Low Levels Of Amino Acids Research Articles

Modified throughput ninhydrin method for the qualitative assessment of dietary protein absorption in pig plasma.

Protein maldigestion and malabsorption lead to malnutrition and are a feature of exocrine pancreatic insufficiency (EPI). Although it is the current standard, measurement of nitrogen in stool to assess protease activity is indirect. Up to 80% of hydrolysed proteins appear in blood in the form of peptides, so we developed a method to measure peptide-derived amino acids in plasma as a relevant measure of proteolysis, verified its accuracy, precision, and linearity, and validated it in a porcine model. We modified a ninhydrin method. Large proteins were eliminated from plasma with 10 kDa-cut-off centrifugal filters. Free and total amino acids were measured in permeate before and after its hydrolysis. Peptide-derived amino acids were quantified by subtracting free amino acids from total amino acids. We verified the method in vitro and by comparing results in healthy and EPI pigs. The accuracy of the analysis was close to 100%, with excellent precision (mean relative standard deviation for low, medium, and high amino acid levels = 0.88%) and with stringent linearity (r2 = 0.986, %RE = 5.23). The high-throughput ninhydrin method detected levels of peptide-derived amino acids in vivo with maximal changes seen approximately 2 hours postprandially in young pigs. The AUC and Cmax were significantly higher in healthy compared to EPI pigs (P = .0026 and P = .0037, respectively). The high-throughput ninhydrin method is a sensitive, reliable, and practical method for the estimation of dietary peptide-derived amino acids. This assay endpoint could serve as a direct biomarker of protein digestion and absorption.

Comparison of gnotobiotic communities reveals milk-adapted metabolic functions and unexpected amino acid metabolism by the pre-weaning microbiome

ABSTRACT The intestinal microbiome during infancy and childhood has distinct metabolic functions and microbial composition compared to adults. We recently published a gnotobiotic mouse model of the pre-weaning microbiome (PedsCom), which retains a pre-weaning configuration during the transition from a milk-based diet to solid foods, leads to a stunted immune system, and increases susceptibility to enteric infection. Here, we compared the phylogenetic and metabolic relationships of the PedsCom consortium to two adult-derived gnotobiotic communities, Altered Schaedler Flora and Oligo-Mouse Microbiota 12 (Oligo-MM12). We find that PedsCom contains several unique functions relative to these adult-derived mouse consortia, including differences in carbohydrate and lipid metabolism genes. Notably, amino acid degradation metabolic modules are more prevalent among PedsCom isolates, which is in line with the ready availability of these nutrients in milk. Indeed, metabolomic analysis revealed significantly lower levels of total free amino acids and lower levels of specific amino acids abundant in milk (e.g. glutamine and glutamic acid) in the intestinal contents of adult PedsCom colonized mice compared to Oligo-MM12 controls. Metabolomic analysis of pre-weaning intestinal contents also showed lower levels of amino acids that are replete in milk compared to germ-free controls. Thus, enhanced amino acid metabolism is a prominent feature of the pre-weaning microbiome that may facilitate design of early-life microbiome interventions.

Dietary threonine influences antioxidant capacity and immune status in juvenile largemouth bass (Micropterus salmoides)

