Historical data from the Mussel Watch Programme (MWP) in South Africa from 1985 to

2008 were sampled during spring and autumn at selleck products spring low tide. Samples of M. galloprovincialis were collected and analysed for metals (μg/g dry weight) by the Department of Environmental Affairs and seven metals were analysed for this study (Cd, Cu, Pb, Zn, Hg, Fe and Zn). Prior to 1995, all MWP samples (n = 702, average mussel length = 60.8 mm) were collected and processed according to the methods used by Watling and Watling (1976). In brief, soft tissue of M. galloprovincialis were weighed and then dried at 105 °C for 48 h. The tissue was then digested with redistilled, analar-grade nitric acid and the solution was allowed to evaporate. The residue was redissolved GSI-IX in a 4:1 nitric-perchloric acid mixture and the solution dried at about 250 °C. This residue was then dissolved in 10 mL of 0.1 mol/L nitric acid. Metal concentrations in solution were determined by Atomic Absorption Spectrophotometry. The results were expressed as metal concentration in mussel tissue of whole organisms (μg/g dry tissue). Watling and Watling (1976) made no reference to any form of quality control and it is thus assumed that no certified reference material was used. After 1995, mussels (n = 802, average mussel length = 62.2 mm) were depurated in tanks filled with flowing

sea water for 24 h, whereafter they were freeze-dried for approximately 3 days and metal concentrations aminophylline determined as above. Quality control of metal concentrations was verified by including blanks and certified reference material (CRM) (DORM-2, dogfish muscle tissue, National Research Council Canada).

No data was available regarding recoveries for the entire period but data research reports at the Department indicated that recoveries were within 10% as the institution adheres to stringent quality assurance processes. All statistical data analysis was done using Statistica v10 (Stat. Soft. Inc.). The effects of time (annually and per season) and location on metal concentration in mussels were analysed and presented as mean concentrations (±SD) and further analysis using one-way ANOVA for single factors (year, season or site) and multiway ANOVA to test the effects of time (year and season) and location (distance from control site) on metals (Cd, Cu, Zn, Pb, Hg, Fe and Zn). Prior to the use of the parametric tests, the data were tested for normality and homogeneity of variances using Kolmogorov–Smirnov and Levene’s tests respectively (Townend, 2002). If the data did not meet the assumptions of the tests, the data were log10-transformed prior to analysis. For ANOVA analysis, post hoc Tukey tests were done. Error bars in graphs indicate standard error of the mean. Differences between seasonal metal concentrations were done using one way ANOVA and significant differences indicated at p < 0.05.

LA was efficient to restore the normal levels of PAP-SF and to decrease DPPIV-SF activity of envenomed mice to lower levels than the controls, whereas SA was efficient to restore the normal levels

of APN-SF. Both LA and SA were also able to mitigate the effect of the LD50 of vBj on APB activity, but they did not alter the effect of this dose of venom on the PIP-SF in the renal cortex. Table 4 also shows that the protein content of MF of the renal cortex decreased under the action of LD50 of vBj. The level of DPPIV-MF activity was not affected, but all other AP under study in the MF of the renal cortex were susceptible to this dose of vBj, that PLX-4720 price is: APA and CAP increased, and APN-MF, PIP-MF and PAP-MF decreased compared with controls. SA abolished the effect of LD50 of vBj on APN-MF and both SA and LA were able to mitigate the

effect of this dose of vBj on the levels of APA-MF and CAP-MF in the renal cortex. However, both drugs were unable to alter the effects of LD50 of vBj on the protein content of MF and on the activity levels of PIP-MF and PAP-MF in the renal cortex of envenomed mice. Furthermore, the association of these drugs with the LD50 of vBj promoted a significant decrease in DPPIV-MF activity in the renal cortex. Table 5 shows that the protein content in the SF of the renal medulla was not affected by the LD50 of ABT 199 vBj, as occurred in this same fraction of the renal cortex. However, similarly to the pattern that occurred in the SF of the renal cortex, all AP activities under study in the SF of the renal medulla were susceptible to this dose of vBj, that is: APB and DPPIV-SF increased, and APN-SF, PIP-SF and PAP-SF decreased in relation to the controls. LA was efficient to mitigate the

