Asparagine Levels Research Articles

Aldehyde-mediated inhibition of asparagine biosynthesis has implications for diabetes and alcoholism.

Patients with alcoholism and type 2 diabetes manifest altered metabolism, including elevated aldehyde levels and unusually low asparagine levels. We show that asparagine synthetase B (ASNS), the only human asparagine-forming enzyme, is inhibited by disease-relevant reactive aldehydes, including formaldehyde and acetaldehyde. Cellular studies show non-cytotoxic amounts of reactive aldehydes induce a decrease in asparagine levels. Biochemical analyses reveal inhibition results from reaction of the aldehydes with the catalytically important N-terminal cysteine of ASNS. The combined cellular and biochemical results suggest a possible mechanism underlying the low asparagine levels in alcoholism and diabetes. The results will stimulate research on the biological consequences of the reactions of aldehydes with nucleophilic residues.

O efeito da toxemia na gravidez nos níveis séricos de carnitina e aminoácidos em cabras

ABSTRACT The aim of this study was to determine changes in the carnitine and amino acid profile of goats with clinical pregnancy toxemia. The study included a total of 40 Aleppo breed goats, 20 with clinical pregnancy toxemia and 20 healthy goats. The goats with low glucose level and BHBA of 1.6-5mmol/L formed the group with clinical pregnancy toxemia, and goats with high glucose level and BHBA <0.8mmol/L formed the control group. Carnitine and amino acid profiles were determined in the groups. The results showed that the serum BHBA level was significantly higher, and the glucose level was lower in the clinical pregnancy toxemia group (p<0.001). From the serum amino acid profiles, the levels of Methyl Glutaryl, Valine, Methionine, Phenylalanine, Tyrosine, Asparagine, Arginine, Glutamic Acid, Alanine and Ornithine were determined to be statistically significantly low in the pregnancy toxemia group (p<0.01), and lysine was determined to be significantly high (p<0.01). It was determined that all serum carnitine profiles, except Decenoylcarnitine and Propionylcarnitine, were higher in goats with clinical pregnancy toxemia (p<0.05). In conclusion, it was concluded that these two profiles can be used as biomarkers in the diagnosis of pregnancy toxemia.

Untargeted UHPLC-MS deferential screening method and targeted UHPLC-MS/MS quantitative method for the determination of amino acids in mouse brain tissue

An untargeted UHPLC-MS method was developed for the preliminary screening of differential amino acids between two mouse brain tissue samples, and a targeted UHPLC-MS/MS method was developed for the quantitative determination of amino acids in mouse brain tissue samples. The sample processing procedure, chromatographic conditions and MS parameters were optimized. The mouse brain tissue samples were homogenized with HCl solution (0.01 mol/L), protein precipitated with methanol and centrifugation. For the untargeted method, one sample was derivatized with butanol (D0)-HCl and another sample was derivatized with deuterated butanol (D9)-HCl, then combined together and analyzed with UHPLC-MS. Chromatographic peak area ratio for each amino acid was then used for differential analyses between two samples. Results showed good accuracy with standard spiked samples (83.4–113.0 %). For the targeted quantitative method, the sample supernatants were dried and residues were derivatized with butanol (D0)-HCl. Internal standards of amino acids derivatized with deuterated butanol (D9)-HCl were added before analyzed with UHPLC-MS/MS. The intra- and inter-day precisions were in the intervals of 0.5–8.1 % and 0.3–9.1 %. The LOD and the LOQ were ranged from 0.03 × 10−3 to 0.92 × 10−3 µg mL−1 and 0.08 × 10−3 to 3.08 × 10−3 µg mL−1, respectively. Spiked recoveries were from 96.8 % to 111.7 %. Both methods were applied to analyze amino acids in brain tissue of C57BL/6 mice treated and untreated with piperine and the result showed that the levels of glutamic acid, aspartic acid, histidine, asparagine, γ-aminobutyric acid, alanine, leucine and isoleucine significantly decreased in the piperine treated group comparing with the untreated group.