As a restricted amino acid, threonine (Thr) deficiency reduces antioxidant and immune capacity in fish. In the present study, six diets containing different Thr levels (1.39 %, 1.71 %, 1.95 %, 2.28 %, 2.58 %, and 2.65 %) were fed to largemouth bass (15±0.1 g) for 60 days. The present results showed that neither growth nor whole-body composition was affected by different Thr levels. And low dietary Thr levels (1.39 %) significantly decreased serum total protein (TP) and albumin (ALB) levels, but increased alkaline phosphatase (ALP) levels. And 1.39 % Thr diet significantly decreased liver catalase (CAT) activity but increased malonaldehyde (MDA) levels, and the maximum total antioxidant capacity (T-AOC) was observed in response to the 1.95 % Thr diet. Liver proapoptotic factors (bax and caspase9) were significantly elevated in fish fed the 1.39 % Thr diet. The nuclear factor κB (nf-κb) and the NF-κB-mediated inflammatory cytokines were significantly increased in fish fed the 1.39 % Thr diet. The mRNA levels of protein kinase R (PKR)-like endoplasmic reticulum kinase (perk), eukaryotic translation initiation factor 2 (eif2α), activating transcription factor 4 (atf4), and C/EBP-homologous protein (chop), which are factors in the PERK/ATF4/CHOP pathway, one of the unfolded protein response (UPR) signaling pathways, were significantly increased by the 1.39 % Thr diet. The present study showed that low dietary Thr levels caused oxidative damage, reduced immunity, might affected PERK-ATF4-CHOP signaling to induce apoptosis, and via NF-κB signaling to trigger an inflammatory response in largemouth bass. Based on the TP, ALB, CAT, and MDA levels, the dietary Thr requirements for juvenile largemouth bass were estimated to be 2.18 %, 2.29 %, 1.92 %, and 1.88 %. Furthermore, these findings provided a new perspective that low dietary amino acid levels may trigger apoptosis and inflammatory responses in fish through the UPR signaling pathway.

Metabolomic-derived endotypes of age-related macular degeneration (AMD): a step towards identification of disease subgroups.

Age-related macular degeneration (AMD) is a leading cause of blindness worldwide, with a complex pathophysiology and phenotypic diversity. Here, we apply Similarity Network Fusion (SNF) to cluster AMD patients into putative metabolomics-derived endotypes. Using a discovery cohort of 163 AMD patients from Boston, US, and a validation cohort of 214 patients from Coimbra, Portugal, we identified four distinct metabolomics-derived endotypes with varying retinal structural and functional characteristics, confirmed across both cohorts. Patients clustered into Endotype 1 exhibited a milder form of AMD and were characterized by low levels of amino acids in specific metabolic pathways. Meanwhile, patients clustered into both Endotype 3 and 4 were associated with more severe AMD and exhibited low levels of fatty acid metabolites and elevated levels of sphingomyelins and fatty acid metabolites, respectively. These preliminary findings indicate that metabolomics-derived endotyping may offer a refined strategy for categorizing AMD patients based on their specific pathophysiological underpinnings, rather than relying solely on traditional observational clinical indicators.

Soluble Organic Compounds and Cyanide in Apollo 17 Lunar Samples: Origins and Curation Effects

AbstractWe analyzed 12 Apollo 17 samples through the Apollo Next Generation Sample Analysis (ANGSA) program to determine the abundances of a variety of compound classes, including amino acids, aldehydes, ketones, amines, carboxylic acids, and cyanide‐releasing species. Analyzed samples included portions of double drive tube 73001/73002, the bottom half of which (73001) was hermetically sealed under lunar vacuum, as well as lunar regolith samples from three different illumination environments that had been curated frozen for ∼50 years. Consistent with previous results, we detected low levels of amino acids in the hot‐water extracts of most samples (0.55–12.03 nmol/g in unhydrolyzed samples; 0.53–72.38 nmol/g after acid hydrolysis of the extracts). We also detected one‐carbon and two‐carbon species of amines, aldehydes, and carboxylic acids (i.e., methylamine, ethylamine, formaldehyde, acetaldehyde, formic acid, acetic acid) not previously reported in lunar samples, as well as insoluble cyanide‐forming species. Although these compounds are potential precursor molecules for amino acids, no specific precursor relationships could be determined. Nylon contamination was the likely source of some amino acids. Abundances of some species (e.g., cyanide) decreased with increasing depth in the drive tube, suggesting that exogenous delivery and concentration mechanisms near the surface outweighed surface degradation processes. In addition, we observed the potential cold trapping of volatile amines in the persistently shadowed samples analyzed. Finally, we noted the effects of different curation conditions; hermetic sealing appeared to preserve higher amounts of volatile compounds, while frozen curation did not have a noticeable preservation effect on the organic volatiles analyzed here.

Local Sources of Protein in Low- and Middle-Income Countries: How to Improve the Protein Quality?

