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Background: Intrauterine growth restriction (IUGR) affects up to 30% of piglets in a litter. Piglets exposed to IUGR prioritize brain development during gestation, resulting in a higher brain-to-liver weight ratio (BrW/LW) at birth. IUGR is associated with increased mortality, compromised metabolism, and gut health. However, the dynamic metabolic and microbial shifts in IUGR-affected pigs remain poorly understood. This study aimed to investigate the longitudinal effects of IUGR, defined by a high BrW/LW, on the composition of faecal microbiota and plasma metabolome in pigs from birth to slaughter. One day (± 1) after birth, computed tomography was performed on each piglet to assess their brain and liver weights. The pigs with the highest (IUGR = 12) and the lowest (NORM = 12) BrW/LW were selected to collect faeces and blood during lactation (day 16 ± 0.6, T1) and at the end of the starter period (day 63 ± 8.6, T2) and faeces at the beginning (day 119 ± 11.4, T3) and end of the finisher period (day 162 ± 14.3, T4). Results: Faecal microbial Alpha diversity remained unaffected by IUGR across all time points. However, the Beta diversity was influenced by IUGR at T1 (P = 0.002), T2 (P = 0.08), and T3 (P = 0.03). Specifically, IUGR pigs displayed higher abundances of Clostridium sensu stricto 1 (Padj = 0.03) and Romboutsia (Padj = 0.05) at T1, Prevotellaceae NK3B31 group (Padj = 0.02), Rikenellaceae RC9 gut group (Padj = 0.03), and Alloprevotella (Padj = 0.03) at T2, and p-2534-18B5 gut group (Padj = 0.03) at T3. Conversely, the NORM group exhibited higher abundances of Ruminococcus (Padj = 0.01) at T1, HT002 (Padj = 0.05) at T2, and Prevotella_9 (Padj < 0.001) at T3. None of the plasma metabolites showed significant differences at T1 between the IUGR and NORM pigs. However, at T2, asparagine was lower in the IUGR compared to the NORM group (P < 0.05). Conclusions: These findings show that growth restriction in the uterus has a significant impact on the faecal microbiota composition in pigs, from birth to the beginning of the finisher period, but minimally affects the plasma metabolome profile. [ABSTRACT FROM AUTHOR]

We report on the dynamic interactions between β-cyclodextrin (β-CD) and each one of the two enantiomers of asparagine (d-Asp, l-Asp). Molecular docking methodologies were applied to elucidate the formation of the β-CD—d-Asp and β-CD—l-Asp inclusion complexes. Ultrasonic relaxation spectra revealed a single relaxation process in the frequency range studied that is attributed to the complexation between β-CD and asparagine enantiomers. Kinetic parameters and thermodynamic properties for each system were determined directly from the concentration- and temperature-dependent acoustic measurements, respectively. Both β-CD—d-Asp and β-CD—l-Asp systems revealed subtle differences in their thermodynamic and kinetic properties. The infrared absorption spectra of the host molecule, the guest enantiomers, and both inclusion complexes were recorded to verify and further elucidate the complexation mechanism. DFT methodologies were performed to calculate the theoretical IR spectra of the inclusion complexes and compared with the corresponding experimental spectra. The close resemblance between the experimental and theoretically predicted IR spectra is supportive of the formation of inclusion complexes. The encapsulation of asparagine enantiomers in β-cyclodextrin enables not only applications in drug delivery but also the detection and separation of chimeric molecules. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Several members of the NIMA-related kinase (NEK) family have been implicated in tumor progression; however, the role and underlying mechanisms of NEK8 in gastric cancer (GC) remain unclear. This study revealed a significant upregulation of NEK8 in GC, identifying it as an independent prognostic marker in patients with GC. Consistent with these findings, NEK8 silencing substantially impeded GC aggressiveness both in vitro and in vivo, while its overexpression produced the opposite effect. Gene Ontology enrichment analysis and metabolic profiling indicated that the impact of NEK8 on GC is primarily associated with reprogramming asparagine metabolism and modulating the mTORC1 pathway. Specifically, NEK8 knockdown suppressed asparagine synthesis by downregulating asparagine synthetase (ASNS) expression in GC cells. A strong correlation was observed between NEK8 levels and ASNS expression in human GC cells and tissue samples. Mechanistically, NEK8 directly interacts with ASNS, phosphorylating it at the S349 site, which inhibits its ubiquitination and subsequent degradation. Moreover, substituting the ASNS-S349 site with alanine abrogated the pro-tumorigenic effects of ASNS-WT overexpression. Additionally, asparagine was identified as an activator of the mTORC1 pathway, with reintroducing asparagine after NEK8 silencing restoring mTORC1 activity. Collectively, these findings demonstrate that NEK8-mediated asparagine synthesis and activation of the mTORC1 pathway play a critical role in promoting GC progression. [ABSTRACT FROM AUTHOR]

The aim of this study was to analyze the association between circulating amino acids and central and peripheral body temperature in subjects with and without fibromyalgia syndrome (FMS). A total of 47 patients with FMS and 59 healthy subjects were included in the study. The concentration of amino acids was determined in serum samples using a fluorimeter coupled with a high-performance liquid chromatography system. An infrared thermography camera was used to estimate peripheral hand temperatures. The core temperature of the body was estimated using an infrared thermometer, which was applied to the axillary and tympanic areas. Correlations between several thermographic variables of the hands and tryptophan, methionine, 3-methylhistidine, histidine, glutamic acid, and tyrosine were identified exclusively within the FMS group. In contrast, correlations between aminoadipic acid and serine and thermographic variables were observed only in the healthy control group. The concentrations of asparagine and lysine correlated with thermographic variables in both groups. The essential amino acid leucine was found to correlate with axillary temperature in FMS patients. However, it should be noted that the observed associations between aminoadipic acid and tryptophan blood concentrations and axillary temperature were limited to the control group. Several correlations were identified between circulating amino acids and different body temperatures in both healthy controls and patients with FMS. However, the correlation pattern differs significantly between FMS patients and healthy controls. These findings suggest the possibility of a change in the function of several amino acids in the thermoregulatory process in patients with FMS. [ABSTRACT FROM AUTHOR]

Many voltage-gated potassium (Kv) channels display a time-dependent phenomenon called C-type inactivation, whereby prolonged activation by voltage leads to the inhibition of ionic conduction, a process that involves a conformational change at the selectivity filter toward a non-conductive state. Recently, a high-resolution structure of a strongly inactivated triple-mutant channel kv1.2-kv2.1-3m revealed a novel conformation of the selectivity filter that is dilated at its outer end, distinct from the well-characterized conductive state. While the experimental structure was interpreted as the elusive non-conductive state, our molecular dynamics simulations and electrophysiological measurements show that the dilated filter of kv1.2-kv2.1-3m is conductive and, as such, cannot completely account for the inactivation of the channel observed in the structural experiments. The simulation shows that an additional conformational change, implicating isoleucine residues at position 398 along the pore lining segment S6, is required to effectively block ion conduction. The I398 residues from the four subunits act as a state-dependent hydrophobic gate located immediately beneath the selectivity filter. These observations are corroborated by electrophysiological experiments showing that ion permeation can be resumed in the kv1.2-kv2.1-3m channel when I398 is mutated to an asparagine--a mutation that does not abolish C-type inactivation since digitoxin (AgTxII) fails to block the ionic permeation of kv1.2-kv2.1-3m_ I398N. As a critical piece of the C-type inactivation machinery, this structural feature is the potential target of a broad class of quaternary ammonium (QA) blockers and negatively charged activators thus opening new research directions toward the development of drugs that specifically modulate gating states of Kv channels. [ABSTRACT FROM AUTHOR]