effects of vBj on the activities of Dichloromethane dehalogenase APB and PAP-SF. LA and SA were efficient to restore the normal levels of DPPIV-SF, but they did not alter the effect of the LD50 of vBj on APN-SF and PIP-SF activities in the renal medulla. Table 5 also shows that the protein content in the MF of the renal medulla decreased under the action of the LD50 of vBj, as occurred in the MF of the renal cortex ( Table 4). In the renal medulla, the levels of APN-MF and DPPIV-MF activities were not significantly affected, but all other AP activities under study in the MF of the renal medulla were susceptible to this dose of vBj, that is: APA increased, and PIP-MF, CAP and PAP-MF decreased in relation to the controls. Both drugs, LA and SA, were efficient only for restoring the normal levels of APA, but they did not alter the effects of the LD50 of vBj on the protein content in the MF and on the activities of PIP-MF, CAP and PAP-MF in the renal medulla. Both drugs also decreased the activity of DPPIV in the MF of the renal medulla when associated with the LD50 of vBj, as occurred in the MF of the renal cortex.

After

EUS TCB diagnosis of type 2 AIP, prednisone therapy was administered in 4 patients (patients 1, 2, 6, and 7), resulting in complete clinical resolution without recurrence over a mean follow-up of 22 months (range, 1.5-54 months). Three patients (patients 4, 8, and 9) with AIP were not given steroids due to insufficient disease severity. Of these, 1 patient (patient 4) remains under close supervision at our facility, whereas the other 2 are being followed by their referring institution. The use of TCB in the pediatric patient population has been limited PD0332991 price by a difficult-to-use TCB needle, smaller pancreas size in children, relative paucity of indications, uncertain role of TCB, and heightened concern regarding its safety in pediatric patients. Our data, however, suggest the potential utility and safety of EUS TCB in a pediatric population. The diagnostic yield of EUS TCB in our patient population was 86%, which is comparable with that reported in adults.2, 9, 10 and 11 Only 1 Selleck 3MA patient with AIP had nondiagnostic pathology, but the EUS imaging features suggested the diagnosis of AIP, which had not been previously suspected. The EUS TCB diagnosis of AIP significantly altered the management of our patients, leading directly to steroid therapy in most. For the 3 patients with AIP and mild symptoms,

the ability to obtain a definitive diagnosis now allows careful monitoring and disease-specific therapy if subsequently clinically required. In these patients, we opted to obtain TCB specimens to optimize the chance for diagnosis given the lower diagnostic sensitivity of EUS TCB when obtained after steroid therapy or during later disease stages. Early diagnosis of pancreatic pathology, especially AIP,

allows for timely and disease-specific therapy Sorafenib datasheet and may help prevent disease progression to advanced usual chronic pancreatitis. Histologic confirmation also avoids the risks associated with indiscriminate steroid therapy for incorrectly presumed AIP. The diagnosis of AIP, although always challenging, is particularly difficult among pediatric patients given their tendency for type 2 disease. Type 2 AIP was the most common diagnosis in this patient population, as opposed to type 1 AIP, which accounts for 80% of AIP in the general U.S. population.12 Type 2 AIP is typically found in younger patients than type 1 AIP, but AIP in general is thought to be uncommon in the pediatric population.13 Diagnosis of type 2 AIP requires histologic confirmation, which was achieved in 86% of our pediatric patients using EUS TCB.14 Our experience also suggests the potential under-recognition of this disease, especially in the pediatric population, may be in part due to the reluctance to obtain histologic verification. However, our study suggests that EUS TCB may be a safe and feasible diagnostic tool for AIP in children in the appropriate clinical setting.

We calculated height-for-age Z-score according to the United States Centers for Disease Control standards of recumbent length Z-scores, birth to 24 months, and stature

Z-scores, 2–20 years in centimeters, by gender and age. 10 Fourteen eGFR equations were included and their respective values for 81 patients were compared against the mGFRs. This retrospective study was approved by the Lurie Children’s Hospital of Chicago Institutional Review Board. We measured iohexol in www.selleckchem.com/products/Vincristine-Sulfate.html serum by a validated liquid chromatography tandem mass spectroscopy method from 4 serial blood samples collected at 10, 30, 120, and 300 minutes postiohexol injection with the clearance calculated using the concentration of iohexol as a function of time in 2 curves (fast and slow plasma disappearance).9 Scr was measured using an isotope-dilution mass spectrometry (IDMS)-traceable enzymatic method on the Roche Cobas