Protein inadequacy is a major contributor to nutritional deficiencies and adverse health outcomes of populations in low- and middle-income countries (LMICs). People in LMICs often consume a diet predominantly based on staple crops, such as cereals or starches, and derive most of their daily protein intakes from these sources. However, plant-based sources of protein often contain low levels of indispensable amino acids (IAAs). Inadequate intake of IAA in comparison with daily requirements is a limiting factor that results in protein deficiency, consequently in the long-term stunting and wasting. In addition, plant-based sources contain factors such as antinutrients that can diminish protein digestion and absorption. This review describes factors that affect protein quality, reviews dietary patterns of populations in LMICs and discusses traditional and novel small- and large-scale techniques that can improve the quality of plant protein sources for enhanced protein bioavailability and digestibility as an approach to tackle malnutrition in LMICs. The more accessible small-scale food-processing techniques that can be implemented at home in LMICs include soaking, cooking, and germination, whereas many large-scale techniques must be implemented on an industrial level such as autoclaving and extrusion. Limitations and considerations to implement those techniques locally in LMICs are discussed. For instance, at-home processing techniques can cause loss of nutrients and contamination, whereas limitations with larger scale techniques include high energy requirements, costs, and safety considerations. This review suggests that combining these small- and large-scale approaches could improve the quality of local sources of proteins, and thereby address adverse health outcomes, particularly in vulnerable population groups such as children, adolescents, elderly, and pregnant and lactating women.

Unveiling Disrupted Lipid Metabolism in Benign Prostate Hyperplasia, Prostate Cancer, and Metastatic Patients: Insights from a Colombian Nested Case-Control Study.

Prostate cancer is a significant global health concern, and its prevalence is increasing worldwide. Despite extensive research efforts, the complexity of the disease remains challenging with respect to fully understanding it. Metabolomics has emerged as a powerful approach to understanding prostate cancer by assessing comprehensive metabolite profiles in biological samples. In this study, metabolic profiles of patients with benign prostatic hyperplasia (BPH), prostate cancer (PCa), and metastatic prostate cancer (Met) were characterized using an untargeted approach that included metabolomics and lipidomics via liquid chromatography and gas chromatography coupled with high-resolution mass spectrometry. Comparative analysis among these groups revealed distinct metabolic profiles, primarily associated with lipid biosynthetic pathways, such as biosynthesis of unsaturated fatty acids, fatty acid degradation and elongation, and sphingolipid and linoleic acid metabolism. PCa patients showed lower levels of amino acids, glycerolipids, glycerophospholipids, sphingolipids, and carnitines compared to BPH patients. Compared to Met patients, PCa patients had reduced metabolites in the glycerolipid, glycerophospholipid, and sphingolipid groups, along with increased amino acids and carbohydrates. These altered metabolic profiles provide insights into the underlying pathways of prostate cancer's progression, potentially aiding the development of new diagnostic, and therapeutic strategies.

Influence of yeast growth conditions and proteolytic enzymes on the amino acid profiles of yeast hydrolysates: Implications for taste and nutrition

This study investigated the effects of aerobic and anaerobic growth and proteolytic enzymes on the amino acid content of yeast hydrolysates in relation to taste and nutrition. Saccharomyces cerevisiae ATCC5574 was grown under fed-batch aerobic or batch anaerobic conditions. Intracellular glutamic acid (Glu) concentrations were 18-fold higher in aerobic yeast.Hydrolysis with papain and alkaline protease released more amino acids (AA) than simple autolysis or hydrolysis with bromelain, most significantly when applied to aerobic yeast (∼2-fold increase). Autolysates and bromelain hydrolysates from aerobic yeast had low levels of bitter and essential AAs, with high levels of umami Glu. Papain and alkaline protease hydrolysates of aerobic yeast had high levels of umami, bitter and essential AAs. Autolysates/hydrolysates from anaerobic yeast had moderate, high, and low levels of bitter, essential and umami AAs.Selection of both yeast growth conditions and hydrolysis enzyme can manipulate the free AA profile and yield of hydrolysates.

