micromolar concentrations of Tyr–Trp crosslinks. Analysis by mass spectrometry of solutions containing only the individual amino acids, and the Co(iii) complex, provided evidence for the formation of o,o′-dityrosine and isodityrosine from Tyr, and three ditryptophan dimers from Trp. When mixtures of Tyr and Trp were illuminated in an identical manner, Tyr–Trp crosslinks were detected together with dityrosine and ditryptophan dimers. These results indicate that there is a balance between the formation of these three classes of crosslinks, which is dependent on the Tyr and Trp concentrations. The methods reported here allow the generation of significant yields of isolated Tyr–Trp adducts and their characterization. This technology should facilitate the detection, and examination of the biological consequences of Tyr–Trp crosslink formation in complex systems in future investigations.

Up-regulation of indoleamine 2,3-dioxygenase 1 (IDO1) expression and catalytic activity is associated with immunosuppression and poor prognosis in penile squamous cell carcinoma patients.

BackgroundIndoleamine 2,3‐dioxygenase 1 (IDO1) and tryptophan (Trp) catabolism have been demonstrated to play an important role in tumor immunosuppression. This study examined the expression and catalytic activity of IDO1 in penile squamous cell carcinoma (PSCC) and explored their clinical significance.MethodsIDO1 expression level, serum concentrations of Trp and kynurenine (Kyn) were examined in 114 PSCC patients by immunohistonchemistry and solid‐phase extraction‐liquid chromatography‐tandem mass spectrometry. The survival was analyzed using Kaplan‐Meier method and the log‐rank test. Hazard ratio of death was analyzed via univariate and multivariate Cox regression. Immune cell types were defined by principal component analysis. The correlativity was assessed by Pearson's correlation analysis.ResultsThe expression level of IDO1 in PSCC cells was positively correlated with serum Kyn concentration and Kyn/Trp radio (KTR; both P < 0.001) but negatively correlated with serum Trp concentration (P = 0.001). Additionally, IDO1 up‐regulation in cancer cells and the increase of serum KTR were significantly associated with advanced N stage (both P < 0.001) and high pathologic grade (P = 0.008 and 0.032, respectively). High expression level of IDO1 in cancer cells and serum KTR were associated with short disease‐specific survival (both P < 0.001). However, besides N stage (hazard radio [HR], 6.926; 95% confidence interval [CI], 2.458‐19.068; P < 0.001) and pathologic grade (HR, 2.194; 95% CI, 1.021‐4.529; P = 0.038), only serum KTR (HR, 2.780; 95% CI, 1.066‐7.215; P = 0.036) was an independent predictor for PSCC prognosis. IDO1 expression was positively correlated with the expression of interferon‐γ (IFNγ, P < 0.001) and immunosuppressive markers (programmed cell death protein 1, cytotoxic T‐lymphocyte‐associated protein 4 and programmed death‐ligand 1 and 2; all P < 0.05), and the infiltration of immune cells (including cytotoxic T lymphocytes, regulatory T lymphocytes, tumor‐associated macrophages, and myeloid‐derived suppressor cells; all P < 0.001) in PSCC tissues. Furthermore, the expression of IDO1 was induced by IFNγ in a dose‐dependent manner in PSCC cells.ConclusionsIFNγ‐induced IDO1 plays a crucial role in immunoediting and immunosuppression in PSCC. Additionally, serum KTR, an indicator of IDO1 catabolic activity, can be utilized as an independent prognostic factor for PSCC.

Detection of a High-Turnover Serotonin Circuit in the Mouse Brain Using Mass Spectrometry Imaging.

SummaryMonoamine neurotransmitters are released by specialized neurons regulating behavioral, motor, and cognitive functions. Although the localization of monoaminergic neurons in the brain is well known, the distribution and kinetics of monoamines remain unclear. Here, we generated a murine brain atlas of serotonin (5-HT), dopamine (DA), and norepinephrine (NE) levels using mass spectrometry imaging (MSI). We found several nuclei rich in both 5-HT and a catecholamine (DA or NE) and identified the paraventricular nucleus of the thalamus (PVT), where 5-HT and NE are co-localized. The analysis of 5-HT fluctuations in response to acute tryptophan depletion and infusion of isotope-labeled tryptophan in vivo revealed a close kinetic association between the raphe nuclei, PVT, and amygdala but not the other nuclei. Our findings imply the existence of a highly dynamic 5-HT-mediated raphe to PVT pathway that likely plays a role in the brain monoamine system.