ABSTRACT Acrylamide (AA) and 5‐hydroxymethylfurfural (HMF) are the byproducts of the Maillard reaction that pose health risks due to their high exposure to the Western diet. Current study examines the impact of yeast fermentation on AA, HMF formation, and chemical composition of French fries. Fermentation, known for enriching food with beneficial probiotics, vitamins, and minerals, is coupled with blanching, a technique used to inactivate enzymes and reduce microbial load, thus preserving color and extending shelf life. Fermentation reduced HMF and AA levels in French fries by approximately 86.5% and 54.78%, respectively. Furthermore, combining fermentation with blanching treatment further decreased HMF and AA levels by 88.62% and 59.81%, respectively. This research suggests that yeast fermentation, particularly when combined with blanching treatment, can effectively mitigate the formation of AA and HMF in French fries, thereby potentially reducing their health risks. This pioneering research opens avenues for additional inquiries, laying the groundwork for future advancements in enhancing food safety. [ABSTRACT FROM AUTHOR]

Bacterial cancer therapy (BCT) is a promising therapeutic for solid tumours. Salmonella enterica Typhimurium (STm) is well-studied amongst bacterial vectors due to advantages in genetic modification and metabolic adaptation. A longstanding paradox is the redundancy of T cells for treatment efficacy; instead, STm BCT depends on innate phagocytes for tumour control. Here, we used distal T cell receptor (TCR) and IFNγ reporter mice (Nr4a3-Tocky-Ifnγ-YFP) and a colorectal cancer (CRC) model to interrogate T cell activity during BCT with attenuated STm. We found that colonic tumour infiltrating lymphocytes (TILs) exhibited a variety of activation defects, including IFN-γ production decoupled from TCR signalling, decreased polyfunctionality and reduced central memory (TCM) formation. Modelling of T-cell–tumour interactions with a tumour organoid platform revealed an intact TCR signalosome, but paralysed metabolic reprogramming due to inhibition of the master metabolic controller, c-Myc. Restoration of c-Myc by deletion of the bacterial asparaginase ansB reinvigorated T cell activation, but at the cost of decreased metabolic control of the tumour by STm. This work shows for the first time that T cells are metabolically defective during BCT, but also that this same phenomenon is inexorably tied to intrinsic tumour suppression by the bacterial vector. Synopsis: Attenuated Salmonella show promise as cancer therapeutics, yet T cells play no role in efficacy, limiting combined checkpoint blockade therapies or induction of immune memory. Improving T cell responses requires a better understanding of the causes of T cell dysfunction during Salmonella BCT. Attenuated Salmonella enterica Typhimurium (STm) cancer therapy drives T cell dysfunction. STm depletes asparagine via an asparaginase (ansB) in the tumour microenvironment (TME), suppressing tumour growth. Asparagine depletion in the TME triggers T cell metabolic arrest by destabilising c-Myc. An STm∆ansB mutant restores T cell function but at the cost of enhanced tumour metabolism. The STm∆ansB mutant still effectively reduces tumour growth in a mouse model. Attenuated Salmonella show promise as cancer therapeutics, yet T cells play no role in efficacy, limiting combined checkpoint blockade therapies or induction of immune memory. Improving T cell responses requires a better understanding of the causes of T cell dysfunction during Salmonella BCT. [ABSTRACT FROM AUTHOR]

OBJECTIVE To evaluate the pathologic ocular changes in a population of aquarium - housed Monodactylus argenteus and investigate potential underlying causes for the large number of affected fish in this exhibit. ANIMALS 11 Monodactylus argenteus were evaluated from a shoaling ring exhibit within an aquarium, and 19 control fish without ocular abnormalities were obtained from commercial fish suppliers. METHODS Physical and ocular examinations were performed antemortem. Postmortem samples of liver, heart, dorsal epaxial muscle, and lenses of affected and control fish were analyzed for amino acid profiles. The aqueous humor from affected and control fish was collected postmortem, and osmolality was analyzed. Tissues from affected and control fish were submitted for histopathology. RESULTS Ocular abnormalities in affected fish included corneal lesions, cataracts, lens capsule rupture, and unilateral left-sided lens luxation and buphthalmos. Lens luxation and buphthalmos were directly correlated. Aqueous humor osmolality in control fish differed significantly compared to affected fish but was not correlated to lens abnormality score. Affected fish had significantly lower lens concentrations of arginine, asparagine, glycine, isoleucine, serine, and tyrosine than control fish. One affected fish had severe buphthalmos, cataracts, and panophthalmitis caused by gram-positive cocci. CONCLUSIONS Cataracts and traumatic lesions were common in affected M argenteus. Dietary amino acid content and aqueous humor osmolality imbalances were not likely a cause of the cataracts. Differences in lens amino acid concentrations between affected and control fish may be related to cataract formation. CLINICAL RELEVANCE Consideration of species-appropriate tank parameters may mitigate ocular lesions including cataracts in aquarium fish. [ABSTRACT FROM AUTHOR]

Introduction: The Arabidopsis abc1k1/pgr6 (Activity of BC1 complex/proton regulation 6) mutant is characterized by photosynthetic and conditional developmental phenotypes triggered by stressful red as well as high light. The Arabidopsis ABC1-like kinases belong to the atypical kinase family and contain conserved ATP-binding and hydrolysis motifs, but their physiological requirement has never been investigated. Methods: By mutation to asparagine, we demonstrate that the highly conserved active site aspartate residue within ATP-binding motif VIIb is required for the physiological functions of ABC1K1. Results: Complementation of the abc1k1 knock out mutant with ABC1K1 D400N, failed to restore the wildtype phenotype. Discussion: These results provide in vivo evidence for a critical role of the active site aspartate residue (D400) of ABC1K1. [ABSTRACT FROM AUTHOR]

Human coronavirus (CoV) HKU1 infection typically causes common cold but can lead to pneumonia in children, older people, and immunosuppressed individuals. Recently, human transmembrane serine protease 2 (hTMPRSS2) was identified as the functional receptor for HKU1, but its region and residues critical for HKU1 S binding remain elusive. In this study, we find that HKU1 could utilize human and hamster, but not rat, mouse, or bat TMPRSS2 for virus entry, displaying a narrow host range. Using human-bat TMPRSS2 chimeras, we show that the serine peptidase (SP) domain of TMPRSS2 is essential for entry of HKU1. Further extensive mutagenesis analyses of the C-terminal regions of SP domains of human and bat TMPRSS2s identify residues 417 and 469 critical for entry of HKU1. Replacement of either D417 or Y469 with asparagine in hTMPRSS2 abolishes its abilities to mediate entry of HKU1 S pseudovirions and cell-cell fusion, whereas substitution of N417 with D or N469 with Y in bat TMPRSS2 (bTMPRSS2) renders it supporting HKU1 entry. Our findings contribute to a deeper understanding of coronavirus-receptor interactions and cross-species transmission. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Chinese Institute of Food Science & Technology / Zhongguo Shipin Xuebao is the property of Journal of Chinese Institute of Food Science & Technology Periodical Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