6000, following the Food and Drug Administration cleared procedure for Roche or Hitachi Cobas C systems. Blood urea nitrogen and cystatin C were analyzed in serum on the Roche Cobas 6000, following the Food and Drug Administration cleared procedures for Roche or Hitachi Cobas OSI-744 order C systems. The cystatin C method on the Roche Cobas 6000 uses an automated particle-enhanced immunoturbidimetric assay (PETIA). A total of 14 eGFR equations were selected to calculate eGFR (Table I). These include 5 equations based on Scr alone, 5 based on Scys alone, and 4 based on combinations of both. The method of testing Scys was particle-enhanced nephelometric immunoassay (PENIA) in Filler et al,16 Bouvet et al,17

Chehade et al,18 and Schwartz et al4 and 11 equations. The others used the PETIA method. The method of testing Scr was Jaffe method in Gao et al,12 Bouvet et al,17 and Chehade et al18 equations. The others used the enzymatic assay. Continuous data were described as the mean ± standard deviation, median, and interquartile range (IQR), and categorical variables were expressed as cases or percentages. Differences between eGFR and mGFR were analyzed by the nonparametric Wilcoxon test, because the data were not normally MRIP distributed. Correlations between eGFR and mGFR were established based on the Spearman correlation. Bland-Altman analysis was used to compare eGFR with mGFR using the average of the overall mean ± standard deviation and the precision was represented as the width between the 95% limits of agreement, wherein the smaller the limits of agreement, the greater the precision. Regression analysis and scatterplot analysis were used to compare the agreement between eGFR and mGFR. Three parameters used to assess the performance of eGFR equations relative to mGFR were as follows: • Bias (median difference between mGFR and eGFR) and absolute bias (median difference in |mGFR − eGFR|; We selected P < 0.05 a priori to be statistically significant.

MAANOVA revealed a total of 814 genes that were differentially expressed (false discovery rate (FDR)-corrected p ≤ 0.05) in exposed groups relative to their time-matched controls in both lung and liver for at least one dose ( Supplementary Table 3). The complete microarray datasets are available through the Gene Expression Omnibus at NCBI (http://www.ncbi.nlm.nih.gov/geo/), accession number GSE24751. Of the 814 genes, 269 were statistically significant with fold changes greater than 1.5 in both lung DZNeP mouse and liver at the highest dose (300 mg/kg) and 87 in the lower dose group (150 mg/kg). A very large fold change was noted for two detoxification enzymes

cytochrome p450 1A1 (Cyp1A1; 130–160 fold) and flavin containing monooxygenase 3 (Fmo3; 55–160 fold) in liver. Similarly, in the lungs, phase 1 enzymes Cyp1A1 and Cyp1B1 genes were the top two genes on the list with 25–50 fold induction ( Supplementary Table 3). The liver had 1151 genes that were statistically significant with fold changes greater than 1.5 at the high dose and 390 at the low dose. Pathway analysis on the genes showing fold changes higher than 1.5 in 300 mg/kg dose group in both lung and liver tissues identified Dasatinib pathways involved in oxidative stress, xenobiotic metabolism signalling, AHR signalling, and

glutathione metabolism as the commonly altered pathways. The main differences between the two tissues included negative regulation of genes associated with B cell receptor signalling and primary immunodeficiency in lungs compared to the liver. Details of liver transcriptome analyses for all doses and time points will be published separately. Agilent arrays containing 567 mouse probes were used

to examine changes in miRNAs in the Resminostat lungs of mice following exposure to BaP. Overall, 13 miRNAs in the high dose group (300 mg/kg) and 9 miRNAs in the low dose group (150 mg/kg) were significantly differentially regulated with fold changes greater than 1.5 and FDR p ≤ 0.05. miRNAs miR-34c, miR-34b-5p, miR-29b, miR-141, miR-199a-5p, miR-125a-5p and miR-200c were upregulated in one or both of the dose groups ( Table 5). These miRNAs are reported to be implicated in growth suppression, cell cycle, apoptosis, and tumour suppressor activity. Downregulated miRNAs included miR-122, miR-142-3p, miR-144, and miR-142-5p, miR-150 and miR-451 ( Table 5), which are associated with tumour suppression, hematopoiesis, erythroid differentiation and immune response. Complete miRNA microarray data are available through the Gene Expression Omnibus at NCBI (http://www.ncbi.nlm.nih.gov/geo/), accession number GSE24751. Real-time RT-PCR analysis confirmed the altered expression of miR-122, miR-142-5p, miR-29b, miR-34c, miR-34b-5p and miR-150 ( Fig. 1). We used TargetScan mouse 5.1 (Friedman et al., 2009 and Lewis et al.

expasy.org/tools/). Results from the hemolytic assays were expressed as mean ± SEM (Standard Error of the Mean). They were evaluated using two-way analysis of variance (ANOVA) followed by the Bonferroni post hoc test. Differences were considered significant at *p < 0.05.

