Increased Levels Of Tryptophan Research Articles

Cilia-deficient renal tubule cells are primed for injury with mitochondrial defects and aberrant tryptophan metabolism.

The exocyst and Ift88 are necessary for primary ciliogenesis. Overexpression of Exoc5 (OE), a central exocyst component, resulted in longer cilia and enhanced injury recovery. Mitochondria are involved in acute kidney injury (AKI). To investigate cilia and mitochondria, basal respiration and mitochondrial maximal and spare respiratory capacity were measured in Exoc5 OE, Exoc5 knockdown (KD), Exoc5 ciliary targeting sequence mutant (CTS-mut), control Madin-Darby canine kidney (MDCK), Ift88 knockout (KO), and Ift88 rescue cells. In Exoc5 KD, Exoc5 CTS-mut, and Ift88 KO cells, these parameters were decreased. In Exoc5 OE and Ift88 rescue cells they were increased. Reactive oxygen species were higher in Exoc5 KD, Exoc5 CTS-mut, and Ift88 KO cells compared with Exoc5 OE, control, and Ift88 rescue cells. By electron microscopy, mitochondria appeared abnormal in Exoc5 KD, Exoc5 CTS-mut, and Ift88 KO cells. A metabolomics screen of control, Exoc5 KD, Exoc5 CTS-mut, Exoc5 OE, Ift88 KO, and Ift88 rescue cells showed a marked increase in tryptophan levels in Exoc5 CTS-mut (113-fold) and Exoc5 KD (58-fold) compared with control cells. A 21% increase was seen in Ift88 KO compared with rescue cells. In Exoc5 OE compared with control cells, tryptophan was decreased 59%. To determine the effects of ciliary loss on AKI, we generated proximal tubule-specific Exoc5 and Ift88 KO mice. These mice had loss of primary cilia, decreased mitochondrial ATP synthase, and increased tryptophan in proximal tubules with greater injury following ischemia-reperfusion. These data indicate that cilia-deficient renal tubule cells are primed for injury with mitochondrial defects in tryptophan metabolism.NEW & NOTEWORTHY Mitochondria are centrally involved in acute kidney injury (AKI). Here, we show that cilia-deficient renal tubule cells both in vitro in cell culture and in vivo in mice are primed for injury with mitochondrial defects and aberrant tryptophan metabolism. These data suggest therapeutic strategies such as enhancing ciliogenesis or improving mitochondrial function to protect patients at risk for AKI.

Investigations Towards Tryptophan Uptake and Transport Across an In Vitro Model of the Oral Mucosa Epithelium.

Tryptophan is an essential amino acid and plays an important role in several metabolic processes relevant for the human health. As the main metabolic pathway for tryptophan along the kynurenine axis is involved in inflammatory responses, changed metabolite levels can be used to monitor inflammatory diseases such as ulcerative colitis. As a progenitor of serotonin, altered tryptophan levels have been related to several neurogenerative diseases as well as depression or anxiety. While tryptophan concentrations are commonly evaluated in serum, a non-invasive detection approach using saliva might offer significant advantages, especially during long-term treatments of patients or elderly. In order to estimate whether active transport processes for tryptophan might contribute to a potential correlation between blood and saliva tryptophan concentrations, we investigated tryptophan's transport across an established oral mucosa in vitro model. Interestingly, treatment with tryptophan revealed a concentration dependent secretion of tryptophan and the presence of a saturable transporter while transport studies with deuterated tryptophan displayed increased permeability from the saliva to the blood compartment. Protein analysis demonstrated a distinct expression of L-type amino acid transporter 1 (LAT1), the major transporter for tryptophan, and exposure to inhibitors (2 -amino-2-norbornanecarboxylic acid (BCH), L-leucine) led to increased tryptophan levels on the saliva side. Additionally, exposure to tryptophan in equilibrium studies resulted in a regulation of LAT1 at the mRNA level. The data collected in this study suggest the participation of active transport mechanisms for tryptophan across the oral mucosa epithelium. Future studies should investigate the transport of tryptophan across salivary gland epithelia in order to enable a comprehensive understanding of tryptophan exchange at the blood-saliva barrier.

Increased dietary 5-hydroxytryptophan reduces fearfulness in red junglefowl hens (Gallus gallus).