Metabolites for monitoring symptoms and predicting remission in patients with depression who received electroconvulsive therapy: a pilot study

The lack of biomarkers to monitor and predict the efficacy of electroconvulsive therapy (ECT) has hindered its optimal use. To establish metabolomic markers for monitoring and predicting the treatment efficacy of ECT, we comprehensively evaluated metabolite levels in patients with major depressive disorder (MDD) by performing targeted and non-targeted metabolomic analyses using plasma samples before and after the first, third, and final ECT sessions, and 3–7 days after the final session. We compared the plasma metabolomes of age- and sex-matched healthy controls (HCs). Thirteen hospitalized patients with MDD and their corresponding HCs were included in this study. We observed that patients with MDD exhibited lower levels of amino acids, including gamma-aminobutyric acid (GABA), and metabolites involved in tryptophan metabolism and the kynurenine pathway, and higher levels of cortisol at baseline. Furthermore, we investigated the relationship between metabolite levels and depression severity across seven measurement timepoints along with one correlation analysis and found that amino acids, including GABA and tryptophan catabolites, were significantly correlated with the severity of depression. Despite the exploratory nature of this study due to the limited sample size necessitating further validation, our findings suggest that the blood metabolic profile has potential as a biomarker for ECT.

Variations in Total Protein and Amino Acids in the Sequenced Sorghum Mutant Library.

Sorghum (Sorghum bicolor) is the fifth most important cereal crop worldwide; however, its utilization in food products can be limited due to reduced nutritional quality related to amino acid composition and protein digestibility in cooked products. Low essential amino acid levels and digestibility are influenced by the composition of the sorghum seed storage proteins, kafirins. In this study, we report a core collection of 206 sorghum mutant lines with altered seed storage proteins. Wet lab chemistry analysis was conducted to evaluate the total protein content and 23 amino acids, including 19 protein-bound and 4 non-protein amino acids. We identified mutant lines with diverse compositions of essential and non-essential amino acids. The highest total protein content in these lines was almost double that of the wild-type (BTx623). The mutants identified in this study can be used as a genetic resource to improve the sorghum grain quality and determine the molecular mechanisms underlying the biosynthesis of storage protein and starch in sorghum seeds.

1-Methylcyclopropene delays degradation of peel greenness but induces internal physiological disorders in cold-stored fruit of interspecific pears

The green-skinned interspecific ‘Greensis’ pear cultivar is a promising pear cultivar due to its crispness, juiciness, and strong resistance to scab disease. Postharvest storability and metabolic activity can be maintained by treating cold-stored pears with 1 μL L−1 of 1-methylcyclopropene (1-MCP) for 16 hr at 22 °C. After 1-MCP treatment, the ‘Greensis’ pears were stored at 0.5 °C for up to 6 months, after which they were transferred at 22 °C for 7 days of shelf life. 1-MCP treatment had no visible effects on physiological responses, such as fruit weight loss, firmness, titratable acidity, and soluble solids content. However, metabolic profile analyses indicated that the 1-MCP-treated fruit had lower levels of amino acids and soluble carbohydrates (except for sucrose) than the control fruit. The incidence of cortex browning and cavities was much higher in 1-MCP-treated fruit than in untreated fruit. Similarly, the chroma values of the peel and cortex tissues were higher in 1-MCP-treated fruit than in untreated fruit. Moreover, the total chlorophyll and chlorophyll a contents were more preserved in 1-MCP-treated fruit than in untreated pears. Overall, 1-MCP treatment increased sucrose, chlorophyll a, and total chlorophyll contents, thus delaying chlorophyll degradation during storage. These results suggested that 1-MCP treatment delayed the skin color degradation of interspecific ‘Greensis’ pears during storage and prevented changes in the profiles of major metabolites. Nevertheless, the incidence and severity of cortex browning and cavities were highly increased in 1-MCP-treated fruit during storage.

Comparative assessment of metabolic, ionic and molecular responsiveness of four facultative halophytes to habitat salinization in the southwest of Jeddah Governorate, Saudi Arabia.