A transverse relaxation optimized spectroscopy (TROSY) approach is described for the optimal detection of NH2 groups in asparagine and glutamine side chains of proteins. Specifically, we have developed NMR experiments for isolating the slow-relaxing 15N and 1H components of NH2 multiplets. Although even modest sensitivity gains in 2D NH2-TROSY correlation maps compared to their decoupled NH2–HSQC counterparts can be achieved only occasionally, substantial improvements in resolution of the NMR spectra are demonstrated for asparagine and glutamine NH2 sites of a buried cavity mutant, L99A, of T4 lysozyme at 5 ºC. The NH2-TROSY approach is applied to CPMG relaxation dispersion measurements at the side chain NH2 positions of the L99A T4 lysozyme mutant — a model system for studies of the role of protein dynamics in ligand binding. [ABSTRACT FROM AUTHOR]

Abstract Background Intrauterine growth restriction (IUGR) affects up to 30% of piglets in a litter. Piglets exposed to IUGR prioritize brain development during gestation, resulting in a higher brain-to-liver weight ratio (BrW/LW) at birth. IUGR is associated with increased mortality, compromised metabolism, and gut health. However, the dynamic metabolic and microbial shifts in IUGR-affected pigs remain poorly understood. This study aimed to investigate the longitudinal effects of IUGR, defined by a high BrW/LW, on the composition of faecal microbiota and plasma metabolome in pigs from birth to slaughter. One day (± 1) after birth, computed tomography was performed on each piglet to assess their brain and liver weights. The pigs with the highest (IUGR = 12) and the lowest (NORM = 12) BrW/LW were selected to collect faeces and blood during lactation (day 16 ± 0.6, T1) and at the end of the starter period (day 63 ± 8.6, T2) and faeces at the beginning (day 119 ± 11.4, T3) and end of the finisher period (day 162 ± 14.3, T4). Results Faecal microbial Alpha diversity remained unaffected by IUGR across all time points. However, the Beta diversity was influenced by IUGR at T1 (P = 0.002), T2 (P = 0.08), and T3 (P = 0.03). Specifically, IUGR pigs displayed higher abundances of Clostridium sensu stricto 1 (P adj = 0.03) and Romboutsia (P adj = 0.05) at T1, Prevotellaceae NK3B31 group (P adj = 0.02), Rikenellaceae RC9 gut group (P adj = 0.03), and Alloprevotella (P adj = 0.03) at T2, and p-2534-18B5 gut group (P adj = 0.03) at T3. Conversely, the NORM group exhibited higher abundances of Ruminococcus (P adj = 0.01) at T1, HT002 (P adj = 0.05) at T2, and Prevotella_9 (P adj

This study established a novel method for measuring the contents of asparagine (Asn) and glutamine (Gln) in fruits and vegetables based on enzymatic hydrolysis combine with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). After enzymatic hydrolysis with Pronase E, samples were treated with butylated hydroxyanisole as an antioxidant. The supernatant was collected for analysis by UPLC-MS/MS. The chromatographic separation was accomplished on a Phenomenex kinetex F5 column using gradient elution with a mobile phase consisting of methanol and 0.02% formic acid in water. The mass spectrometer was equipped with an electrospray ionization source and operated in the positive ion mode with multiple reaction monitoring (MRM), and quantification was achieved using stable isotope-labeled internal standards. The calibration curves for Asn and Gln showed a good linearity in the range of 10–1 000 ng/mL (R2 > 0.999), with limit of detection (LOD) and limit of quantification (LOQ) of 1.5 and 5.0 mg/100 g, respectively. The recovery rates were in the range of 80.0%–112.8% with relative standard deviations (RSDs) of 1.2%–9.1%. The contents of Asn and Gln in 49 common vegetables and 50 common fruits were determined by the UPLC-MS/MS method. In vegetables, the average total content of Asn and Gln decreased in the order of bean > potato > onion and garlic > edible fungi > melon > solanaceous > cabbage. Notably, the total contents of Asn and Gln in beans were significantly higher than those in green leafy vegetables (P melons > aggregate fruits > berries > citrus > banana > nuts, and drupes had significantly higher total contents of Asn and Gln than nuts (P < 0.05). These findings suggested that individuals with urea cycle disorders should eat more nut fruits and green leafy vegetables, with less asparagine and glutamine, in daily life, especially vegetables and fruits with higher ratio of branched-chain amino acids to Asn plus Gln such as Chinese cabbage, Auricularia auricula, green onion, coconut juice, longan, and pomelo, and should eat less vegetables and fruits with lower ratio of branched-chain amino acids to Asn plus Gln such as potato, pumpkin, green bean, cantaloupe, red plum and yellow peach.

Malaria poses an enormous threat to human health. With ever increasing resistance to currently deployed drugs, breakthrough compounds with novel mechanisms of action are urgently needed. Here, we explore pyrimidine-based sulfonamides as a new low molecular weight inhibitor class with drug-like physical parameters and a synthetically accessible scaffold. We show that the exemplar, OSM-S-106, has potent activity against parasite cultures, low mammalian cell toxicity and low propensity for resistance development. In vitro evolution of resistance using a slow ramp-up approach pointed to the Plasmodium falciparum cytoplasmic asparaginyl-tRNA synthetase (PfAsnRS) as the target, consistent with our finding that OSM-S-106 inhibits protein translation and activates the amino acid starvation response. Targeted mass spectrometry confirms that OSM-S-106 is a pro-inhibitor and that inhibition of PfAsnRS occurs via enzyme-mediated production of an Asn-OSM-S-106 adduct. Human AsnRS is much less susceptible to this reaction hijacking mechanism. X-ray crystallographic studies of human AsnRS in complex with inhibitor adducts and docking of pro-inhibitors into a model of Asn-tRNA-bound PfAsnRS provide insights into the structure-activity relationship and the selectivity mechanism.

Abstract Bacterial cancer therapy (BCT) is a promising therapeutic for solid tumours. Salmonella enterica Typhimurium (STm) is well-studied amongst bacterial vectors due to advantages in genetic modification and metabolic adaptation. A longstanding paradox is the redundancy of T cells for treatment efficacy; instead, STm BCT depends on innate phagocytes for tumour control. Here, we used distal T cell receptor (TCR) and IFNγ reporter mice (Nr4a3-Tocky-Ifnγ-YFP) and a colorectal cancer (CRC) model to interrogate T cell activity during BCT with attenuated STm. We found that colonic tumour infiltrating lymphocytes (TILs) exhibited a variety of activation defects, including IFN-γ production decoupled from TCR signalling, decreased polyfunctionality and reduced central memory (TCM) formation. Modelling of T-cell–tumour interactions with a tumour organoid platform revealed an intact TCR signalosome, but paralysed metabolic reprogramming due to inhibition of the master metabolic controller, c-Myc. Restoration of c-Myc by deletion of the bacterial asparaginase ansB reinvigorated T cell activation, but at the cost of decreased metabolic control of the tumour by STm. This work shows for the first time that T cells are metabolically defective during BCT, but also that this same phenomenon is inexorably tied to intrinsic tumour suppression by the bacterial vector.