Rapid recognition and determination of tryptophan by carbon nanotubes and molecularly imprinted polymer-modified glassy carbon electrode

A tryptophan (Trp) molecularly imprinted electrochemical sensor was fabricated by drop-coating an imprinted chitosan film on the surface of a glassy carbon electrode modified with multi-walled carbon nanotubes (MIP-MWCNTs/GCE). The surface morphology and electrochemical properties of the MIP-MWCNTs/GCE were characterized by scanning electron microscopy (SEM) and cyclic voltammetry (CV), respectively. The formation of hydrogen bonds between the functional polymer and the template molecule was confirmed by infrared spectroscopy. The electrochemical performance of the MIP-MWCNTs/GCE with Trp showed that the signal of the oxidation current of Trp obtained with MIP-MWCNTs/GCE was significantly enhanced relative to that of the uncovered GCE, indicating that the modified electrode can accelerate electron transfer and has strong selectivity for Trp. The experimental conditions were optimized in parallel, and under the optimal conditions, the MIP-MWCNTs/GCE showed a good linear relationship between the Trp oxidation peak current and Trp concentrations in the ranges of 2.0 nM–0.2 μM, 0.2 μM–10 μM and 10 μM–100 μM The limit of detection (LOD) was 1.0 nM (S/N = 3), and the modified electrode had good reproducibility and stability. Finally, the MIP-MWCNTs/GCE was successfully applied to the determination of Trp in the human serum samples.

Development of an effective sample transfer device for biomarker detection in nasal secretions

Nasal secretions (NS) reflect inflammatory activity of the nasal mucosa and thus can be utilized for disease diagnosis and determining treatment effects in Allergic rhinitis (AR). However, non-standardized collection of samples can affect the measured concentration of inflammatory biomarker in NS. In this study, we aimed to develop and evaluate new devices capable of standardizing the collection, storage, and preprocessing methods of NS samples. First, we chose the best swab as polyester (PE) and selected a stimulation method, twirling for 10 s at 1 Hz, to efficiently release AR biomarkers from a PE swab. Storage of sample solutions at −20 °C was optimal for the stability of biomarkers for the detection of AR. The new swab sample transfer device showed excellent concentration recovery efficiency (90–100%) for tryptase (Trp) and eosinophil cationic protein (ECP) without crosstalk between the two biomarkers. Finally, we compared the concentration of Trp in human NS samples of AR patients (n = 6) pre-processed by the new device with that by centrifuge as a standard method. As a result, the concentrations of Trp in NS were very similar in both groups. Therefore, this device can be utilized as an effective sample transfer and pre-processing device for point-of-care testing of AR.

Application of the slope‐ratio growth assay technique to estimate tryptophan availability in soybean meal fed to young rats

The slope-ratio assay for rat was used to determine whether tryptophan (Trp) availability in soybean meal (SBM) is affected by the presence of other amino acids (AAs). In a preliminary study, rats were fed graded levels of Trp-supplemented diets to establish the Trp concentration range over which the weight gain response was linear. This range was found to be from 0.04% to 0.12% Trp. Subsequently, rats were fed basal (0.045% Trp) or Trp-supplemented diets from three different sources: l-Trp alone, SBM, or l-Trp mixed with other AAs to reflect AA levels in the test SBM (AA-mix). Weight gain in rats increased linearly with supplemental Trp intake (p<.05) for all Trp sources. Compared to the slope achieved with l-Trp alone, the estimated availability of Trp in SBM was 84.4%, while for the AA-mix it was 93.4%. It is evident that the 6.6% reduction in l-Trp availability in AA-mix is due to metabolic costs derived from excess levels of other AAs beside Trp, given that the absorption of crystalline l-Trp in the small intestine is 100%. In conclusion, the Trp availability of SBM was estimated to be around 90.4% (i.e., 84.4/93.4×100) after correcting for the effects of the other AAs in SBM.

Metabolic and Stress Responses in Senegalese Soles (Solea senegalensis Kaup) Fed Tryptophan Supplements: Effects of Concentration and Feeding Period.