Association Between Human Metabolomics and Rheumatoid Arthritis: A Systematic Review and Meta-analysis

The underdiagnosis and inadequate treatment of rheumatoid arthritis (RA) can be attributed to the various clinical manifestations presented by patients. To address this concern, we conducted an extensive review and meta-analysis, focusing on RA-related metabolites. A comprehensive literature search was conducted in PubMed, the Cochrane Library, Web of Science, and Embase to identify relevant studies published up to October 5, 2022. The quality of the included articles was evaluated and, subsequently, a meta-analysis was conducted using Review Manager software to analyze the association between metabolites and RA. Forty nine studies met the inclusion criteria for the systematic review, and six of these studies were meta-analyzed to evaluate the association between 28 reproducible metabolites and RA. The results indicated that, compared to controls, the levels of histidine (RoM=0.83, 95% CI=0.79-0.88, I2=0%), asparagine (RoM=0.83, 95% CI=0.75-0.91, I2=0%), methionine (RoM=0.82, 95% CI=0.69-0.98, I2=85%), and glycine (RoM=0.81, 95% CI=0.67-0.97, I2=68%) were significantly lower in RA patients, while hypoxanthine levels (RoM=1.14, 95% CI=1.09-1.19, I2=0%) were significantly higher. This study identified histidine, methionine, asparagine, hypoxanthine, and glycine as significantly correlated with RA, thus offering the potential for the advancement of biomarker discovery and the elucidation of disease mechanisms in RA.

Assimilation of Nitrate into Asparagine for Transport in Soybeans

In this study, the systematic analysis of nitrate assimilation and transport in soybean roots was further improved by analyzing the concentrations of nitrate assimilates, asparagine and glutamine, in soybean roots and the related enzyme activities. This provided a theoretical basis for the efficient utilization of nitrogen fertilizer in soybean farming. A dual-root soybean system with both sides being nodulated was used to provide nitrate withdrawal and resupply in three phases on one side, while the other side received nutrients without nitrogen under sand culture conditions. Measurements were taken of the root’s nitrate reductase and glutamine synthetase activities, as well as the concentrations of asparagine. Measurements were also taken of the asparagine concentration in the basal root peeled skin (where the primary transport pathway is the phloem) and the woody parts (where the primary transport pathway is the xylem). Furthermore, the concentration of glutamine in the roots was also assessed. The findings indicated a positive correlation between the nitrate concentration in the roots and the activity of glutamine synthetase in the roots on both sides. The levels of asparagine in the roots, specifically those in the basal root peeled skin and woody part on both sides, rose when nitrate was added and declined when nitrate was removed. There was no significant change in glutamine concentration within the roots of soybeans. Hence, we deduced that the local provision of nitrate to the dual-root soybeans might enhance the absorption of nitrate into the roots on both sides. Additionally, the assimilated substances were predominantly carried as asparagine through the peeled skin and woody sections of the basal root, facilitating transportation in both directions within the soybean plants (from shoot to root and from root to shoot).

Overcoming Venetoclax (Ven) Resistance through Glutamine (Gln) Depletion: Final Analysis of the Phase 1 Trial of Ven and Pegcrisantaspase (PegC) Combination in Relapsed and Refractory (R/R) Acute Myeloid Leukemia (AML)