The age‐associated decline in intestinal stem cell (ISC) function is a key factor in intestinal aging in organisms, resulting in impaired intestinal function and increased susceptibility to age‐related diseases. Consequently, it is imperative to develop effective therapeutic strategies to prevent ISC aging and functional decline. In this study, we utilized an aging Drosophila model screening of amino acids and found that asparagine (Asn), a nonessential amino acid in vivo, exhibits its profound anti‐aging properties on ISCs. Asn inhibits the hyperproliferation of aging ISCs in Drosophila, maintains intestinal homeostasis, and extends the lifespan of aging flies. Complementarily, Asn promotes the growth and branching of elderly murine intestinal organoids, indicating its anti‐aging capacity to enhance ISC function. Mechanistic analyses have revealed that Asn exerts its effects via the activation of the autophagic signaling pathway. In summary, this study has preliminarily explored the potential supportive role of Asn in ameliorating intestinal aging, providing a foundation for further research into therapeutic interventions targeting age‐related intestinal dysfunction. [ABSTRACT FROM AUTHOR]

BACKGROUND RESULTS CONCLUSION Low‐temperature sweetening is a crucial aspect in the potato chips industry because of the initiation of browning. The present study aimed to investigate the effect of ultrasonication on the concentrations of reducing sugars, sucrose, asparagine and chip color on tubers with different characteristics. Therefore, samples of the cultivars (cvs.) Nicola, Laura and Golden Wonder were used in this study. The tubers were initially stored at 4 °C and a relative humidity of 90% for 18 weeks. Ultrasonication was applied at 25 and 50 °C for 10, 20 and 30 min.The results showed that ultrasonication contributed to improve the color of the chips produced from the chip cultivar (cv. Golden Wonder), as well as the non‐chip cultivars (cvs. Nicola and Laura). The concentration of reducing sugars and sucrose is the main factor for the color development of the chips. Increased temperature and extended ultrasonication duration correlate with more pronounced cell wall disruption.The present study revealed that ultrasonication could be considered as a pretreatment for production of chips from cold‐stored tubers. However, it is necessary to validate the feasibility of this application in an upscaled process. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Among the 20 proteinogenic amino acids, glutamine (GLN) and asparagine (ASN) represent a unique cohort in containing a terminal amide in their side chain, and share a direct metabolic relationship, with glutamine generating asparagine through the ATP-dependent asparagine synthetase (ASNS) reaction. Circulating glutamine levels and metabolic flux through cells and tissues greatly exceed those for asparagine, and "glutamine addiction" in cancer has likewise received considerable attention. However, historic and recent evidence collectively suggest that in spite of its modest presence, asparagine plays an outsized regulatory role in cellular function. Here, we present a unifying evidence-based hypothesis that the amides constitute a regulatory signaling circuit, with glutamine as a driver and asparagine as a second messenger that allosterically regulates key biochemical and physiological functions, particularly cell growth and survival. Specifically, it is proposed that ASNS serves as a sensor of substrate sufficiency for S-phase entry and progression in proliferating cells. ASNS-generated asparagine serves as a subsequent second messenger that modulates the activity of key regulatory proteins and promotes survival in the face of cellular stress, and serves as a feed-forward driver of S-phase progression in cell growth. We propose that this signaling pathway be termed the amide signaling circuit (ASC) in homage to the SLC1A5-encoded ASCT2 that transports both glutamine and asparagine in a bidirectional manner, and has been implicated in the pathogenesis of a broad spectrum of human cancers. Support for the ASC model is provided by the recent discovery that glutamine is sensed in primary cilia via ASNS during metabolic stress. [ABSTRACT FROM AUTHOR]

Background: The intricate interplay between peripheral adaptive immune cells and the central nervous system (CNS) has garnered increasing recognition. Given that alterations in cell quantities often translate into modifications in metabolite profiles and that these metabolic changes can potentially traverse the bloodstream and enter the CNS, thereby modulating the progression of mental illnesses, we sought to explore the metabolic profiles of peripheral immune cells in a ketamine-treated mouse model of schizophrenia. Methods: We used flow cytometry to scrutinize the alterations in peripheral adaptive immune cells in a ketamine-induced schizophrenia mouse model. Subsequently, we implemented an untargeted metabolomic approach with ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) to detect the metabolite profiles of peripheral abnormal lymphocytes and identify differential metabolites present in plasma. We then employed targeted metabolomics using UPLC-MS/MS to quantify the common differential metabolites detected in mouse plasma. Results: Flow cytometry analysis detected a notable increase in the count of peripheral CD3+ T cells in a ketamine-induced schizophrenia mouse model. Subsequent untargeted metabolomics analysis revealed that the amino acid metabolism pathway underwent substantial alterations. A detailed quantification of 22 amino acid profiles in the peripheral plasma indicated significant elevation in the levels of glycine, alanine, asparagine, and aspartic acid. Limitations: Our ongoing research has yet to conclusively identify the precise amino acid metabolism pathway that serves as the pivotal factor in the manifestation of the schizophrenia-like phenotype induced by ketamine. Conclusion: The peripheral amino acid metabolism pathway is involved in the ketamine-induced schizophrenia-like phenotype. The metabolic profile of peripheral immune cells could provide accurate biomarkers for the diagnosis and treatment of psychiatric diseases. [ABSTRACT FROM AUTHOR]

Infectious spleen and kidney necrosis virus (ISKNV) has brought huge economic loss to the aquaculture industry. Through interfering with the viral replication and proliferation process that depends on host cells, its pathogenicity can be effectively reduced. In this study, we investigated the role of asparagine metabolites in ISKNV proliferation. The results showed that ISKNV infection up-regulated the expression of some key enzymes of the asparagine metabolic pathway in Chinese perch brain (CPB) cells. These key enzymes, including glutamic oxaloacetic transaminase 1/2 (GOT1/2) and malate dehydrogenase1/2 (MDH1/2) associated with the malate-aspartate shuttle (MAS) pathway and asparagine synthetase (ASNS) involved in the asparagine biosynthesis pathway, were up-regulated during ISKNV replication and release stages. In addition, results showed that the production of ISKNV was significantly reduced by inhibiting the MAS pathway or reducing the expression of ASNS by 1.3-fold and 0.6-fold, respectively, indicating that asparagine was a critical limiting metabolite for ISKNV protein synthesis. Furthermore, when asparagine was added to the medium without glutamine, ISKNV copy number was restored to 92% of that in the complete medium, indicating that ISKNV could be fully rescued from the absence of glutamine by supplementing asparagine. The above results indicated that asparagine was a critical factor in limiting the effective replication of ISKNV, which provided a new idea for the treatment of aquatic viral diseases. [ABSTRACT FROM AUTHOR]