Our production animals typically suffer poor welfare, which can be revealed by measuring the affective state these animals are in. Negative affective state is linked to poorer welfare, and can be measured as fearfulness. While continuing to research how to improve animal welfare, a compliment to reduce negative affective state could therefore be to reduce individuals' fearfulness, similar to how negative affective states are medicated in humans. A proposed mechanism for this is via the monoaminergic systems. This is based on previous studies across species that have linked the serotonergic system and fear-related behaviour. We here aimed to experimentally manipulate the serotonergic system in red junglefowl hens (Gallus gallus), the main ancestor of all domesticated chickens. We measured fearfulness as latency remaining immobile in a tonic immobility test, and did so both before and after our experimental manipulation. We set out to experimentally manipulate the serotonergic system via sub-chronic dietary treatment of 5-hydroxytryptophan (the precursor to serotonin). Our dietary manipulation of 5-hydroxytryptophan significantly reduced measured fearfulness in the manipulated hens, while latency in tonic immobility did not significantly change in our unmanipulated, control hens. This finding is promising since it indicates that increased tryptophan levels can be used to reduce fearfulness. Additionally, our result suggests that this can be done non-invasively via food (instead of injections), thus presenting a potentially feasible manipulation also for larger settings. Nevertheless, the serotonergic system is complex and its role in modulating behaviour in the fowl should be explored further to evaluate our findings, and more directly explored also in a production setting.

Kynurenine Pathway Regulation at Its Critical Junctions with Fluctuation of Tryptophan.

The kynurenine pathway (KP) is the primary route for the catabolism of the essential amino acid tryptophan. The central KP metabolites are neurologically active molecules or biosynthetic precursors to critical molecules, such as NAD+. Within this pathway are three enzymes of interest, HAO, ACMSD, and AMSDH, whose substrates and/or products can spontaneously cyclize to form side products such as quinolinic acid (QA or QUIN) and picolinic acid. Due to their unstable nature for spontaneous autocyclization, it might be expected that the levels of these side products would be dependent on tryptophan intake; however, this is not the case in healthy individuals. On top of that, the regulatory mechanisms of the KP remain unknown, even after a deeper understanding of the structure and mechanism of the enzymes that handle these unstable KP metabolic intermediates. Thus, the question arises, how do these enzymes compete with the autocyclization of their substrates, especially amidst increased tryptophan levels? Here, we propose the formation of a transient enzyme complex as a regulatory mechanism for metabolite distribution between enzymatic and non-enzymatic routes during periods of increased metabolic intake. Amid high levels of tryptophan, HAO, ACMSD, and AMSDH may bind together, forming a tunnel to shuttle the metabolites through each enzyme, consequently regulating the autocyclization of their products. Though further research is required to establish the formation of transient complexation as a solution to the regulatory mysteries of the KP, our docking model studies support this new hypothesis.

Untargeted Metabolomics Reveals Metabolic Stress Alleviation by Prepartum Exercise in Transition Dairy Cows.

Prepartum exercise (PA) has been proposed as a strategy for the peripartum management of dairy cows; however, the mechanism by which PA affects metabolism has not been elucidated. Here, we investigated the metabolic changes in transition dairy cows with PA. Holstein transition multiparous dairy cows were assigned to an exercise (n = 12) or a control (n = 12) group; the cows in the exercise group walked for a targeted 45 min at 3.25 km/h, two times a day. Plasma non-esterified fatty acid (NEFA), β-hydroxybutyric acid (BHBA), glucose, and triglyceride levels were measured, and metabolic profiles were analyzed using untargeted mass spectrometry. Compared with those in the control group, the concentrations of NEFA at −7 d, glucose at 0 d, and BHBA at +7 d relative to calving were considerably decreased in the exercise group. Untargeted metabolomics analysis revealed differences in the levels of key metabolites, including kynurenine, tryptophan, homovanillic acid, dopamine, cis-9-palmitoleic acid, and palmitic acid, between the exercise and control group cows. This study suggests that PA may decrease homovanillic acid and cis-9-palmitoleic acid levels and increase tryptophan levels to alleviate the metabolic stress in dairy cows during calving, thereby improving postpartum health.

Resistance Exercise Modulates Kynurenine Pathway in Pancreatic Cancer Patients.