Background: Ven-based therapies have been an advancement in the front-line treatment of AML; however development of resistance is common and the efficacy of Ven-containing regimens in R/R AML has been far less promising. We reported in vitro and in vivo depletion of Gln induced by long-acting Erwinia asparaginase, PegC, inhibited proliferation of complex karyotype (CK) AML and synergistically enhanced the antiapoptotic activity of Ven-mediated antagonism of Bcl-2 by decreasing the expression of proteins such as Mcl-1, whose translation is cap-dependent. Here we report the final results of our phase 1 clinical trial (NCT04666649) of VenPegC for treatment of R/R AML including patients (pts) previously treated with Ven. Methods: Pts received Ven 400 mg on D1-28 and PegC (500, 750, 1000 IU/m 2 in cohorts 1-3, IV) D1 & D15 every 4 weeks (wk). Primary study endpoints were incidences of regimen limiting toxicities (RLT) and RP2D. Secondary endpoints included rate of composite complete remissions. Correlative studies included monitoring plasma amino acid levels and asparaginase activity, and expression of proteins regulating cap-dependent mRNA translation in bone marrow (BM) mononuclear cells pre- and post-VenPegC treatment. Results: As of July 31, 2023, 26 pts were enrolled of whom 24 received at least 1 dose of PegC and 17 completed at least 1 cycle of PegC (2 doses). The median age was 61 (range 24-79), 16 (62%) were female; 9 (35%) had CK-AML and 5 (19%) had TP53 mutations; 18 (69%) were previously treated with Ven. Median prior lines of treatment were 2 (range 1-5). No RLT was observed in cohort 1 (n=3). Cohort 2 was expanded to 6 pts due to transient asymptomatic hypertriglyceridemia. Two pts in cohort 3 (n=3) developed grade 3 hyperbilirubinemia determined as RLT. Expansion cohort included 11 pts who were treated at MTD/RP2D (PegC 750 IU/m 2 plus Ven 400 mg) given as out-pt without a need for hospitalization. No pancreatitis, thrombohemorrhagic events, or silent inactivation were observed in the entire study population. Overall response rate (ORR) for pts who received ≥1 cycle of PegC was 47% (8/17), Fig 1. CR+ CRh+CRi rate was 29% (5/17); 1 pt had a partial remission with full count recovery and is currently receiving cycle 6; 2 pts had stable disease. Four pts showed hematologic improvement. Twelve (71%) pts who received ≥1 cycle of PegC were previously treated with Ven and were Ven-refractory. ORR in Ven-refractory pts was 42% (5/12); of the 5 pts who harbored a RAS or PTPN11 mutation and previously treated with Ven, 60% responded. Remarkably, all pts (3/3) who had a RUNX1 mutation achieved a CR; Pt 6 (relapsed AML with NRAS/cKIT/RUNX1 mutations) achieved an MRD-negative CRh and underwent an allogeneic stem cell transplant (alloHSCT); Pt 12 (primary refractory AML with TP53/NRAS/RUNX1 mutations) achieved a clearance of BM blasts after 1 dose of PegC and achieved MRD-negative CR at the end of cycle 1, and Pt 15 (post-alloHSCT relapsed AML with NRAS/PTPN11/CSF3R/RUNX1 mutations) achieved a CRi and received 10 cycles of VenPegC. Pt 31 (post-alloHSCT relapsed AML with FLT3-ITD) who had persistent disease after gilteritinib achieved an MRD-negative CRi, pending 2 nd alloHSCT. Nadir plasma asparaginase activity of >0.1 IU/mL was achieved in cycle 1 for all pts who received ≥1 cycle of VenPegC. All pts with available results achieved undetectable asparagine (Asn) level by day 7 and Asn remained undetectable throughout the study. Plasma Gln reduction from baseline occurred in all pts (except 1) within cycle 1. Pts in cohort 1 reached nadir Gln after initial PegC exposure in an average of 22 days compared to 16 days in cohort 2. In general, pts who achieved CR had the lowest absolute plasma Gln levels. Available Glu and Gln are shown in Fig.1 for pts who achieved a CR/PR. Out of the 8 assessed pts (4 responders & 4 non-responders), VenPegC inhibited cap-dependent translation after 1 cycle in responders, as evidenced by decreased phosphorylated eIF4E (active, pheIF4E) in 4/4 pts and decreased phosphorylated 4EBP1 (inactive, ph4EBP1), a negative regulator of eIF4E, in 2/4 pts. Conversely, pheIF4E and ph4EBP1 expression was increased in 3/4 and 4/4 of the non-responders, respectively (Fig.2). Conclusion: VenPegC is a novel regimen that can induce complete remission in some heavily pretreated R/R AML patients, even with previous exposure to Ven. Pts with RUNX1 mutation, whose translation depends on pheIF4E, may benefit further from this regimen.

Radiocarbon Flux Measurements Reveal Mechanistic Insight into Heat-Stress Induction of Nicotine Biosynthesis in Nicotiana attenuata.

The effect of high-temperature (HT) stress on nicotine biosynthesis in Nicotiana attenuata was examined. Nicotine content was measured in mature leaves, young sink leaves, and in roots from well-watered plants grown at 25 °C as controls and from plants exposed to 38 °C and 43 °C temperatures applied for 24, 48, 72, and 96 h duration. At 38 °C, all leaf nicotine levels were significantly less than control plants for up to 72 h exposure but rose sharply thereafter to levels significantly greater than controls with 96 h exposure. In contrast, plants exposed to 43 °C never exhibited a reduction in leaf nicotine content and showed an increase in content with just 48 h exposure. Using radioactive 11CO2 and 13NO3-, we found that HT stress reduced both CO2 fixation and nitrate uptake. Furthermore, radiocarbon flux analysis revealed that 'new' carbon partitioning (as 11C) into the 11C-radiolabeled amino acid (AA) pool was significantly reduced with HT stress as were yields of [11C]-aspartic acid, an important AA in nicotine biosynthesis, and its beta-amido counterpart [11C]-asparagine. In contrast, [12C]-aspartic acid levels appeared unaffected at 38 °C but were elevated at 43 °C relative to controls. [12C]-Asparagine levels were noted to be elevated at both stress temperatures. Since HT reductions in carbon input and nitrogen uptake were noted to impede de novo AA biosynthesis, protein degradation at HT was examined as a source of AAs. Here, leaf total soluble protein (TSP) content was reduced 39% with long exposures to both stress temperatures. However, Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) which was 41% TSP appeared unaffected. Altogether, these results support the theory that plant proteins other than Rubisco degrade at elevated temperatures freeing up essential AAs in support of nicotine biosynthesis.