• Amaranthus caudatus and Amaranthus cruentus are common amaranth species consumed in South Africa. • Pre-harvest application of different concentrations of phytostim biostimulant were investigated. • Growth and yield components were improved by the application of 1.5 % concentration in both studied species. • Application of 0.5 and 1.0 % improved the protein content, some minerals and bioactive components. The current study aims to evaluate the effect of pre-harvest application of different concentrations of phytostim biostimulant on growth, yield, nutrition, phenolic acids and some phenolic compound in two species of Amaranthus. A two factor greenhouse trial was established in a split-plot to accommodate eight phytostim biostimulant concentrations (0, 0.5, 1, 1.5, 2.5, 3, 4.5, and 6%) in Amaranthus cruentus and Amaranthus caudatus for up to 60 days post-transplant. Application of 1.5% phytostim biostimulant resulted in the highest growth and biomass accumulation, whilst concentrations of >3% resulted in reduced growth and biomass yield in both studied species. These results were in concomitant to an enhanced accumulation of total phenols, total flavonoids, β-carotene, and antioxidants activity (DPPH and ABTS) in Amaranthus cruentus. Application of 1% in Amaranthus cruentus had a positive effect of the accumulation of zinc, iron and calcium and some amino acids (alanine, asparagine, lysine and arginine). In Amaranthus caudatus , 0.5% concentration resulted in the highest total phenols, total flavonoids, antioxidant activity (DPPH and ABTS), chlorophyll b and total chlorophyll. Amaranthus cruentus 0.5% resulted in the highest 2-O-caffeoylglucaric acid, whilst application of 1.0% in Amaranthus caudatus showed the highest caffeic acid, 3-O-glucuronide thus suggesting. Thus, suggesting that phytostim biostimulant can be used to manipulate growth and nutritional components in the studied Amaranthus species. Lower concentrations (0.5–1.5%) of phytostim biostimulant can be recommended for cultivation of A. Cr. A. cruentus and Amaranthus caudatus. [ABSTRACT FROM AUTHOR]

SIRT5, one of the mammalian sirtuins, specifically recognizes succinyl‐lysine residues on proteins and catalyzes the desuccinylation reaction. In this study, we characterized SIRT5 mutants with hydrophobic amino acid substitutions at Q140 and N141, in addition to the catalytic residue H158, known as an active site residue, by the Michaelis–Menten analysis and X‐ray crystallography. Kinetic analysis showed that the catalytic efficiency (kcat/Km) of the Q140L and N141V mutants decreased to 0.02 times and 0.0038 times that of the wild‐type SIRT5, respectively, with the activity of the N141V mutant becoming comparable to that of the H158M mutant. Our findings indicate that N141 contributes significantly to the desuccinylation reaction. [ABSTRACT FROM AUTHOR]

ABCA4 is an ATP-binding cassette (ABC) transporter that prevents the buildup of toxic retinoid compounds by facilitating the transport of N-retinylidene-phosphatidylethanolamine across membranes of rod and cone photoreceptor cells. Over 1500 missense mutations in ABCA4, many in the nucleotide-binding domains (NBDs), have been genetically linked to Stargardt disease. Here, we show by cryo-EM that ABCA4 is converted from an open outward conformation to a closed conformation upon the binding of adenylylimidodiphosphate. Structural information and biochemical studies were used to further define the role of the NBDs in the functional properties of ABCA4 and the mechanisms by which mutations lead to the loss in activity. We show that ATPase activity in both NBDs is required for the functional activity of ABCA4. Mutations in Walker A asparagine residues cause a severe reduction in substrate-activated ATPase activity due to the loss in polar interactions with residues within the D-loops of the opposing NBD. The structural basis for how disease mutations in other NBD residues, including the R1108C, R2077W, R2107H, and L2027F, affect the structure and function of ABCA4 is described. Collectively, our studies provide insight into the structure and function of ABCA4 and mechanisms underlying Stargardt disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Liposarcoma is a rare cancer of adipose tissue with limited treatment options. Here, we studied the fuel used by liposarcoma to develop a new strategy for treatment. Liposarcoma was found to rely on the amino acid Asparagine for tumor growth, especially in its most aggressive form. By combining treatments that limit both synthesis and uptake of Asparagine within liposarcoma cells, we demonstrated a unique sensitivity that reduced tumor growth in animal models. Altogether, findings from this study suggest that targeting Asparagine could be a promising new therapy in liposarcoma. Background: mTORC1 activity is dependent on the presence of micronutrients, including Asparagine (Asn), to promote anabolic cell signaling in many cancers. We hypothesized that targeting Asn metabolism would inhibit tumor growth by reducing mTORC1 activity in well-differentiated (WD)/dedifferentiated (DD) liposarcoma (LPS). Methods: Human tumor metabolomic analysis was utilized to compare abundance of Asn in WD vs. DD LPS. Gene set enrichment analysis (GSEA) compared relative expression among metabolic pathways upregulated in DD vs. WD LPS. Proliferation assays were performed for LPS cell lines and organoid models by using the combination treatment of electron transport chain (ETC) inhibitors with Asn-free media. 13C-Glucose-labeling metabolomics evaluated the effects of combination treatment on nucleotide synthesis. Murine xenograft models were used to assess the effects of ETC inhibition combined with PEGylated L-Asparaginase (PEG-Asnase) on tumor growth and mTORC1 signaling. Results: Asn was enriched in DD LPS compared to WD LPS. GSEA indicated that mTORC1 signaling was upregulated in DD LPS. Within available LPS cell lines and organoid models, the combination of ETC inhibition with Asn-free media resulted in reduced cell proliferation. Combination treatment inhibited nucleotide synthesis and promoted cell cycle arrest. In vivo, the combination of ETC inhibition with PEG-Asnase restricted tumor growth. Conclusions: Asn enrichment and mTORC1 upregulation are important factors contributing to WD/DD LPS tumor progression. Effective targeting strategies require limiting access to extracellular Asn and inhibition of de novo synthesis mechanisms. The combination of PEG-Asnase with ETC inhibition is an effective therapy to restrict tumor growth in WD/DD LPS. [ABSTRACT FROM AUTHOR]

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Colorectal cancer (CRC) is a multi-stage process initiated through the formation of a benign adenoma, progressing to an invasive carcinoma and finally metastatic spread. Tumour cells must adapt their metabolism to support the energetic and biosynthetic demands associated with disease progression. As such, targeting cancer cell metabolism is a promising therapeutic avenue in CRC. However, to identify tractable nodes of metabolic vulnerability specific to CRC stage, we must understand how metabolism changes during CRC development. Here, we use a unique model system – comprising human early adenoma to late adenocarcinoma. We show that adenoma cells transition to elevated glycolysis at the early stages of tumour progression but maintain oxidative metabolism. Progressed adenocarcinoma cells rely more on glutamine-derived carbon to fuel the TCA cycle, whereas glycolysis and TCA cycle activity remain tightly coupled in early adenoma cells. Adenocarcinoma cells are more flexible with respect to fuel source, enabling them to proliferate in nutrient-poor environments. Despite this plasticity, we identify asparagine (ASN) synthesis as a node of metabolic vulnerability in late-stage adenocarcinoma cells. We show that loss of asparagine synthetase (ASNS) blocks their proliferation, whereas early adenoma cells are largely resistant to ASN deprivation. Mechanistically, we show that late-stage adenocarcinoma cells are dependent on ASNS to support mTORC1 signalling and maximal glycolytic and oxidative capacity. Resistance to ASNS loss in early adenoma cells is likely due to a feedback loop, absent in late-stage cells, allowing them to sense and regulate ASN levels and supplement ASN by autophagy. Together, our study defines metabolic changes during CRC development and highlights ASN synthesis as a targetable metabolic vulnerability in later stage disease.