Isolation of the cytolysin of S. plumieri venom was achieved in three steps. The first step involved fractionation of the crude venom by ammonium sulfate precipitation. The cytolytic toxin in venom was precipitated in high yield (80%), by 35% of salt saturation and named cytolytic fraction Inhibitor Library cell assay I (CF-I, Table 1). The 15% ammonium sulfate precipitate fraction and final supernatants fluids after removing 35% precipitated proteins showed very low hemolytic activity (data no shown). CF-I was resolved into four major peaks using hydrophobic interaction

chromatography. Strong hemolysis activity was detected in the fractions associated with the peak eluted at (NH4)2SO4 concentration of approximately 0.2 M (Fig. 1A). This material was grouped and named CF-II (Table 1). Subsequent fractionation of CF-II by anion exchange chromatography (Fig. 1B) resulted in eluting the hemolytic fraction as the forth protein peak eluted this website at a NaCl concentration of approximately 0.4 M (Table 1). This material corresponded to Sp-CTx and it migrated as a 71 kDa band upon SDS-PAGE (Fig. 1B, inset lane B), under reducing conditions. A quantitative evaluation of the hemolytic activity showed an EC50 of 282 ng/mL for CF-I, 111 ng/mL for CF-II and 25 ng/mL for Sp-CTx, which were approximately 2, 5 and 24 fold more hemolytic than crude venom (EC50 = 592 ng/mL, Table 1), respectively. The purification scheme of Sp-CTx is summarized in Table 1. SDS-PAGE analyses of Sp-CTx, under reducing condition, revealed a band of approximately 71 kDa (Fig. 1, inset, lane B) whereas under non-reducing condition an additional diffuse band of approximately 150 kDa was also observed (Fig. 1, inset, lane C). Two-dimensional (2D) electrophoresis revealed that the isoelectric

point (pI) of Sp-CTx ranges from 5.8 to 6.4 (data not shown). The chemical cross-linking studies Casein kinase 1 demonstrated proteins bands at ≈150 and 280 kDa even at a low BS3 concentration (1 mM). Those bands are indicative of dimer and tetrameric aggregation (Fig. 2). Besides, the 71 kDa band was not observed in the presence of BS3. Efforts to determine the N-terminal sequence of Sp-CTx were unsuccessful. No sequencing signal was obtained even with considerable amount (250 pmol) of the toxin. The resistance to Edman degradation chemistry suggests that the N-terminus of Sp-CTx is blocked. However, thirty-seven Sp-CTx internal amino acid sequences were obtained by Orbitrap-MS analyses, after proteolytic fragmentation with trypsin from both 71 and 150 kDa SDS-PAGE protein bands (under non-reduction conditions).

3c. Finally, the MODIS-A Local Area Coverage (LAC) data with 1 km nominal resolution are displayed in Fig. 3d. Note that the AMT data selleck screening library are not included in Fig. 3. The error statistics for data shown in Fig. 3 are summarized in Table 2. The categorization of data into 3 subsets (GAC, MLAC, LAC) does not show any evidence that either of the subsets has a much better statistics than the other data subsets. The R2 coefficient for all data subsets is about 0.8 if AMT data are not included. The lowest mean

absolute percentage error (MPE) of about 22% is for the MODIS-A LAC data set, while the lowest percentage of model bias (PBIAS) is for the SeaWiFS GAC data (about 1%). The results shown in Figure 2 and Figure 3 indicate that the performance of satellite POC algorithms is acceptable and comparable to the performance of the standard correlational satellite algorithms for chlorophyll (Chl) concentration (Bailey and Werdell, 2006). Similar conclusion has been reached by Duforet-Gaurier et al. (2010), but these authors used more limited data sets (27 data points). Allison Selleckchem Epacadostat et al. (2010) also concluded that the band ratio algorithm is currently the best option for estimating POC from ocean color remote sensing in the Southern Ocean, although they recommended a slightly modified version of the regional algorithm. In spite of