Sodium pyruvate improves the plasma amino acid profile in rats with L-arginine-induced acute pancreatitis.

Plasma amino acid levels are altered upon many pathological conditions including acute pancreatitis. It is unclear whether amino acids can be used as specific biomarker of acute pancreatitis severity or recovery. Development of acute pancreatitis is associated with mitochondrial dysfunction and decreased cytosolic ATP level. Sodium pyruvate is considered as a potential treatment of pancreatitis due to its ability to sustain mitochondrial oxidative and ATP-productive capacity in vitro. This study investigated the effect of sodium pyruvate on pancreatic morphology and plasma amino acid levels in rats with acute pancreatitis. Acute pancreatitis in rats was induced by administration of L-arginine (5g/kg) Experimental treatment group received sodium pyruvate (1g/kg) for 4days. On day 8 of the experiment, animals were killed, blood was collected and plasma amino acid concentration was determined with high-performance liquid chromatography. Histological examination showed large areas of fibrosis in the pancreas of animals treated with L-arginine irrespectively of sodium pyruvate administration. Sodium pyruvate improved the plasma amino acid levels. Rats with acute pancreatitis had significantly lower levels of most essential and non-essential amino acids and increased glutamate and aspartate in plasma. Administration of sodium pyruvate completely or partially restored the levels of methionine, phenylalanine, tryptophan, leucine, isoleucine, aspartate, asparagine and ornithine levels, while increasing glutamine and serine to levels significantly higher than control. Plasma lysine, alanine, arginine and taurine remained unaffected in all experimental groups. Sodium pyruvate may be considered for use as a maintenance therapy in acute pancreatitis.

Correlation between microbial community succession and flavor substances during fermentation of Yongchuan Douchi

Yongchuan douchi (YCDC) is a traditional Chinese condiment famous for its distinct flavor and texture. This study utilized high-throughput sequencing to analyze the microbial community changes during YCDC fermentation and investigate the relationship between its physicochemical properties, flavor compounds, and microorganisms. The sequencing results revealed that Staphylococcus, Bacillus, Leuconostoc, Weissella, Streptophyta, Nocardiopsis, Aspergillus, Penicillium, Starmerella, and Zygosaccharomyces were the dominant bacteria during the YCDC fermentation process. The free amino acid (FAAs), glutamic acid (Glu), asparagine (Asp), and organic acid (citric and succinic acid) levels increased gradually throughout fermentation. Additionally, 29 key flavor compounds were identified, with lipids, alcohols, aldehydes, and acids significantly affecting the flavor profile. Network graph analysis demonstrated positive correlations between Staphylococcus, Escherichia_Shigella, Brevibacterium, and 1-octene-3-ol, ethyl isobutyrate, and benzaldehyde, respectively, contributing to light, fruity, and floral aromas. Moreover, bacterial genera exhibited stronger potential correlations with flavor compounds than fungal genera. These findings enhance the understanding of microbial community succession in YCDC and offer a theoretical basis for flavor strain selection.

Amino Acid Profile Identified In Plasma Has Predictive and Diagnostic Value in Heart Failure Patients

Background: Metabolic disruptions have effects on the pathogenesis and progression of heart failure (HF). Because of these disruptions are remarkable in HF patients, identity of new biomarkers is necessary to facilitate the determination of HF. In the present study we planned to evaluate the relationship between plasma amino acids and HF. Materials and methods: 40 patients with cardiomyopathy (CMP) and 40 healthy controls were included in this cross-sectional prospective study. A profile consisting of 26 amino acids was measured by liquid chromatography and mass spectrometry method (LC-MS) from blood samples taken from fasting plasma. Results: 5 amino acids (asparagine, beta-alanine, phenylalanine, tyrosine, and isoleucine) levels were higher in CMP patients. Glutamine, isoleucine, and glycine could effectively distinguish CMP and healthy controls. Isoleucine and beta-alanine may be potential biomarkers for CMP. Isoleucine/alloisoleucine, histidine/isoleucine, aspartic acid/isoleucine, and glycine/phenylalanine could predict CMP patients. Three differential pathways (phenylalanine metabolism, tyrosine metabolism, and phenylalanine, tyrosine, tryptophan biosynthesis) were found to be the underlying molecular metabolisms of HF. Conclusion: The present study demonstrating that fasting plasma amino acids were closely associated with HF. Monitoring these amino acids with LC-MS could help the diagnosis and predicting HF, and provide new diagnostic goals and curative practices.