Abstract Background Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease without meaningful therapeutic options beyond the first salvage therapy. Targeting PDAC metabolism through amino acid restriction has emerged as a promising new strategy, with asparaginases, enzymes that deplete plasma glutamine and asparagine, reaching clinical trials. In this study, we investigated the anti-PDAC activity of the asparaginase formulation Pegcrisantaspase (PegC) alone and in combination with standard-of-care chemotherapeutics. Methods Using mouse and human PDAC cell lines, we assessed the impact of PegC on cell proliferation, cell death, and cell cycle progression. We further characterized the in vitro effect of PegC on protein synthesis as well as the generation of reactive oxygen species and levels of glutathione, a major cellular antioxidant. Additional cell line studies examined the effect of the combination of PegC with standard-of-care chemotherapeutics. In vivo, the tolerability and efficacy of PegC, as well as the impact on plasma amino acid levels, was assessed using the C57BL/6-derived KPC syngeneic mouse model. Results Here we report that PegC demonstrated potent anti-proliferative activity in a panel of human and murine PDAC cell lines. This decrease in proliferation was accompanied by inhibited protein synthesis and decreased levels of glutathione. In vivo, PegC was tolerable and effectively reduced plasma levels of glutamine and asparagine, leading to a statistically significant inhibition of tumor growth in a syngeneic mouse model of PDAC. There was no observable in vitro or in vivo benefit to combining PegC with standard-of-care chemotherapeutics, including oxaliplatin, irinotecan, 5-fluorouracil, paclitaxel, and gemcitabine. Notably, PegC treatment increased tumor expression of asparagine and serine biosynthetic enzymes. Conclusions Taken together, our results demonstrate the potential therapeutic use of PegC in PDAC and highlight the importance of identifying candidates for combination regimens that could improve cytotoxicity and/or reduce the induction of resistance pathways.

Acute hepatic injury is observed in response to various stressors, including trauma, ingestion of hepatic toxins, and hepatitis. Investigations to date have focused on extrinsic and intrinsic signals required for hepatocytes to proliferate and regenerate the liver in response to injury, though there is a more limited understanding of induced stress responses promoting hepatocyte survival upon acute injury. In this issue of the JCI, Sun and colleagues detail a mechanism by which local activation of the nuclear receptor liver receptor homolog-1 (LRH-1; NR5A2) directly induces de novo asparagine synthesis and expression of asparagine synthetase (ASNS) in response to injury and show that this response restrains hepatic damage. This work opens up several avenues for inquiry, including the potential for asparagine supplementation to ameliorate acute hepatic injury.

Peste des petits ruminants (PPR), a contagious virus that infects sheep and goats, damages livestock globally. This study examined the clinical features and phylogenetic analysis of the PPR virus in Iraqi breed sheep from Al‐Diwaniyah province. A clinical trial of 610 sheep from different flocks found 150 oral lesions. Special primers for RT‐PCR and Mega11 for phylogenetic analysis were used to study the PPR virus nucleoprotein (N) gene. The PPR infection rate was 44.6% in 4–12 month olds (n = 33/131) and 4.8% in 36–48 month olds (n = 3/75). A 608‐bp PPR virus partial N gene sequence was found in 49.3% of samples by RT‐PCR. In leucine, isoleucine, proline, glycine, alanine, glutamine, asparagine, threonine, serine, arginine, and lysine codons, 25 amino acid alterations were found. The protein codon 56 alanine‐valine alteration was most significant. Moving from a smaller hydrophobic amino acid to one with a bigger side chain may reduce protein stability. Steric hindrance or protein shape change from Valine's extended side chain may impact folding, stability, functionality, and interactions with other molecules. Furthermore, phylogenetic analysis showed that the Nigerian strain (MN271586) was most similar to our Iraqi strain, with 100% identity and coverage. This study found the Peste des Petits Ruminants (PPR) virus in sheep flocks in Al‐Diwaniyah Governorate, Iraq, which is genetically similar to neighboring countries. PPR virus strains must be monitored and genetically characterized since N gene alterations can affect infection and propagation. [ABSTRACT FROM AUTHOR]

Photorespiration, caused by oxygenation of the enzyme Rubisco, is considered a wasteful process, because it reduces photosynthetic carbon gain, but it also supplies amino acids and is involved in amelioration of stress. Here, we show that a sudden increase in photorespiratory activity not only reduced carbon acquisition and production of sugars and starch, but also affected diurnal dynamics of amino acids not obviously involved in the process. Flux calculations based on diurnal metabolite profiles suggest that export of proline from leaves increases, while aspartate family members accumulate. An immense increase is observed for turnover in the cyclic reaction of glutamine synthetase/glutamine-oxoglutarate aminotransferase (GS/GOGAT), probably because of increased production of ammonium in photorespiration. The hpr1-1 mutant, defective in peroxisomal hydroxypyruvate reductase, shows substantial alterations in flux, leading to a shift from the oxoglutarate to the aspartate family of amino acids. This is coupled to a massive export of asparagine, which may serve in exchange for serine between shoot and root. [ABSTRACT FROM AUTHOR]

The amide proteogenic amino acids, asparagine and glutamine, are two of the twenty amino acids used in translation by all known life. The aminoacyl‐tRNA synthetases for asparagine and glutamine, asparaginyl‐tRNA synthetase and glutaminyl tRNA synthetase, evolved after the split in the last universal common ancestor of modern organisms. Before that split, life used two‐step indirect pathways to synthesize asparagine and glutamine on their cognate tRNAs to form the aminoacyl‐tRNA used in translation. These two‐step pathways were retained throughout much of the bacterial and archaeal domains of life and eukaryotic organelles. The indirect routes use non‐discriminating aminoacyl‐tRNA synthetases (non‐discriminating aspartyl‐tRNA synthetase and non‐discriminating glutamyl‐tRNA synthetase) to misaminoacylate the tRNA. The misaminoacylated tRNA formed is then transamidated into the amide aminoacyl‐tRNA used in protein synthesis by tRNA‐dependent amidotransferases (GatCAB and GatDE). The enzymes and tRNAs involved assemble into complexes known as transamidosomes to help maintain translational fidelity. These pathways have evolved to meet the varied cellular needs across a diverse set of organisms, leading to significant variation. In certain bacteria, the indirect pathways may provide a means to adapt to cellular stress by reducing the fidelity of protein synthesis. The retention of these indirect pathways versus acquisition of asparaginyl‐tRNA synthetase and glutaminyl tRNA synthetase in lineages likely involves a complex interplay of the competing uses of glutamine and asparagine beyond translation, energetic costs, co‐evolution between enzymes and tRNA, and involvement in stress response that await further investigation. [ABSTRACT FROM AUTHOR]