these results one has to recognize that the POC database (260 data points) is still modest when compared to global Chl matchup database (∼2500 data points in Siegel et al., 2013), and more efforts are needed to carry out global POC measurements to increase this database in the future. In addition, historically much less efforts have been devoted

to establishing robust POC in situ data Tolmetin collection protocols, and there have been no round robin or intercomparison experiments between different laboratories. More research efforts should be focused on this issue. In recent years, satellite-derived Chl data improved substantially our understanding of phytoplankton biomass and primary production distributions within the world’s oceans. However, of particular interest to ocean biogeochemistry and its role in climate change is not Chl, but carbon. It is therefore important to continue the experimental and conceptual work to improve the reliability of in situ and satellite POC determinations. Another challenging task for the ocean color methods is development of the capability to partition the POC stock into the living and non-living components (Behrenfeld et al., 2005). In our final word we would like to stress that even if scientists continue to strive to decrease errors and improve satellite methods, the substantial scientific benefits from use of large scale ocean color satellite observations are unquestionable. None declared. The authors would like to thank all the people who were involved in the programs providing free access to the data sets used in this study. The historical field data were obtained from the U.S.

At equal temperature, a reduction in CO2 partial pressure can lead to precipitation of calcite. These effects are also observed during lab experiments with 14 Dutch groundwater samples (Willemsen and Appelo, 1985). Finally, the transport of contaminants to the deeper groundwater can be accelerated by mixing processes. For contaminants wherefore the

degradation depends on redox conditions, mixing can create either more or less favorable conditions for degradation (van Oostrom et al., 2010 and Zuurbier et al., 2013). The extent to which ATES Cyclopamine solubility dmso systems mix different groundwater types depends on the screen length, sealing practice and water quality distribution in the aquifer surrounding the well screens. To achieve the desired flow rates, water is often extracted over a large portion of the aquifer. Where water quality

differences are present over the screen length, mixing may lead to changes in groundwater composition around the ATES wells. The effects of mixing in the extraction wells are comparable to the effects observed in drinking water wells, including clogging. However, the effects in the extraction wells of an ATES system are expected to be smaller since drinking water wells only produce groundwater Fulvestrant and, therefore continously pull water quality transition zones toward the wells. ATES wells on the other hand usually switch pumping direction twice a year, so that the main share of the water that is pumped has already been pumped and mixed in the previous season. As a consequence drinking water wells have a much higher probability to mix different types of groundwater. In ATES systems on the other hand, the pumped, mixed groundwater is re-injected into the aquifer in a nearby well, which is (usually) not the case for drinking water wells. Geochemical changes related to the

injection of water are discussed in the context of Aquifer Storage and Recovery (ASR) (Descourvières et al., 2010, Prommer and Stuyfzand, 2005 and Pyne, 2005). In ASR systems, often oxic (surface) Cyclin-dependent kinase 3 water is injected into anoxic aquifers and the geochemical effects are therefore larger than for ATES systems in which (mixed) water from the same aquifer is re-injected. In the storage volume, the native water is replaced by the injected water, and a new hydrochemical en geochemical equilibrium will be installed over time. A field and modeling study in the Netherlands (Bonte et al., 2013c) showed that ATES operation results in homogenization of the natural redox zoning in the aquifer, which may trigger secondary reactions such as mobilization of trace elements and organic carbon. However, the results of the investigated site showed that the observed concentration changes are sufficiently small to keep groundwater suitable for drinking water production from a chemical point of view.

The increasing intensity of the irrigation water diversion can also be seen from the dotted lines in Fig. 13. Water consumption increased more and more after 1980. Increase in the water demand and consumption by the industrial and service sectors is another reason for streamflow reduction in the Heihe River although they account for only around 10% of the total water use. China’s GKT137831 ic50 national economic reform began in 1978, and new industrial sectors have grown greatly since then, which drive up the GDP

rapidly. The GDP of Zhangye (Fig. 14(a)) increased notably after 1985, reached 21.2 billion Yuan in 2010 (about 600 folds of the 1950). The industry gross output (Fig. 14(b)) also increased notably after 1985, reached 9.84 billion Yuan