The aim of this study was to investigate the impact of Supervised and Home-based resistance exercise on the Kynurenine pathway in patients with pancreatic cancer who underwent surgery and chemotherapy. In the SUPPORT study, adult pancreatic cancer patients were randomized to intervention programs of 6-month (1) a Supervised moderate-to-high-intensity progressive resistance training or (2) unsupervised Home-based resistance training, or (3) to a standard care patient Control group. Serum levels of kynurenine, tryptophan and IL-6 were assessed for 32 participants before, after 3 months and after 6 months of exercise intervention. Group differences were investigated using analysis-of-covariance. Patients in the Supervised training group showed decreased levels of serum kynurenine and kynurenine/tryptophan ratio (p = 0.07; p = 0.01 respectively) as well as increased Tryptophan levels (p = 0.05) in comparison to Home-based and Control group over time. The Home-based exercise group had significant increased kynurenine and kynurenine/tryptophan ratio levels. IL-6 levels decreased over the first three months for both intervention groups as well as the Control group (Supervised: p < 0.01, Home-based: p < 0.010, Control group: p < 0.01). Supervised resistance exercise might positively regulate the Kynurenine pathway and downregulate the kynurenine/tryptophan (indicative of IDO/TDO enzyme) levels, hence modulating the immune system.

COVID-19, interferons, and depression: A commentary

An option currently being explored for the treatment of COVID-19 is the use of interferons (INFs), either alone or in combination with other antiviral agents. INFs are known to shift the metabolism of tryptophan (TRP) away from its role as a precursor of serotonin. For some patients, reduction in TRP levels may either expose an underlying vulnerability to depression or trigger a de novo episode of depression. This Commentary discusses the pathway involved and recommends in-hospital augmentation with foods or supplements that increase TRP levels for COVID-19 patients treated with INFs. Selective serotonin reuptake inhibitors may also be tried if the depressive symptomatology is not short-lived.

Tryptophan levels associate with incident cardiovascular disease in chronic kidney disease.

BackgroundNon-traditional risk factors like inflammation and oxidative stress play an essential role in the increased cardiovascular disease (CVD) risk prevalent in chronic kidney disease (CKD). Tryptophan catabolism by the kynurenine pathway (KP) is linked to systemic inflammation and CVD in the general and dialysis population. However, the relationship of KP to incident CVD in the CKD population is unknown.MethodsWe measured tryptophan metabolites using targeted mass spectrometry in 92 patients with a history of CVD (old CVD); 46 patients with no history of CVD and new CVD during follow-up (no CVD); and 46 patients with no CVD history who developed CVD in the median follow-up period of 2 years (incident CVD).ResultsThe three groups are well-matched in age, gender, race, diabetes status and CKD stage, and only differed in total cholesterol and proteinuria. Tryptophan and kynurenine levels significantly decreased in patients with ‘Incident CVD’ compared with the no CVD or old CVD groups (P = 5.2E–7; P = 0.003 respectively). Kynurenic acid, 3-hydroxykynurenine and kynurenine are all increased with worsening CKD stage (P < 0.05). An increase in tryptophan levels at baseline was associated with 0.32-fold lower odds of incident CVD (P = 0.000014) compared with the no CVD group even after adjustment for classic CVD risk factors. Addition of tryptophan and kynurenine levels to the receiver operating curve constructed from discriminant analysis predicting incident CVD using baseline clinical variables increased the area under the curve from 0.76 to 0.82 (P = 0.04).ConclusionsIn summary, our study demonstrates that low tryptophan levels are associated with incident CVD in CKD.

Abstract TP104: Post Stroke Depressive-Like Behaviors in Middle-Aged Female Rats is Associated With Gut Dysbiosis and Improved by Mir363-3p