Serum Amino Acid Profiling in Children with Autistic Spectrum Disorder: Insights from a Single-Center Study in Southern Romania.

The objective of this study was to analyze the serum amino acid profile in children diagnosed with autistic spectrum disorder (ASD) in southern Romania. The analysis aimed to provide insights into the underlying metabolic dysregulations associated with ASD. ASD is a neurodevelopmental disorder characterized by impaired social interaction, communication deficits, and restricted repetitive behaviors. Although the exact cause of ASD is largely unknown, recent evidence suggests that abnormalities in amino acid metabolism may contribute to its pathogenesis. Therefore, studying the amino acid profile in children with ASD could offer valuable information for understanding the metabolic disturbances associated with this complex disorder. This single-center study examined serum samples from children diagnosed with ASD, utilizing advanced analytical techniques to quantify the levels of different amino acids, amino acid derivatives, and amino acid-like substances. The results showed a lower level of taurine and a higher level of asparagine and leucine in the ASD group versus the control group. In the ASD group, we observed significant differences in tryptophan and alpha-aminobutyric acid levels based on age, with higher tryptophan levels in children older than 7 years when compared to children younger than 7 years; however, no significant correlations were found with the ASD group older than 7 years old. Additionally, younger children with ASD exhibited higher levels of alpha-aminobutyric acid than older children with ASD. The findings from this study contribute to the growing body of knowledge on the metabolic aspects of ASD, highlighting potential biomarkers and therapeutic targets for improving the management and treatment of ASD in children.

The Association between Macrosomia and Amino Acids' Levels in Maternal and Cord Sera: A Case-Control Study.

In the case-control study, 78 pairs of mothers and newborns were recruited from December 2016 to November 2019. Participants were divided into the macrosomia group (BW ≥ 4000 g, n = 39) and the control group (BW between 2500 g and 3999 g, n = 39) according to the birth weight (BW) of newborns. Maternal vein blood samples were collected before delivery and cord vein blood samples were collected after birth. The levels of amino acids in maternal and cord sera were measured by liquid chromatography and mass spectrometry (LC-MS/MS) in the year 2021. The difference in amino acid levels in maternal and cord sera between the two groups was compared, and the contribution of each amino acid to the difference between the two groups was analyzed. Unconditional logistic regression analysis was used to test the relationship between macrosomia and amino acids. In maternal serum during the antepartum, the levels of asparagine, glutamine, methionine, alanine, and threonine in the macrosomia group were higher but arginine was lower than that in the control group (p < 0.05). In cord serum, the levels of lysine, histidine, phenylalanine, arginine, tryptophan, valine, isoleucine, glutamate, tyrosine, and total essential amino acid (EAA) in the macrosomia group were lower while glutamine was higher than that in the control group (p < 0.05). The ratios of EAA, valine, threonine, methionine, tryptophan, and alanine in maternal serum to those in cord serum were higher, while the ratio of glutamine was lower in the macrosomia group (p < 0.05). Arginine and threonine in maternal serum and glutamate, glutamine, and histidine in cord serum were associated with macrosomia (p < 0.05). Most of the amino acid levels in the maternal sera of the macrosomia group are higher than those in the control group, while most of the amino acids' levels in the cord sera of the macrosomia group are lower than those in the control group. The ratios of some amino acids in maternal serum to those in cord serum were different between the two groups. Arginine and threonine in maternal serum and glutamate, glutamine, and histidine in cord serum are closely related to macrosomia.

Acrylamide formation in apple juice concentrates during storage

This study investigated the acrylamide formation in model systems, laboratory-scale produced apple juice concentrates obtained from different stages of process, and commercial apple juice concentrates at different Brix levels during the storage. Changes in the concentrations of reducing sugars, asparagine, 5-hydroxymethylfurfural, 3-deoxyglucosone, and acrylamide were monitored. In addition, Schiff bases of 3-deoxyglucosone and 5-hydroxymethylfurfural with asparagine were confirmed using high-resolution mass spectrometry in order to understand the contribution of intermediates to acrylamide formation. The results revealed that acrylamide formed in both acidic-aqueous model systems and laboratory-scale apple juice concentrates during production. The concentrations of 3-deoxyglucosone, 5-hydroxymethylfurfural, and acrylamide increased significantly (p<0.05) with the increase in Brix levels whereas asparagine levels decreased in commercial apple juice concentrates during storage. The concentration of acrylamide reached at the maximum level of 0.30 ± 0.02 µmol/kg dry matter in 70 °Bx of the concentrates. The findings showed that long-term storage of apple juice concentrates at high Brix levels contributes the acrylamide formation even in unsuitable systems such as acidic and high moisture medium. The confirmation of Schiff base of 3-deoxyglucosone and 5-hydroxymethylfurfural with asparagine demonstrated that the acrylamide formation was attributed to the accumulation of intermediates leading to the conversion of asparagine to acrylamide.