Simple SummaryIn order to improve fish welfare in fish farms, feed additives (tryptophan, Trp) were assayed in a cultured species, the Senegalese sole (Solea senegalensis). It has been observed in previous studies that fish fed Trp supplements for several days prior to a stress challenge improved their physiological stress response. Therefore, this feeding strategy could be applied in commercial fish farms before submitting fish to stressful zootechnical procedures, such as grading, sampling, slaughter, and, consequently, fish welfare would be improved.The objective of this study was to assess the impact of different dietary Trp concentrations on the stress and metabolism response of juvenile Senegalese soles (Solea senegalensis). Fish (38.1 ± 1.9 g) were fed different Trp-enriched feeds (0%, 1% and 2% Trp added) for two and eight days, and later exposed to air stress for three min. Samples were taken pre- and 1 h post-stress (condition). Plasma cortisol, lactate, glucose and proteins were significantly affected by the sampling time, showing higher values at 1 h post-stress. Trp concentration in food also had significant effects on lactate and glucose levels. However, the feeding period did not affect these parameters. Post-stress values were higher than in the pre-stress condition for every plasma parameter, except for lactate in two days and 1% Trp treatment. Nevertheless, cortisol, glucose and lactate did not vary significantly between pre- and post-stress samplings in fish fed the 1% Trp-enriched diet for two days. The lack of variability in cortisol response was also due to the high pre-stress value, significantly superior to pre-stress control. The exposure time to Trp feeding did not significantly affect any enzyme activity; however, Trp added and condition influenced protein-related enzyme activities. In spite of decreasing stress markers, Trp-enriched diets altered the protein metabolism.

Electrochemical Sensor for Rapid and Sensitive Detection of Tryptophan by a Cu2O Nanoparticles-Coated Reduced Graphene Oxide Nanocomposite.

In this paper, a nanocomposite of cuprous oxide and electrochemically reduced graphene oxide (Cu2O–ERGO) was prepared by a simple and low-cost method; hereby, a new method for the electrochemical determination of tryptophan (Trp) by this composite modified glassy carbon electrode (GCE) is proposed. The prepared materials and modified electrodes were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and cyclic voltammetry (CV). The results showed that Cu2O–ERGO/GCE had good electrocatalytic activity for Trp. The effects of supporting electrolyte, scanning rate, accumulation potential, and accumulation time on the determination of Trp were studied. Under the optimum experimental conditions, Trp was quantitatively analyzed by square-wave voltammetry (SWV). The oxidation peak current of Trp had a good linear relationship with its concentration in the range of 0.02–20 μM, and the detection limit was 0.01 μM (S/N = 3). In addition, the modified electrode has high sensitivity, good repeatability, and long-term stability. Finally, the proposed method has been successfully applied in the determination of Trp concentration in practical samples.

Perylene-functionalized graphene sheets modified with chitosan for voltammetric discrimination of tryptophan enantiomers.

A composite was prepared from graphene functionalized with 3,4,9,10-perylene tetracarboxylic acid and chitosan (rGO-PTCA-chitosan) by a chemical method. It involves non-covalent functionalization of rGO with PTCA followed by amidation reaction with chitosan. Scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy and electrochemical methods were used to characterize the composites. By combining the chiral features of chitosan and the excellent electrochemical behaviors of rGO-PTCA, a graphene-based material with enantioselectivity was constructed for electrochemical chiral recognition of tryptophan (Trp) enantiomers. A glassy carbon electrode (GCE) modified with rGO-PTCA-chitosan had a higher recognition capability for L-Trp than for D-Trp. Best operated at a working voltage near 0.78 V (vs. SCE), the enantioselectivity coefficient is 3.0. The sensor has a linear response in the 1mM to 10mM Trp concentration range and a 1.2μM detection limit (at S/N= 3) for L-Trp, and of 3.0μM to D-Trp. The sensor was successfully used to detect Trp enantiomers in real samples, and a recognition mechanism is presented. Graphical abstract Schematic presentation of a composoie prepared by graphene functionalized with 3,4,9,10-perylene tetracarboxylic acid and chitosan (rGO-PTCA-chitosan) via a chemical method. It involves non-covalent functionalization of rGO with PTCA followed by amidation reaction with chitosan and voltammetric determination of tryptophan enantiomers.