This study explores the influence of salinity on some physiological and biochemical pathways of four facultative halophytes (Abutilon pannosum, Indigofera oblongifolia, Senna italica, and Tetraena coccinea) along the southwest coast of Jeddah Governorate. Through a comparative analysis of these plants in both saline and non-saline environments, the study investigates chlorophyll levels, ion concentrations within the plants, the correlation with the SOS1 gene, and the impact of salinity on metabolic compounds. The overarching goal is to gain insights into the adaptive mechanisms of these specific plants to salt stress, providing valuable information for addressing global agricultural challenges associated with salinity. Throughout the study, metabolic, ionic, and molecular responses of these plants were scrutinized in both environments. The findings revealed elevated levels of Na+, K+, Ca2+, and Mg2+ in saline habitats, except for Na+ in I. oblongifolia. Despite increased concentrations of Chl b, variations were noted in Chl a and carotenoids in plants exposed to salt. Osmoregulatory patterns in A. pannosum and I. oblongifolia exhibited reversible changes, including heightened protein and proline levels in A. pannosum and decreased levels in I. oblongifolia, accompanied by alterations in amino acids and soluble carbohydrates. Senna italica displayed higher levels of osmolytes, excluding proline, compared to salinized environments, while T. coccinea exhibited lower levels of amino acids. The accumulation of Na+ emerged as the primary mechanism for ionic homeostasis in these plants, with non-significant decreases observed in K+, Mg2+, and Ca2+. Notably, an overexpression of the SOS1 gene (plasma membrane Na+/H+ antiporter) was observed as a response to maintaining ionic balance. Understanding these halophytes will be critical in addressing salinity challenges and enhancing crop tolerance to salinity.

A multi-omics based anti-inflammatory immune signature characterizes long COVID-19 syndrome

To investigate long COVID-19 syndrome (LCS) pathophysiology, we performed an exploratory study with blood plasma derived from three groups: 1) healthy vaccinated individuals without SARS-CoV-2 exposure; 2) asymptomatic recovered patients at least three months after SARS-CoV-2 infection and; 3) symptomatic patients at least 3months after SARS-CoV-2 infection with chronic fatigue syndrome or similar symptoms, here designated as patients with long COVID-19 syndrome (LCS). Multiplex cytokine profiling indicated slightly elevated pro-inflammatory cytokine levels in recovered individuals in contrast to patients with LCS. Plasma proteomics demonstrated low levels of acute phase proteins and macrophage-derived secreted proteins in LCS. High levels of anti-inflammatory oxylipins including omega-3 fatty acids in LCS were detected by eicosadomics, whereas targeted metabolic profiling indicated high levels of anti-inflammatory osmolytes taurine and hypaphorine, but low amino acid and triglyceride levels and deregulated acylcarnitines. A model considering alternatively polarized macrophages as a major contributor to these molecular alterations is presented.

Preference and performance of the green peach aphid, Myzus persicae on three Brassicaceae vegetable plants and its association with amino acids and glucosinolates.

The green peach aphid, Myzus persicae (Sulzer) is a generalist pest of various host plants, whose feeding preference and growth performance mainly depends on the quantity and quality of nutrients and defensive metabolites in host plants. Here, we studied the preference and performance of M. persicae on three major Brassicaceae vegetables in China and measured nutrient (amino acids) and defensive metabolites (glucosinolates) in these plants. We found that M. persicae preferred and performed better on Chinese cabbage than cabbage and radish, which may be due to the relatively higher concentration of amino acids and lower levels of indole glucosinolates in their leaves. The glucosinolates level in cabbage leaves was ten times higher than the other two plants, while the amino acid concentration in radish was only half of the cabbage or Chinese cabbage. The higher concentration of indole glucosinolates in cabbage and lower levels of amino acids in radish may account for the poorer preference and growth of M. persicae on these two plants. These results suggest that both amino acids and glucosinolates in plants may play important roles in the preference and performance of M. persicae, which provide new knowledge for the cultivation and breeding of Brassicaceae vegetables.

Pre-existing biochemical entities predict severity of cytokine release syndrome in B-cell lymphoma patients treated with CD19-directed CAR T-cells