The Role of the Bile Microbiome in Common Bile Duct Stone Development.

Common bile duct (CBD) stones are a health concern for 10-20% of individuals with symptomatic gallstones, leading to health complications and placing a burden on healthcare systems. This study was initiated to investigate the changes in microbiome compositions and the metabolic signature associated with CBD stones. The research approach integrated taxonomic and functional data with metabolomics data, complemented by in vivo experiments. In a single tertiary healthcare institution, a total of 25 patients were enrolled who had undergone endoscopic retrograde cholangiopancreatography (ERCP) between February 2019 and January 2021. We harvested DNA from bile samples acquired from these individuals. The amplification of the bacterial 16S rRNA gene V3-V4 region was conducted through polymerase chain reaction (PCR), followed by sequencing. We utilized QIIME2 for a comprehensive data analysis. Furthermore, we performed a metabolomic analysis of the bile samples using nuclear magnetic resonance (NMR) spectroscopy. For the assessment of functional gene enrichment, we employed MetaboAnalyst 5.0. Lastly, we executed in vivo experiments on C57BL/6 mice and undertook histological examinations of tissue samples. Out of the 25 study subjects, 17 underwent ERCP due to CBD stones (the CBD stone group), while the remaining 8 had the procedure for different reasons (the non-CBD stone group). An alpha diversity analysis showed a significantly greater microbial diversity in the bile samples of the non-CBD stone group (p < 0.01), and a beta diversity analysis confirmed the greater microbial compositional abundance in the gut microbiomes in this group (p = 0.01). A taxonomic analysis revealed that the abundances of Enterococcaceae and Enterococcus were higher in the bile microbiomes of the CBD stone group. A metabolic profile analysis showed that the acetate, formate, and asparagine levels were higher in the CBD stone group. A pathway enrichment analysis showed the metabolic pathways (Arginine and Proline Metabolism, Aspartate Metabolism, Glycine, and Serine Metabolism, and Ammonia Recycling pathways) that were associated with these differences. Preclinical experiments demonstrated systemic inflammation and extracellular trap formation in the CBD stone group. Our study highlights the importance of biliary dysbiosis and bile metabolites, specifically acetate and formate, in CBD stone development and progression. These findings have implications for the development of diagnostic and therapeutic strategies using microbiomes for CBD stones.

Comparison of Amino Acid Profile, ACE Inhibitory Activity, and Organic Acid Profile of Cow and Goat Yogurts Produced with Lactobacillus acidophilus LA-5, Bifidobacterium animalis subsp. lactis BB-12, and Classical Yogurt Culture.

In this study, we aimed to produce a standard, more functional, and nutritious yogurt by using 5 different combinations of cow milk and goat milk and 2 types of starter cultures (classical yogurt culture and commercial probiotic culture). It was determined that the use of different milk types and different starter cultures in yogurt production had a statistically very significant effect (P < 0.01) on all physicochemical, microbiological, and biochemical properties. In addition, the storage period was effective on all parameters examined at varying rates. In the context, the use of goat milk in the experimental yogurt samples caused an increase in the ACE inhibitory activity values and the count of S. thermophilus, while the use of cow milk caused an increase in serum separation and pH values. On the other hand, serum separation, pH values, and ACE inhibitory activity and phenylalanine and leucine levels were found to be higher in the yogurts produced by using ABT-2 probiotic culture. It was observed that an increase in the levels of asparagine, aspartic acid, proline, and serine, as well as lactic acid, orotic acid, and citric acid, is higher in the yogurts produced by using classical yogurt culture. It has been concluded that the combination of goat milk and cow milk at different proportions and the use of probiotic culture together in yogurt production can produce yogurt that is more functional and richer in terms of organic compounds and essential amino acids.