CRISPR/Cas9-facilitated engineering with growth-coupled and sensor-guided in vivo screening of enzyme variants for a more efficient chorismate pathway in E. coli

Protein engineering plays an increasingly important role in developing new and optimizing existing metabolic pathways for biosynthesis. Conventional screening approach of libraries of gene and enzyme variants is often done using a host strain under conditions not relevant to the cultivation or intracellular conditions of the later production strain. This does not necessarily result in the identification of the best enzyme variant for in vivo use in the production strain. In this work, we propose a method which integrates CRISPR/Cas9-facilitated engineering of the target gene(s) with growth-coupled and sensor-guided in vivo screening (CGSS) for protein engineering and pathway optimization. The efficiency of the method is demonstrated for engineering 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase AroG, a key enzyme in the chorismate pathway for the synthesis of aromatic amino acids (AAAs), to obtain variants of AroG (AroGfbr) with increased resistance to feedback inhibition of Phe. Starting from a tryptophan (Trp)-producing E. coli strain (harboring a reported Phe-resistant AroG variant AroGS180F), the removal of all the endogenous DAHP synthases makes the growth of this strain dependent on the activity of an introduced AroG variant. The different catalytic efficiencies of AroG variants lead to different intracellular concentration of Trp which is sensed by a Trp biosensor (TnaC-eGFP). Using the growth rate and the signal strength of the biosensor as criteria, we successfully identified several novel Phe-resistant AroG variants (including the best one AroGD6G−D7A) which exhibited higher specific enzyme activity than that of the reference variant AroGS180F at the presence of 40 mM Phe. The replacement of AroGS180F with the newly identified AroGD6G−D7A in the Trp-producing strain significantly improved the Trp production by 38.5% (24.03 ± 1.02 g/L at 36 h) in a simple fed-batch fermentation.

Plasma indoleamine 2,3-dioxygenase activity as a potential biomarker for early diagnosis of multidrug-resistant tuberculosis in tuberculosis patients.

Purpose: Multidrug-resistant tuberculosis (MDR-TB) remains a challenge of global TB control, with difficulty in early detection of drug-sensitive tuberculosis (DS-TB). We investigate the diagnostic significance of IDO as a potential biomarker to discriminate MDR patients among the TB patients.Patients and methods: Plasma indoleamine 2,3-dioxygenase (IDO) was measured by the ratio of kynurenine (Kyn) to tryptophan (Trp) concentrations, using high performance liquid chromatography-mass spectrometry (LC-MS/MS). Chest computed tomography (CT) imaging signs from TB patients were collected and analyzed in 18 DS-TB patients, 16 MDR-TB patients, 6 lung cancer (LC) patients, and 11 healthy individuals. Lung imaging signs from TB patients were collected and analyzed.Results: We found that plasma IDO activity was significantly higher in the MDR-TB patients than in the DS-TB patients (p=0.012) and in the LC patients (p=0.003). We evaluated the diagnostic significance of plasma IDO activity in discriminating the MDR-TB group from the DS-TB group using a receiver operating characteristic (ROC) curve. With a cutoff level of 46.58 uM/mM, the diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for IDO activity were 87.50%, 72.22%, 73.68%, and 86.67%, respectively. Plasma IDO activity was higher in cavity cases than in non-cavity cases (p=0.042), proving a positive correlation between lung cavity number and cavity size (p<0.05, separately) among all the TB patients studied.Conclusion: Our findings confirmed that plasma IDO activity might have an auxiliary diagnosis value for early discrimination of MDR-TB patients from DS-TB patients. Among the TB patients with cavitary lung lesions, higher plasma IDO activity can indicate a higher risk of MDR-TB.

Tryptophan Metabolic Pathways and Brain Serotonergic Activity: A Comparative Review.