The mechanism by which nitrate inhibits nitrogen fixation in soybean (Glycine max L.) is not fully understood. Accumulation of ureide in soybean plant tissues may regulate the nitrogen fixation capacity through a feedback pathway. In this study, unilaterally nodulated dual-root soybeans prepared by grafting were grown in sand culture. They were subjected to the removal of the nodulated side roots, and were given either nitrate supply or no supply to the non-nodulated side roots for 3 days (experiment I). Additionally, they received nitrate supply to the non-nodulated side roots for 1–14 days (experiment II). The results showed that nitrate supply increased the levels of asparagine and ureide in soybean shoots (Experiment I). In Experiment II, nodule dry weight, nodule number, nodule nitrogenase activity, and nodule urate oxidase activity decreased significantly after 3, 7, and 14 days of nitrate supply. Ureide content in the shoots and nodules increased after 1, 3, and 7 days of nitrate supply, but decreased after 14 days of nitrate supply. There was a significant positive correlation between urate oxidase activity and nitrogenase activity. Hence, we deduced that nitrate supply increased the asparagine content in soybean shoots, likely inhibiting ureide degradation, which induced the accumulation of ureide in soybean shoots and nodules, and, in turn, feedback inhibited the nodule nitrogen fixation. In addition, urate oxidase activity can be used to assess the nitrogen fixation capacity of nodules. [ABSTRACT FROM AUTHOR]

The ninhydrin reaction is commonly used for the detection of amino acids. However, in the literature, different conditions with respect to the buffer system, its pH and concentration, type of organic solvent, incubation time, and temperature, as well as the concentrations of the reagents, are described. To identify the most suitable conditions, colour development with reagents of varying compositions and different reaction temperatures and times were investigated using asparagine as a model amino acid. Asparagine was selected since it is one of the most abundant free amino acids in many types of samples. The optimal reaction mixture consisted of 0.8 mol L−1 potassium acetate, 1.6 mol L−1 acetic acid, 20 mg mL−1 ninhydrin and 0.8 mg mL−1 hydrindantin in DMSO/acetate buffer 40/60 (v/v) (final concentrations). The best reaction condition was heating the samples in 1.5 mL reaction tubes to 90 °C for 45 min. Afterwards, the samples were diluted with 2-propanol/water 50/50 (v/v) and the absorbance was measured at 570 nm. The proteinogenic amino acids showed a similar response except for cysteine and proline. The method was highly sensitive and showed excellent linearity as well as intra-day and inter-day reproducibility. [ABSTRACT FROM AUTHOR]

Site-specific modifications of aspartate residues spontaneously occur in crystallin, the major protein in the lens. One of the primary modification sites is Asp151 in αA-crystallin. Isomerization and racemization alter the crystallin backbone structure, reducing its stability by inducing abnormal crystallin–crystallin interactions and ultimately leading to the insolubilization of crystallin complexes. These changes are considered significant factors in the formation of senile cataracts. However, the mechanisms driving spontaneous isomerization and racemization have not been experimentally demonstrated. In this study, we generated αA-crystallins with different homo-oligomeric sizes and/or containing an asparagine residue at position 151, which is more prone to isomerization and racemization. We characterized their structure, hydrophobicity, chaperone-like function, and heat stability, and examined their propensity for isomerization and racemization. The results show that the two differently sized αA-crystallin variants possessed similar secondary structures but exhibited different chaperone-like functions depending on their oligomeric sizes. The rate of isomerization and racemization of Asp151, as assessed by the deamidation of Asn151, was also found to depend on the oligomeric sizes of αA-crystallin. The predominant isomerization product via deamidation of Asn151 in the different-sized αA-crystallin variants was L-β-Asp in vitro, while various modifications occurred around Asp151 in vivo. The disparity between the findings of this in vitro study and in vivo studies suggests that the isomerization of Asp151 in vivo may be more complex than what occurs in vitro. [ABSTRACT FROM AUTHOR]

Summary: B‐cell precursor acute lymphoblastic leukaemia (BCP‐ALL) blasts strictly depend on the transport of extra‐cellular asparagine (Asn), yielding a rationale for L‐asparaginase (ASNase) therapy. However, the carriers used by ALL blasts for Asn transport have not been identified yet. Exploiting RS4;11 cells as BCP‐ALL model, we have found that cell Asn is lowered by either silencing or inhibition of the transporters ASCT2 or SNAT5. The inhibitors V‐9302 (for ASCT2) and GluγHA (for SNAT5) markedly lower cell proliferation and, when used together, suppress mTOR activity, induce autophagy and cause a severe nutritional stress, leading to a proliferative arrest and a massive cell death in both the ASNase‐sensitive RS4;11 cells and the relatively ASNase‐insensitive NALM‐6 cells. The cytotoxic effect is not prevented by coculturing leukaemic cells with primary mesenchymal stromal cells. Leukaemic blasts of paediatric ALL patients express ASCT2 and SNAT5 at diagnosis and undergo marked cytotoxicity when exposed to the inhibitors. ASCT2 expression is positively correlated with the minimal residual disease at the end of the induction therapy. In conclusion, ASCT2 and SNAT5 are the carriers exploited by ALL cells to transport Asn, and ASCT2 expression is associated with a lower therapeutic response. ASCT2 may thus represent a novel therapeutic target in BCP‐ALL. [ABSTRACT FROM AUTHOR]

Simple Summary: Due to the growing world population, the sustainability of food and feed sources with high nutritional value has become a crucial issue. In this scenario, insects can constitute a low-impact protein source with high nutritional value. The growth and nutrient composition of insects are potentially influenced by rearing conditions, particularly by the selected growth substrate. In this study, we evaluated the chemical and functional characteristics of Tenebrio molitor larvae reared on different growth substrates: a traditional wheat bran substrate and an innovative substrate consisting of wheat bran supplemented with chestnut shell, a by-product of the chestnut agro-industrial chain. The results showed that the innovative growth substrates positively influenced the insects' survival suggesting a beneficial effect on larval health. The enrichment of the growth substrate with chestnut shell modified the protein and amino acid profile of insect meals, possibly indicating a shift in their metabolism. In addition, insect meals obtained from larvae reared on chestnut-shell-enriched substrate exhibited higher antibacterial and antioxidant activity, suggesting a potential beneficial effect when included in animal feed. Our results showed positive outcomes related to the design of innovative strategies for insect rearing, enriching larvae meal with beneficial health properties in line with sustainability and One Health principles. Tenebrio molitor larvae represent a sustainable protein source for food and feed. The aim of this study was to evaluate the supplementation of chestnut shell, a by-product of the agro-industrial chain, in growth substrates for T. molitor larvae rearing. Seven-week-old larvae were reared on three different growth substrates: the control group (CTRL) was fed wheat bran, treatment group one was fed wheat bran supplemented with 12.5% w/w chestnut shell (TRT1), and treatment group two was fed wheat bran supplemented with 25% w/w chestnut shell (TRT2). Larval weight, substrate consumption, and mortality were recorded weekly. After 14 days, insect meals were produced for bromatological and colorimetric analysis, and bacterial inhibition activity assay using a microdilution method. The amino acid profile of insects was determined using quantitative nuclear magnetic resonance spectroscopy. Our results showed a lower feed conversion ratio and higher larval survival rate % in TRT2 compared to CTRL (p < 0.05). Proteins and lipids of TRT2 were higher than other groups (p < 0.05). Important differences were observed in the amino acid profile of TRT1 and TRT2 compared to CTRL (p < 0.05). TRT1 and TRT2 showed higher E. coli inhibitory activity than CTRL (p < 0.05). In conclusion, chestnut shell supplementation improved the survival and functional characteristics of larvae and likely impacted the insects' metabolism. [ABSTRACT FROM AUTHOR]