Distinct evolution of type I glutamine synthetase in Plasmodium and its species-specific requirement

Malaria parasite lacks canonical pathways for amino acid biosynthesis and depends primarily on hemoglobin degradation and extracellular resources for amino acids. Interestingly, a putative gene for glutamine synthetase (GS) is retained despite glutamine being an abundant amino acid in human and mosquito hosts. Here we show Plasmodium GS has evolved as a unique type I enzyme with distinct structural and regulatory properties to adapt to the asexual niche. Methionine sulfoximine (MSO) and phosphinothricin (PPT) inhibit parasite GS activity. GS is localized to the parasite cytosol and abundantly expressed in all the life cycle stages. Parasite GS displays species-specific requirement in Plasmodium falciparum (Pf) having asparagine-rich proteome. Targeting PfGS affects asparagine levels and inhibits protein synthesis through eIF2α phosphorylation leading to parasite death. Exposure of artemisinin-resistant Pf parasites to MSO and PPT inhibits the emergence of viable parasites upon artemisinin treatment.

1601-P: Childhood Plasma Amino Acid Profiles Predict the 10-Year Risk of Nonalcoholic Fatty Liver Disease—The BCAMS Study

Background and Aims: Amino acid profiles have been proved a promising tool to identify novel biomarkers and insight of the mechanisms of metabolic diseases; however, date on youth was still limited, particularly nonalcoholic fatty liver disease (NAFLD). We aimed to prospectively investigate if there exist any association between childhood plasma amino acid profiles and adulthood NAFLD, and explore the early predictor and the underlying mechanism for the development of hepatic steatosis. Methods: A total of 307 subjects aged 12.1±3.0 years at baseline were enrolled in this study at 10-year follow-up, recruited from the Beijing Child and Adolescent Metabolic Syndrome (BCAMS) study cohort, including 99 NAFLD patients and 208 normal controls. Thirty-two amino acids (AAs) in fasting plasma at baseline were evaluated by liquid chromatograph-mass spectrometry. NAFLD was diagnosed by abdominal ultrasonography at follow-up. Results: Among the 32 qualified AAs, 14 AAs showed significant differences (p&amp;lt;0.05) between NAFLD and normal controls, including four AAs survived Bonferroni correction (p&amp;lt;0.0016). Adjustment for age, gender and lifestyle factors, there were 10 AAs remain significant, including alanine, asparagine, glutamine, glycine, isoleucine, tyrosine, serine and threonine etc. Further adjustment for BMI, lower levels of asparagine (OR: 0.62, 95% CI: 0.44-0.87), glutamine (OR: 0.73, 95% CI: 0.53-0.99) and serine (OR: 0.71, 95% CI: 0.51-0.98) remain significant predictors of NAFLD at the 10-year follow-up. Conclusion: Plasma amino acid profiles in childhood can predict risk of NAFLD in young adults. Plasma asparagine, glutamine and serine are likely to be early predictors of NAFLD independently of adiposity. Disclosure M.Li: None. S.Gao: None. Bcams group: n/a. Funding National High Level Hospital Clinical Research Funding (2022-PUMCH-C-014); National Natural Science Foundation of China (81970732, 82270924); Capital's Funds for Health Improvement and Research (2020-2Z-40117); CAMS Innovation Fund for Medical Sciences (C

A phase I dose escalation study of eryaspase in combination with modified FOLFIRINOX in locally advanced and metastatic pancreatic ductal adenocarcinoma.