The essential amino acid L-tryptophan (Trp) is the precursor of the monoaminergic neurotransmitter serotonin (5-hydroxytryptamine, 5-HT). Numerous studies have shown that elevated dietary Trp has a suppressive effect on aggressive behavior and post-stress plasma cortisol concentrations in vertebrates, including teleosts. These effects are believed to be mediated by the brain serotonergic system, even though all mechanisms involved are not well understood. The rate of 5-HT biosynthesis is limited by Trp availability, but only in neurons of the hindbrain raphe area predominantly expressing the isoform TPH2 of the enzyme tryptophan hydroxylase (TPH). In the periphery as well as in brain areas expressing TPH1, 5-HT synthesis is probably not restricted by Trp availability. Moreover, there are factors affecting Trp influx to the brain. Among those are acute stress, which, in contrast to long-term stress, may result in an increase in brain Trp availability. The mechanisms behind this stress induced increase in brain Trp concentration are not fully understood but sympathetic activation is likely to play an important role. Studies in mammals show that only a minor fraction of Trp is utilized for 5-HT synthesis whereas a larger fraction of the Trp pool enters the kynurenic pathway. The first stage of this pathway is catalyzed by the hepatic enzyme tryptophan 2,3-dioxygenase (TDO) and the extrahepatic enzyme indoleamine 2,3-dioxygenase (IDO), enzymes that are induced by glucocorticoids and pro-inflammatory cytokines, respectively. Thus, chronic stress and infections can shunt available Trp toward the kynurenic pathway and thereby lower 5-HT synthesis. In accordance with this, dietary fatty acids affecting the pro-inflammatory cytokines has been suggested to affect metabolic fate of Trp. While TDO seems to be conserved by evolution in the vertebrate linage, earlier studies suggested that IDO was only present mammals. However, recent phylogenic studies show that IDO paralogues are present within the whole vertebrate linage, however, their involvement in the immune and stress reaction in teleost fishes remains to be investigated. In this review we summarize the results from previous studies on the effects of dietary Trp supplementation on behavior and neuroendocrinology, focusing on possible mechanisms involved in mediating these effects.

Activation of the kynurenine pathway is associated with poor outcome in Pneumocystis pneumonia patients infected with HIV: results of 2\u2009months cohort study

BackgroundIndoleamine 2, 3-dioxygenase (IDO) is a key enzyme in the degradation of tryptophan (Trp) to kynurenine (Kyn). We measured IDO activity as the Kyn to Trp ratio, and investigated whether IDO could be used to assess prognosis of acquired immune deficiency Sydrome (AIDS) patients with pneumocystis pneumonia (PCP).MethodsThe Kyn and Trp concentration were measured by UPLC-MS/MS in plasma samples. A total of 49 AIDS-PCP patients were included in the analysis. Clinical characteristics and Kyn/Trp ratio were compared between survivors and non-survivors.ResultsKyn/Trp ratio was significantly lower after anti-PCP treatment in AIDS patients with PCP (P < 0.0001). Plasma Kyn/Trp ratio was higher in patients with PaO2/FiO2 ≤ 300 mmHg than in those with PaO2/FiO2 > 300 mmHg (P = 0.007). Kyn/Trp ratio, D-dimer and CRP showed much higher AUC for predicting death of AIDS-PCP patients. Kyn/Trp ratio was useful for predicting the mortality of AIDS-PCP due to a significantly higher Kyn/Trp ratio in the non-survivors (P = 0.002). And the high Kyn/Trp ratio group had higher mortality rate than low Kyn/Trp group (32.1% vs. 9.1%, respectively, p = 0.024).ConclusionActivation of the kynurenine pathway is associated with the severity and fatal outcomes of AIDS patients with pneumocystis pneumonia.

Evaluation of the tryptophan requirement of small-framed first cycle laying hens

An experiment was conducted to evaluate the total Trp requirement of first cycle laying hens from 41 to 60 wk of age. In total, 270 Hy-Line W36 laying hens were randomly allocated to six treatments with 15 replicate groups of three birds for each experimental unit. A Trp-deficient basal diet was formulated with corn, corn gluten meal, and soybean meal with determined Trp, Lys, and crude protein concentrations of 0.096%, 0.873%, and 15.0%, respectively. Synthetic L-Trp was supplemented to the basal diet in 0.020% increments to generate experimental diets containing 0.096%, 0.116%, 0.136%, 0.156%, 0.176%, and 0.196% Trp, respectively. Hens were provided a controlled amount of feed of approximately 95g/d. The diet containing the lowest concentration of Trp resulted in reduced egg production and was halted at 45 wk of age due to low performance with all other dietary treatments reaching the conclusion of the experiment at 60 wk of age. Plasma serotonin responded to dietary Trp concentration, but was not a good candidate for Trp requirement estimation. Linear broken line, quadratic broken line, quadratic polynomial, and exponential models were used to estimate Trp requirement based on hen-housed egg production (HHEP), egg mass (EM), and feed conversion ratio (FCR). HHEP ranged from 50.7% to 81.0%, dependent upon dietary concentration of Trp. Tryptophan requirements estimated by linear broken line, quadratic broken line, quadratic polynomial, and exponential models were reported. Using the linear broken line model, Trp requirement was highest for EM, 155.8mg/d, followed by egg production, 153.2mg/d and lowest for FCR, 140.4mg/d.