Background: Post-stroke depression (PSD) occurs in one-third of stroke survivors, is more prevalent in women, and negatively impacts quality of life and recovery. Our previous work showed that acyclic middle-aged female rats display depressive behaviors after stroke, and IV treatment with mir363-3p attenuates these behaviors. Serotonin has been implicated in affective behaviors, and loss of this neurotransmitter is also associated with decreased neurogenesis in the hippocampus. The present study determined (a) whether stroke affected tryptophan, a gut metabolite, and precursor for serotonin, and cell proliferation in the hippocampus and (b) whether serum tryptophan levels and hippocampal cell proliferation was restored by mir363-3p. Methods: Ischemic stroke was induced by stereotaxic delivery of endothelin-1 to the MCA of middle-aged female rats. Animals received a single injection of mir363-3p or scrambled oligos 4h after stroke. Depressive-like behaviors were assessed by the Effort-based sucrose consumption test, Social Interaction and Forced Swim Test 3 months after stroke. Blood was collected at termination. Serum tryptophan levels were quantified by ELISA. Bacterial composition was analyzed by 16S sequencing of fecal samples. Ki67 immunohistochemistry was performed on brain tissue collected at termination and quantified within the dentate gyrus by two blind observers. Results: Animals subjected to stroke exhibited depressive-like behaviors and had decreased tryptophan levels as compared to sham treated animals. This is also accompanied by microbiota dysbiosis as measured by an elevated ratio of Firmicutes to Bacteroidetes. Mir363-3p treatment increased tryptophan levels as compared to scrambled controls and were no different from sham (non-stroke) animals. Moreover, miR363-3p treated rats exhibit a significantly more dentate gyrus-specific Ki67 expression a proliferation marker. Conclusions: Together, these data indicate that miR363-3p attenuates post stroke depression via a mechanism that prevents microbiota dysbiosis and increases hippocampal cell proliferation.

Bioactive Amines Screening in Four Genotypes of Thermally Processed Cauliflower.

Biogenic amines are important indicators of food quality with recognized antioxidant capacity. Diets that are rich in these compounds promote several benefits for human health, although the consumption in excess may result in food poisoning. This study aims to screen the levels of biogenic amines in four colored cauliflowers, before and after cooking (boiling, steaming, and microwaving). In addition, the levels of tryptophan and 5-hydroxytryptophan, two serotonin precursors, were analyzed. Our results reveal that thermal processing shows a tendency to increase tryptophan levels and reduce 5-hydroxytryptophan in colored cauliflowers. A reduction of the tryptophan and increase in serotonin contents in ‘Cheddar’, steamed or microwaved, was observed. A higher level of histamine was observed in the genotype ‘Forata’ after cooking, whereas melatonin levels were higher after steaming and microwaving. The lowest levels of biogenic amines and amino acids were observed in ‘Graffiti’. All the colored cauliflowers that were analyzed presented a chemical quality index (CQI) below the pre-established limits, indicating that are safe for consumption, even after cooking. We conclude that the levels of biogenic amines and amino acids in colored cauliflower are safe for human consumption and do not present health risks. Therefore, the consumption of these genotypes, raw or cooked, is a good source of bioactive compounds.

Integrated constraints based analysis of an engineered violacein pathway in Escherichia coli

Strategies towards optimal violacein biosynthesis, a potential drug molecule, need systems level coordination of enzymatic activities of individual genes in a multigene operon vioABCDE. Constraints-based flux balance analysis of an extended iAF1260 model (iAF1260vio) with a reconstructed violacein module predicted growth and violacein yields in Escherichia coli accurately. Shadow price (SP) analysis identified tryptophan metabolism and NADPH as limiting. Increased tryptophan levels in Δpgi & ΔpheA were validated using in silico gene deletion analysis. Phenotypic phase plane (PhPP) analysis highlighted sensitivity between tryptophan and NADPH for violacein synthesis at molar growth yields. A synthetic VioABCDE operon (SYNO) sequence was designed to maximize Codon Adaptive Index (CAI: 0.9) and tune translation initiation rates (TIR: 2–50 fold higher) in E. coli. All pSYN E. coli transformants produced higher violacein, with a maximum six-fold increase in yields. The rational design E. coli: ΔpheA SYN: gave the highest violacein titers (33.8 mg/l). Such integrated approaches targeting multiple molecular hierarchies in the cell can be extended further to increase violacein yields.