Abstract Potentially severe immune-related adverse events (irAEs) develop in relapsed/refractory large B cell lymphoma (R/R LBCL) patients during treatment with anti-CD19 CAR T-cell therapy. Therefore, prediction and better management of toxicities is critically required to improve patient outcomes. Cytokine release syndrome (CRS), one of the most notable acute irAEs, is the result of an inflammatory response of activated CAR T-cells and myeloid cells. Interestingly, high levels of glucose and low levels of certain amino acids are associated with pro-inflammatory immune activation in infectious and autoimmune diseases, but such associations have not been investigated in CAR T-cell therapy. Therefore, we employed mass spectrometry-based untargeted and targeted metabolomics analysis to identify pre-treatment host biological metabolites that predict rapid onset and severe CRS in 41 R/R LBCL patients treated with Axicabtagene Ciloleucel or Tisagenlecleucel. Analysis of plasma metabolites revealed significant associations of CRS with several metabolic classes, including carbohydrates, lipid, amino acids, dipeptides and nucleotides. Interestingly, high levels of the carbohydrates, glucose, mannose and 1,5-anhydroglucitol were observed in patients who experienced more rapid onset of CRS. In contrast, patients with greater severity of CRS had lower plasma abundance of the amino acids, glycine, proline and glutamate. Our data suggest that specific pre-existing plasma metabolites have the potential to serve as predictive biomarkers of CRS in patients undergoing anti-CD19 CAR T-cell therapy, and may be used for risk stratification and clinical management of irAEs in advance of treatment.

A low amino acid environment promotes cell macropinocytosis through the YY1-FGD6 axis in Ras-mutant pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC), cancer with a high mortality rate and the highest rate of KRAS mutation, reportedly internalizes proteins via macropinocytosis to adapt to low amino acid levels in the tumor microenvironment. Here, we aimed to identify a key regulator of macropinocytosis for the survival of tumor cells in a low amino acid environment in PDAC. FYVE, RhoGEF, and PH domain-containing protein 6 (FGD6) were identified as key regulators of macropinocytosis. FGD6 promoted PDAC cell proliferation, macropinocytosis, and tumor growth both in vitro and in vivo. The macropinocytosis level was decreased with FGD6 knockdown in PDAC cell lines. Moreover, FGD6 promoted macropinocytosis by participating in the trans-Golgi network and enhancing the membrane localization of growth factor receptors, especially the TGF-beta receptor. TGF-beta enhanced macropinocytosis in PDAC cells. Additionally, YAP nuclear translocation induced by a low amino acid tumor environment initiated FGD6 expression by coactivation with YY1. Clinical data analysis based on TCGA and GEO datasets showed that FGD6 expression was upregulated in PDAC tissue, and high FGD6 expression was correlated with poor prognosis in patients with PDAC. In tumor tissue from KrasG12D/+/Trp53R172H/-/Pdx1-Cre (KPC) mice, FGD6 expression escalated during PDAC development. Our results uncover a previously unappreciated mechanism of macropinocytosis in PDAC. Strategies to target FGD6 and growth factors membrane localization might be developed for the treatment of PDAC.

Whole Genome Sequencing: Early Diagnostic Tool in Newborns with Refractory Seizures.

Whole Genome Sequencing: Early Diagnostic Tool in Newborns with Refractory Seizures.

Metabolomics in Placental Tissue from Women Living with HIV.

It is unknown whether antiretroviral (ARV) drugs in women living with HIV (WLHIV) are associated with mitochondrial toxicity and altered fat oxidation and branched-chain amino acid metabolism in the placenta and fetus. Immediately after delivery, we froze placental biopsies from 20 WLHIV and 20 matched uninfected women. We analyzed global biochemical profiles using high-performance liquid chromatography/tandem mass spectrometry and gas chromatography/mass spectrometry. We used t-tests, principle component analysis, hierarchical clustering, and random forest analysis (RFA) in our analysis. Twelve WLHIV were on protease inhibitors, six on non-nucleoside reverse inhibitors, and two on integrase strand inhibitors with optimized backbone. Mean birth weight of HIV-exposed neonates was significantly lower than unexposed neonates (3,075 g vs. 3,498 g, p = .01) at similar gestational age. RFA identified 30 of 702 analytes that differentiated the placental profiles of WLHIV from uninfected women with 72.5% predictive accuracy. Placental profiles of non-nucleoside reverse transcriptase inhibitor (NNRTI)-treated WLHIV exhibited lower levels of amino acids, including essential and branched-chain amino acids, and some medium-chain acylcarnitines. Placental metabolism may be altered in WLHIV, possibly associated with ARV exposure. The lower birth weight among neonates of WLHIV suggests the need for further studies considering potential deleterious effects of altered placenta metabolism on fetal growth and development.