Kynurenine Is a Cerebrospinal Fluid Biomarker for Bacterial and Viral Central Nervous System Infections.

The tryptophan-kynurenine-nicotinamide adenine dinucleotide (oxidized; NAD+) pathway is closely associated with regulation of immune cells toward less inflammatory phenotypes and may exert neuroprotective effects. Investigating its regulation in central nervous system (CNS) infections would improve our understanding of pathophysiology and end-organ damage, and, furthermore, open doors to its evaluation as a source of diagnostic and/or prognostic biomarkers. We measured concentrations of kynurenine (Kyn) and tryptophan (Trp) in 221 cerebrospinal fluid samples from patients with bacterial and viral (due to herpes simplex, varicella zoster, and enteroviruses) meningitis/encephalitis, neuroborreliosis, autoimmune neuroinflammation (due to anti-N-methyl-D-aspartate receptor [NMDA] encephalitis and multiple sclerosis), and noninflamed controls (ie, individuals with Bell palsy, normal pressure hydrocephalus, or Tourette syndrome). Kyn concentrations correlated strongly with CSF markers of neuroinflammation (ie, leukocyte count, lactate concentration, and blood-CSF-barrier dysfunction), were highly increased in bacterial and viral CNS infections, but were low or undetectable in NMDA encephalitis, multiple sclerosis, and controls. Trp concentrations were decreased mostly in viral CNS infections and neuroborreliosis. Multiple logistic regression analysis revealed that combinations of Kyn concentration, Trp concentration, and Kyn/Trp concentration ratio with leukocyte count or lactate concentration were accurate classifiers for the clinically important differentiation between neuroborreliosis, viral CNS infections, and autoimmune neuroinflammation. The Trp-Kyn-NAD+ pathway is activated in CNS infections and provides highly accurate CSF biomarkers, particularly when combined with standard CSF indices of neuroinflammation.

Ultrasensitive and reusable electrochemical aptasensor for detection of tryptophan using of [Fe(bpy)3](p-CH3C6H4SO2)2 as an electroactive indicator

In this paper, we report the application of a reusable electrochemical aptasensor for detection of tryptophan by using [Fe(bpy)3](p-CH3C6H4SO2)2 as an electroactive indicator and based on the target-compelled aptamer displacement. The aptasensor fabricated by self-assembling the thiolated probe on the surface of graphite screen-printed electrode modified with gold nanoparticles/multiwalled carbon nanotubes and chitosan nanocomposite (AuNPs/MWCNTs-Chit/SPE). Afterward, Trp aptamer (Apt) immobilized on the modified electrode surface through hybridization. In the absence of Trp, a sharp peak of [Fe(bpy)3](p-CH3C6H4 SO2)2 can be observed in differential pulse voltammetry (DPV) study. The introduction of Trp led to the formation of aptamer-Trp complex and dissociation of the aptamer from the DNA-Apt duplex on the electrode surface into the solution and decreases the peak current intensity of electroactive indicator. This is because, [Fe(bpy)3](p-CH3C6H4SO2)2 tends to bind to the two strands DNA. Therefore, the peak current of [Fe(bpy)3](p-CH3C6H4 SO2)2 linearly decreased with increasing the concentration of Trp over a range of 3.0 nM- 100 μM. The detection limit (3 σ) was 1.0 nM. In addition, we examined the selectivity of the constructed biosensor for tyrosine, histidine, arginine, lysine, valine and methionine that belonged to the amino acid family.The obtained results showed that the fabricated sensor had a good selectivity for Trp against the other examined amino acids. Also, the potential applicability of the aptasensor was investigated by detecting the Trp in a complex media such as human blood plasma spiked with Trp.