INFLUENCE OF DITARY TRYPTOPHAN SUPPLEMENTAL ON PRODUCTIVE PERFORMANCE OF SINAI BEDOUIN LAYER HENS

This experiment was conducted in the Poultry Research Farm and the Poultry Nutrition Laboratory, Faculty of Agriculture, Minufia University, Shebin El-kom, in order to investigate the effect of dietary tryptophan (Trp) supplementation on Production performance, egg quality traits and some blood constituents of Sinai Bedouin layer hens were fed on supplementation during the period of 26 to 38 wks of age were determined. Economic efficiency and phonetic correlation between some productive parameters at the different levels of tryptophan supplementation were calculated. Results showed that final body weight and weight gain were significantly (P≤ 0.05) higher by increasing of tryptophan level. Hen–day production percentage, egg number, egg weight and egg mass were significantly improved as the level of dietary tryptophan increased. Feed conversion ratio was significantly (P ≤ 0.05) improved. Pronounced effects on egg shell percentage and thickness were significantly noted. Albumen, yolk and Haugh units were improved by adding tryptophan at the level of 0.23% of laying diets. Other traits of egg quality were not significantly affected by higher tryptophan levels. High positive phenotypic correlations between some productive parameters and final body weight was noticed by increasing tryptophan levels. Significant phenotypic correlation between egg weights and some egg quality traits by increasing tryptophan levels was recorded. There was a significant increase in plasma total protein, globulin concentration resulted from dietary tryptophan. No significant differences were determined in transaminase enzymes (ALT and AST) plasma blood concentration between all treatments significantly reduced the levels of total cholesterol and triglycerides concentrations. Finally, it could be concluded that dietary tryptophan might have positive effects on productive performance, economic efficiency and health status of layer hens. In respect of Sinai Bedouin laying hens diets 0.23% tryptophan could be recommended for improve most of economic traits.

Bile Acids and Tryptophan Metabolism Are Novel Pathways Involved in Metabolic Abnormalities in BPA-Exposed Pregnant Mice and Male Offspring.

Increasing evidence has demonstrated that exposure to endocrine-disrupting chemicals impacts maternal and fetal health, but the underlying mechanisms are still unclear. We previously showed that dietary exposure to 10 µg/kg body weight (bw)/d and 10 mg/kg bw/d of bisphenol A (BPA) during pregnancy induced metabolic abnormalities in F1 male offspring and gestational glucose intolerance in F0 pregnant mice. The aim of this study was to elucidate the underlying etiologies of BPA exposure-induced metabolic disease by analyzing the male fetal liver metabolome. Using the Metabolon Discover HD4 Platform, our laboratory identified metabolic pathways that were altered by BPA exposure, including biochemicals in lipid and amino acid metabolism. Specifically, primary and secondary bile acids were increased in liver from BPA-exposed embryonic day 18.5 male fetuses. We subsequently showed that increased bile acid was associated with a defective farnesoid X receptor-dependent negative feedback mechanism in BPA-exposed fetuses. In addition, through metabolomics, we observed that BPA-exposed fetuses had elevated tryptophan levels. Independent liquid chromatography and mass spectrometry measurement revealed that BPA-exposed dams also had increased tryptophan levels relative to those of controls. Because several key enzymes in tryptophan catabolism are vitamin B6 dependent and vitamin B6 deficiencies have been linked to gestational diabetes, we tested the impact of vitamin B6 supplementation and showed that it rescued gestational glucose intolerance in BPA-exposed pregnant mice. Our study has therefore identified two pathways (bile acid and tryptophan metabolism) that potentially underlie BPA-induced maternal and fetal metabolic disease.

Antidepressant-like effects of the xanthine oxidase enzyme inhibitor allopurinol in rats. A comparison with fluoxetine

Allopurinol is a xanthine oxidase enzyme inhibitor that is widely used for the treatment of hyperuricemia and gout. The activity of tryptophan 2,3-dioxygenase, which metabolizes tryptophan (TRP), is decreased by xanthine oxidase inhibitors, causing TRP levels in the body to be increased. Increases in TRP levels in the brain might have antidepressant effects. The purpose of this study is to evaluate the antidepressant effects of allopurinol compared to those of fluoxetine, which is a proven antidepressant. Thirty-two Wistar albino male rats were divided into four groups (control, 10mg/kg fluoxetine, 50mg/kg allopurinol, 50mg/kg allopurinol+10 mg/kg fluoxetine; n=8 per group), and forced swimming tests were performed before and after 14days of drug administration. Serotonin, 5-hydroxyindolacetic acid and uric acid levels were measured in blood samples after the final treatment. When allopurinol and fluoxetine were administered separately, a decrease in the duration of immobility and an increased duration of swimming were observed in the forced swimming test. The results showed similar antidepressant efficacies between allopurinol and fluoxetine. However, we found no statistically significant difference in the antidepressant effect of the combined therapy versus single drug therapy.