e16288 Background: The prognosis for advanced metastatic pancreatic adenocarcinoma remains dismal, highlighting the need for novel therapeutic agents. FOLFIRINOX (5-Fluorouracil + Oxaliplatin + Irinotecan) remains the standard of care frontline therapy for advanced disease, with historical objective response rate (ORR) of ~31%. Eryaspase is an investigational red blood cell product encapsulating L-asparaginase, which hydrolyzes and reduces asparagine levels in plasma, leading to cancer cell death. A phase III trial (Trybeca-1), which compared chemotherapy with or without eryaspase in the second line setting, did not reach its primary efficacy endpoint. However, there was a trend towards improved survival in the group receiving 5-fluorouracil and irinotecan plus eryaspase. We herein report the final results from our phase I front-line study of mFOLFIRINOX plus eryaspase. Methods: Patients with locally advanced or metastatic biopsy-proven pancreatic adenocarcinoma were treated with mFOLFIRINOX plus eryaspase in a standard 3 +3 dose escalation trial design. mFOLFIRINOX was given as 5-Fluorouracil 2400 mg/m2 over 46 hours, Oxaliplatin 85 mg/m2, Irinotecan 150 mg/m2 plus eryaspase 75 units/kg at dose level 0 or 100 units/kg at dose level 1. Key eligibility criteria included performance status of 0 or 1, locally advanced or metastatic disease, and adequate organ function. The primary objectives were to determine the maximum tolerated dose (MTD) and to determine the safety of this combination. Results: At completion of trial, eighteen patients were enrolled. Three patients were enrolled at dose level 0 and fifteen at dose level 1, with no dose limiting toxicities. Seventeen patients had imaging to evaluate best response: 24% (N=4) had partial response (PR), 65% (N=11) had stable disease (SD), and 11% (N=2) had progressive disease. The disease control rate (PR +SD) was 89%. Progression free survival (PFS) was 6.4 months (95% CI 3.21 – 16.79) and overall survival (OS) was 10.1 months (95% CI 7.18 – NR). There were select patients with very durable response (PFS&gt;12 months and OS &gt;18 months). There was one grade 5 (G5) sepsis adverse event (AE) and one G4 thromboembolic AE. G3 AEs were hypokalemia (22%), fatigue (11%), anemia (6%), hypotension (6%), diarrhea (6%), syncope (6%), and atrial fibrillation (6%). Pharmacokinetic and pharmacodynamic data are in process. Conclusions: The novel combination of mFOLFIRINOX plus eryaspase was well tolerated with no DLT and demonstrated signals of clinical activity in select patients. The MTD was declared with 5-FU 2400 mg/2, Oxaliplatin 85 mg/2, Irinotecan 150 mg/m2, and eryaspase 100 units/kg. Although the phase III Trybeca-1 trial did not meet the primary OS endpoint, both trials may suggest enhanced activity of eryaspase with regimens containing 5-Fluorouracil and Irinotecan. Clinical trial information: NCT04292743 .

Quantification of Serum Metabolites in Early Colorectal Adenomas Using Isobaric Labeling Mass Spectrometry.

A major challenge in reducing the death rate of colorectal cancer is to screen patients using low-invasive testing. A blood test shows a high compliance rate with reduced invasiveness. In this work, a multiplex isobaric tag labeling strategy coupled with mass spectrometry is adopted to relatively quantify primary and secondary amine-containing metabolites in serum for the discovery of metabolite level changes of colorectal cancer. Serum samples from patients at different risk statuses and colorectal cancer growth statuses are studied. Metabolite identification is based on accurate mass matching and/or retention time of labeled metabolite standards. We quantify 40 metabolites across all the serum samples, including 18 metabolites validated with standards. We find significantly decreased levels of threonine and asparagine in the patients with growing adenomas or high-risk adenomas (p < 0.05). Glutamine levels decrease in patients with adenomas of unknown growth status or high-risk adenomas. In contrast, arginine levels are elevated in patients with low-risk adenoma. Receiver operating characteristic analysis shows high sensitivity and specificity of these metabolites for detecting growing adenomas. Based on these results, we conclude that a few metabolites identified here might contribute to distinguishing colorectal patients with growing adenomas from normal individuals and patients with unknown growth status of adenomas.

Impact of Some Enzymatic Treatments on Acrylamide Content in Biscuits

Since its discovery in many heat-treatment foods in 2002, many efforts have been made to reduce acrylamide levels in foods. Methods to reduce acrylamide levels by reducing Maillard reaction products have been considered. However, baking cookies produces acrylamide, a carcinogenic compound. This study aimed to use a new quantitative index and formula for L-asparaginase, glucose oxidase, their 1:1 blending enzymes, baker’s yeast, and green tea powder (0.5 g/kg wheat flour) at a new proposed temperature of 37 °C for 30 min to reduce acrylamide production in biscuits and bakery products using new indicators such as asparagine reduction (%), the asparagine/acrylamide ratio, acrylamide reduction (%), and the asparagine/reducing sugar ratio. The highest acrylamide concentrations were reduced from 865 mg/kg in the blank sample (BT0) to 260 and 215 mg/kg in the mixed enzyme powder (1:1) (BT3)- and BT4-treated samples, respectively. The biscuit samples treated with 0.5 g/kg L-asparaginase reduced the acrylamide levels by approximately 67.63%, while the BT3 samples showed acrylamide levels of 69.94% and asparagine levels of 68.75% and 47%, respectively, compared with percentage in the untreated sample (blank), 95%. This percentage was 54.16% for the BT4 samples. The results showed that acrylamide was formed during baking, and all treatment samples inhibited its formation, making it possible to produce foods with low levels of acrylamide in starchy foods in the food industry at 37 °C for 30 min and preserving the quality and nutritional value of the final product. It can be used as a specialty food or functional food and protects school-agechildren, as well as youth on campus, from approximately 70–80% of their daily intake of acrylamide.