in 2010. According to the government statistical information, the proportion of outputs from the agriculture, industry and services, respectively, was 72%, 7%, and 21% in 1952, 47%, 30%, and 23% in 1980, and 28%, 36%, and 36% in 2012. The rapid rise in industry and services sectors led to a tremendous increase in water demand (Wang et al., 2009). Water consumption in the middle HRB increased from 5.13 × 108 to 8.71 × 108 m3 per year from 1985 to 2001 (Qi and Luo, 2005). The rapid development of agriculture and growth of economy began in the early 1980s. The downward abrupt change in the streamflow of the Zhengyixia station started in 1979, and significant upward abrupt change of the streamflow difference between Yingluoxia and Zhengyixia started

in 1982. The consistency in the timing confirmed that the decrease in the streamflow of the Heihe River were mainly due to local agricultural and AZD2281 economic development. For the middle and lower HRB where streamflow was greatly affected by human activities, the policy preference of the government was an important factor directly or indirectly contributing to streamflow changes. In the 1980s, the government desired to make the “Hexi Corridor” an important grain production base. The PLEKHM2 emphasis on grain production promoted the rapid advance of farming and irrigation projects. Unconstrained development resulted in streamflow being dried up in the lower HRB. The shortage of water for the lower HRB left people and the ecosystem in the downstream Gobi desert region to compete for limited water resources for survival. As a consequence, the fragile ecological system has been seriously damaged, and the conflict of water between the midstream and downstream became rampant. To restore the severely degraded ecosystem in the lower HRB, the government relied on water transfer projects to cope with water shortage. The central government had invested 2.3 billion Yuan to implement the EWDP in 2000. The increase in the streamflow to the downstream of HRB is a direct outcome of the EWDP. From 2000 to 2005, there had been 16 times of intermittent watering to the lower Heihe River with the total volume of 5.28 billion cubic meters (Guo et al., 2009).

83 mg/kg dose of LPS. Lower doses of the PRR agonists were administered in the LabMaster system because pilot experiments had shown that mice kept singly in the LabMaster system were more www.selleckchem.com/Androgen-Receptor.html sensitive to the treatments than group-housed animals. Mice receiving just one of the compounds (MDP, FK565 or LPS) were first injected with saline followed by the respective compound 4 h later. The first injection was given 3 h after start of the light phase. In experiments involving combination treatments, MDP + LPS and FK565 + LPS were given with a time

lag of 4 h between injection of the NOD and TLR agonist, since this timing has been shown to have the strongest priming effect on the immune system (Takada et al., 1990 and Takada and Galanos, 1987). Sickness responses were examined 3–4 h after injection and depression-like behavior 21–26 h post-treatment. The time points for the recording of the sickness responses were chosen according to the known time course of the sickness response to MDP or LPS (Frenois et al., 2007 and Engeland

et al., 2003). Sickness behavior has been shown to pass into depression-like behavior 1 day after injection of LPS (0.83 mg/kg), which find more was the reason for choosing the second time point (Frenois et al., 2007). Since MDP or FK565 alone did not induce behavioral changes in experiment 2.1 and only induced a modest cytokine response 3 h post-treatment, the single treatment with MDP or KF565 was not investigated in experiments 3.1 and 3.2. Mice were deeply anesthetized with Bumetanide pentobarbital (150 mg/kg i.p.). Blood was sampled by cardiac puncture using citrate (3.8%) as an anticoagulant. Following centrifugation for 10 min at 4 °C and 1000×g, blood plasma was collected and stored at −70 °C until assay. The plasma levels of corticosterone were determined with an enzyme immunoassay kit (Assay Designs, Ann Arbor, Michigan,

USA). According to the manufacturer’s specifications, the sensitivity of the assay is 27 pg/mL, and the intra- and inter-assay coefficient of variation amounts to 7.7% and 9.7%, respectively. Kynurenine and tryptophan were measured in plasma samples by high-performance liquid chromatography (HPLC) with ultraviolet detection (Herve et al., 1996). In brief, 100 μL plasma samples were deproteinized by adding of 100 μL of 5% (v/v) perchloric acid. After vortexing and 5 min centrifugation at 11,000×g, 20 μL of the clear supernatant was injected in the chromatographic system. Separations were achieved on a Chromolith RP18e column (100 × 4.6 mm, 5 μm, Merck Darmstadt, Germany) at 30 °C by isocratic elution with a mobile phase consisting of 50 mmol/L ammonium acetate, 250 mol/L zinc acetate and 3% (v/v) acetonitrile (pH 4.9) at a flow rate of 0.8 mL/min. Kynurenine and tryptophan were detected on a LaChrom UV-Detector Merck HITACHI L-7400 at 235 nm.