Effects of dietary l-tryptophan on the agonistic behavior, growth, and survival of freshwater crayfish Astacus leptodactylus Eschscholtz

In this study, the effects of dietary tryptophan (a precursor of 5-hydroxytryptamine, 5-HT, serotonin) on the agonistic behavior, growth, and survival of freshwater crayfish were investigated. For this aim, a control diet (D1) and three experimental diets (D2, D3, and D4) were prepared. D1 contained no additional tryptophan (TRP), but D2, D3, and D4 diets were supplemented with TRP at 0.50, 0.75, and 1.00 % in dry diet, respectively. The control contained 0.33 % TRP. Results revealed that higher supplemental dietary TRP levels caused a significant decrease in the aggressive behavior (P 0.05). In addition to these, supplemental dietary TRP gave rise to significantly higher specific growth rate (SGR) values in June and July (P < 0.05). For example, in June, it was 0.38 for the crayfish fed D4, 0.28 for the crayfish fed D3, 0.18 for the crayfish fed D2, and 0.13 for the crayfish fed control. However, in August, the highest SGR (0.56) was obtained from the crayfish fed control. The results also showed that the SGRs of females were lower than the males in June, July, and August (P < 0.05). In conclusion, this research shows that the aggressive behavior of A. leptodactylus can be suppressed, and rearing efficiency (i.e., increased growth and high survival rate) of this species can be improved by increasing TRP levels to 1.00 % in its diet.

B17 Delaying ageing and the ageing-associated decline in protein homeostasis by inhibition of tryptophan degradation

Toxicity of aggregation-prone proteins is thought to play an important role in ageing and age-related neurological diseases like Parkinson9s and Alzheimer9s disease. Here we identify tryptophan 2,3-dioxygenase (tdo-2), the first enzyme in the kynurenine pathway of tryptophan degradation, as a metabolic regulator of age-related α-synuclein toxicity in a C. elegans model. Depletion of tdo-2 also suppresses toxicity of other heterologous aggregation-prone proteins, including amyloid-β and polyglutamine proteins, and of endogenous meta-stable proteins that are sensors of normal protein homeostasis. This suggests that tdo-2 functions as a general regulator of protein homeostasis. Analysis of metabolite levels in C. elegans strains with mutations in enzymes that act downstream of tdo-2 indicates that this suppression of toxicity is independent of downstream metabolites in the kynurenine pathway. Depletion of tdo-2 increases tryptophan levels and feeding worms with extra L-tryptophan also suppresses toxicity, suggesting that tdo-2 regulates proteotoxicity through tryptophan. Depletion of tdo-2 extends lifespan in worms. Together these results implicate tdo-2 as a metabolic switch of age-related protein homeostasis and lifespan. With TDO and Indoleamine 2,3-dioxygenase as evolutionarily conserved human orthologs of TDO-2, intervening with tryptophan metabolism may offer avenues to reducing proteotoxicity in ageing and age-related diseases.

Possible Modulating Effects of Celecoxib (COX II Inhibitor) on Antide-pressant Action of Duloxetine (SNRI) in Stressed Mice

Rationale: In recent years, a potential link between inflammation and depression has been shown and the role of proinflammatory cytokines in the pathophysiology of major depressive disorders has been observed. Celecoxib (selective COX-II inhibitor) has been reported to inhibit the production of PGE-II and proinflammatory cytokines and also increase tryptophan levels and serotonin availability in depressed patients. On the other hand, duloxetine (potent SNRI) has been shown to be efficacious in inflammatory and acute pain models in rodents and synergistic interaction with NSAIDs. How- ever, the interactions of duloxetine with celecoxib are currently unknown. Objective: We evaluated the antidepressant effect of celecoxib (15, 30 mg/kg/day for 15 days, ip) alone and in combina- tion with duloxetine (5, 10 mg/kg/day for 15 days, ip) and also the biochemical parameters in stressed mice. Results: Pretreatment of celecoxib (15, 30 mg/kg) for 15 days to forced swim-induced stressed mice produced significant antidepressant effect which has been evidenced by decreased in immobility time in tail suspension test (TST). Celecoxib (30 mg/kg) also showed significant increase in locomotor activity and protective effect on biochemical parameters of oxi- dative stress by reversing stress-induced increase in TBARS and reduction in GSH levels. Pretreatment with combination of celecoxib with duloxetine (5, 10 mg/kg) showed significant antidepressant and neuroprotective effects against stress in- duced depression and oxidative damage in mice at both dose levels. Conclusions: This study demonstrated dose-dependent antidepressant action of celecoxib in stressed mice. The combina- tion of celecoxib with duloxetine further enhanced its antidepressant effect on TST in stressed mice. The treatment re- versed forced swim-induced elevation in TBARS levels and depleted glutathione activity, suggesting their antioxidant and protective role in brain.