The Impact of Photorespiratory Glycolate Oxidase Activity on Arabidopsis thaliana Leaf Soluble Amino Acid Pool Sizes during Acclimation to Low Atmospheric CO2 Concentrations.

Photorespiration is a metabolic process that removes toxic 2-phosphoglycolate produced by the oxygenase activity of ribulose-1,5-bisphosphate carboxylase/oxygenase. It is essential for plant growth under ambient air, and it can play an important role under stress conditions that reduce CO2 entry into the leaf thus enhancing photorespiration. The aim of the study was to determine the impact of photorespiration on Arabidopsis thaliana leaf amino acid metabolism under low atmospheric CO2 concentrations. To achieve this, wild-type plants and photorespiratory glycolate oxidase (gox) mutants were given either short-term (4 h) or long-term (1 to 8 d) low atmospheric CO2 concentration treatments and leaf amino acid levels were measured and analyzed. Low CO2 treatments rapidly decreased net CO2 assimilation rate and triggered a broad reconfiguration of soluble amino acids. The most significant changes involved photorespiratory Gly and Ser, aromatic and branched-chain amino acids as well as Ala, Asp, Asn, Arg, GABA and homoSer. While the Gly/Ser ratio increased in all Arabidopsis lines between air and low CO2 conditions, low CO2 conditions led to a higher increase in both Gly and Ser contents in gox1 and gox2.2 mutants when compared to wild-type and gox2.1 plants. Results are discussed with respect to potential limiting enzymatic steps with a special emphasis on photorespiratory aminotransferase activities and the complexity of photorespiration.

Metabolomics Reveals Differences in Aqueous Humor Composition in Patients With and Without Pseudoexfoliation Syndrome

Pseudoexfoliation syndrome (XFS) is stress- or inflammation-induced elastosis accompanied by excessive production of microfibrils and their deposition in the anterior segment of the eye. Approximately 60–70 million people are affected by XFS worldwide. It is a component of a systemic disorder, considered a major risk factor for accelerated cataract formation, cataract surgery complications and development of glaucoma, which untreated or inadequately treated may lead to blindness. Moreover, XFS has been associated with cardiovascular and cerebrovascular morbidity, dementia, sensorineural hearing loss and pelvic organ prolapse. The pathogenesis of XFS has not been fully elucidated yet. Aqueous humor (AH) is a transparent fluid filling the anterior and posterior chambers of the eye. Determination of AH metabolites that are characteristic for XFS may provide valuable information about the molecular background of this ocular disorder. The aim of this study was to compare the composition of AH in XFS and non-XFS patients undergoing cataract surgery. The AH samples from 34 patients (15 with XFS and 19 without) were analyzed using liquid chromatography coupled to a Quadrupole Time-of-Flight mass spectrometer (LC-QTOF-MS). The obtained metabolic fingerprints were analyzed using multivariate statistics. Eleven statistically significant metabolites were identified. Compared with the non-XFS group, the AH of patients with XFS contained significantly lower levels of amino acids and their derivatives, for example, arginine (−31%, VIP = 2.38) and homo-arginine (−19%, VIP = 1.38). Also, a decrease in the levels of two acylcarnitines, hydroxybutyrylcarnitine (−29%, VIP = 1.24) and decatrienoylcarnitine (−46%, VIP = 1.89), was observed. However, the level of indoleacetaldehyde in XFS patients was significantly higher (+96%, VIP = 2.64). Other significant metabolites were two well-recognized antioxidants, ascorbic acid (−33%, VIP = 2.11) and hydroxyanthranilic acid (−33%, VIP = 2.25), as well as S-adenosylmethionine, a compound with anti-inflammatory properties (−29%, VIP = 1.93). Metabolic pathway analysis demonstrated that the identified metabolites belonged to eight metabolic pathways, with cysteine and methionine metabolism as well as arginine and proline metabolism being the most frequently represented. XFS can be associated with enhanced oxidative stress and inflammation, as well as with the disturbances of cellular respiration and mitochondrial energy production. Implementation of non-targeted metabolomics provided a better insight into the still not fully understood pathogenesis of XFS.