Brain tryptophan rather than pH-value is altered as consequence of artificial postmortem interval and storage conditions

Brain bank centers around the world attempt to standardize postmortem brain collection and quality control. Antemortem as well postmortem factors may influence tissue quality. Previously, we could demonstrate that increased tryptophan (TRP) levels significantly correlate to prolonged postmortem interval (PMI) and storage duration, whereas pH-value altered merely as consequence of prolonged agonal state and ischemic brain damage additionally to repeated freeze and thaw cycles. Therefore, we aimed to investigate in artificial PMI conditions, with three brain tissue storage temperatures (4 °C, room temperature and 37 °C) as well as oxidizing conditions (open/close tube), whether TRP levels and pH-value alter. We could confirm that prolonged PMI at higher storage temperatures and oxidizing conditions significantly correlate to increased TRP levels, while pH-value did not correlate at all. In conclusion, from these results PMI intervals until autopsy should be kept as short as possible and storage until autopsy should be at 4 °C in order to preserve brain tissue quality as much as possible.

Kernel modifications and tryptophan content in QPM segregating generations

Maize has poor nutritional value due to deficiency of two essential amino acids - tryptophan and lysine. Although recessive opaque2 (o2) mutation significantly increases their content in the endosperm, incorporation of opaque2 into high yielding cultivars was not commercially successful, because of its numerous agronomic and processing problems due to soft endosperm. Quality protein maize - QPM has lately been introduced as opaque2 maize with improved endosperm hardness and improved agronomic traits, but mostly within tropical and subtropical germplasm. The ongoing breeding project at MRI includes improvement of MRI opaque2 lines and conversion of standard lines to QPM germplasm. The main selection steps in QPM breeding involve assessing kernel modifications and tryptophan level in each generation. Herein, we present the results of the analysis for these traits on F3 and BC1F1 generations of QPM x opaque2, opaque2 x QPM and standard lines x QPM crosses. The results showed that the majority the genotypes had kernel types 2 and 3 (good modifications). The whole grain tryptophan content in F3 and BC1F1 genotypes of crosses between QPM and opaque2 germplasm was at the quality protein level, with a few exceptions. All BC1F1 genotypes of standard lines x QPM had tryptophan content in the range of normal maize, while majority of F3 genotypes had tryptophan content at level of QPM. The progeny (with increased tryptophan levels) of QPM and opaque2 crosses had significantly higher tryptophan content compared to the progeny of crosses between standard and QPM lines - 0.098 to 0.114 and 0.080, respectively. All genotypes that had poorly modified kernels and/or low tryptophan content will be discarded from further breeding.

Shifts in the balance of brain tryptophan metabolism due to age and systemic administration of lipopolysaccharide

The kynurenine (KYN) pathway, which is initi-ated by indoleamine 2, 3-dioxygenase (IDO), is a key tryptophan (TRP) metabolic pathway. It shares TRP with the serotonin (5-HT) pathway. Because activation of the KYN pathway by proinflammatory cytokines induces depressive symptoms, shifts in the balance of TRP metabolism to the KYN pathway are closely related to the etiology of depression. In the present study, the influence of age on the effect of the inflammation response system (IRS) on brain TRP metabolism was investigated. Male ICR mice (PND21) were reared for 4 weeks (younger group) or until they reached 1 year of age (older group), and given an intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). The TRP, KYN, and 5-HT levels were measured in the prefrontal cortex, hippocampus, amygdala, and dorsal raphe nuclei. An increase in TRP and 5-HT levels was observed with age in all brain regions, whereas age was associated with decreases in KYN levels in the dorsal raphe nuclei. In all brain regions, LPS increased TRP levels, while it in-creased KYN levels in the prefrontal cortex and amygdala. Reduced KYN/5-HT ratios in all regions were observed with age, whereas increased KYN/5-HT ratios were observed with LPS in all regions except the dorsal raphe nuclei. Thus, age shifted the balance between the KYN and 5-HT pathways toward the 5-HT pathway, and countered the effects of LPS, which shifted the balance to the KYN pathway. These effects are relevant to the etiology of psychiatric disorders in elderly people.