Introduction: This study evaluated nutrient deficiencies in infants and toddlers with inflammatory bowel disease (IBD) and eosinophilic gastrointestinal disorders (EGIDs), whose primary nutritional source is elemental formulas (EFs). Methods: The nutrient status of children with IBD and EGID aged 6 months to 6 years was evaluated. Results: Twenty-one children fed with EFs (EF group) and 25 controls (CL group) were enrolled. The selenium level in the EF group was lower than that in the CL group (2.2 μg/dL vs. 9.3 μg/dL; p < 0.01). Although fat-soluble vitamins were deficient in some EF group participants, no significant differences were observed in their concentration and insufficiency proportion. However, ascorbic acid deficiency was more frequent in the EF group, with significantly lower levels (8.6 μg/mL vs. 12.0 μg/mL; p < 0.01). The triene:tetraene ratio was significantly higher in the EF group (0.046 vs. 0.010; p < 0.01). Asparagine and taurine levels were significantly lower in the EF group (asparagine: p < 0.01; taurine: p < 0.01) and tyrosine and phenylalanine levels were higher in the EF group, resulting in a lower Fisher's ratio (p < 0.01). Conclusion: Long-term feeding with EFs can cause deficiencies in essential fatty acids, selenium, and ascorbic acid and also carries a risk of amino acid imbalance in infants and toddlers. [ABSTRACT FROM AUTHOR]

The complete quantitative essential amino acid (EAA) requirements of aquacultured animals are largely unknown except for a handful of species. This is problematic because formulation of least-cost diets for target animals demands precise knowledge of EAA requirements. One way of approximating EAA requirements is to use correlations between whole-body and/or muscle EAA profiles using essential to total EAA ratios (A/E ratios). This method requires a reference, quantified EAA requirement, usually lysine (Lys). To systematically evaluate the A/E ratio method, muscle AA profiles were measured in 10 species of teleost and whiteleg shrimp. In fish, Lys represented the dominant muscle EAA measured, ranging between 7.06 and 9.58 g/100 g protein, whereas, in shrimp, arginine (Arg) was quantified as the principal EAA. For non-EAA, glutamate (Glu) and glutamine were consistently the NEAA recorded at highest levels whereas, in shrimp, Glu + Gln was highest followed by aspartate (Asp) plus asparagine (Asn). Except for Arg, which was twice that of fish (P < 0.05), whiteleg shrimp expressed similar muscle A/E profiles. Strength of relationship between muscle EAA levels and known requirements were all strongly and positively correlated for non-salmonid species. When using Lys as the reference EAA there were few semblances between measured and estimated EAA requirements. [ABSTRACT FROM AUTHOR]

The amine group (NH2) and the carboxyl group (COOH-) are the fundamental components of amino acids. They regulate several essential physiological functions of organisms and are found in all forms of life, including humans, plants, and microorganisms. Furthermore, they serve as vital energy sources and also act as neurotransmitters. An analysis of the kidneys of Pterocles alchata, an Iraqi Pin-tailed sandgrouse, revealed the presence of 18 specific amino acids. The data indicate that there were no significant differences in the total amounts of amino acids among the three renal lobes. The concentrations in the anterior, middle, and posterior lobes were measured to be 14.154±97.273, 12.437±87.255, and 11.882±88.157 correspondingly, with a significance level of p≤0.05. There were considerable variations in the amounts of amino acids among different areas of the kidney. A glutamine, serine, and asparagine surplus was most pronounced in the kidney lobes. Glutamate proportions were 14.38±226.661 in the back lobe, 22.9±231.929 in the middle lobe, and 27.74±261.852 in the front lobe. The statistical significance of these changes was determined using a significance level of p≤0.05.. Applying a probability threshold of p≤0.01, the serine concentrations in the kidney lobes were measured as 29.59±226.65, 17.74±202.183, and 7.71±199.840 in the anterior, middle, and posterior lobes, respectively. The concentration of the amino acid asparagine (Asn) in the anterior lobe was 12.61±153.952, in the middle lobe it was 7.47±135.278, and in the posterior lobe it was 10.23±128.885. These concentrations were observed at a probability threshold of p≤0.05 [ABSTRACT FROM AUTHOR]

Background: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease without meaningful therapeutic options beyond the first salvage therapy. Targeting PDAC metabolism through amino acid restriction has emerged as a promising new strategy, with asparaginases, enzymes that deplete plasma glutamine and asparagine, reaching clinical trials. In this study, we investigated the anti-PDAC activity of the asparaginase formulation Pegcrisantaspase (PegC) alone and in combination with standard-of-care chemotherapeutics. Methods: Using mouse and human PDAC cell lines, we assessed the impact of PegC on cell proliferation, cell death, and cell cycle progression. We further characterized the in vitro effect of PegC on protein synthesis as well as the generation of reactive oxygen species and levels of glutathione, a major cellular antioxidant. Additional cell line studies examined the effect of the combination of PegC with standard-of-care chemotherapeutics. In vivo, the tolerability and efficacy of PegC, as well as the impact on plasma amino acid levels, was assessed using the C57BL/6-derived KPC syngeneic mouse model. Results: Here we report that PegC demonstrated potent anti-proliferative activity in a panel of human and murine PDAC cell lines. This decrease in proliferation was accompanied by inhibited protein synthesis and decreased levels of glutathione. In vivo, PegC was tolerable and effectively reduced plasma levels of glutamine and asparagine, leading to a statistically significant inhibition of tumor growth in a syngeneic mouse model of PDAC. There was no observable in vitro or in vivo benefit to combining PegC with standard-of-care chemotherapeutics, including oxaliplatin, irinotecan, 5-fluorouracil, paclitaxel, and gemcitabine. Notably, PegC treatment increased tumor expression of asparagine and serine biosynthetic enzymes. Conclusions: Taken together, our results demonstrate the potential therapeutic use of PegC in PDAC and highlight the importance of identifying candidates for combination regimens that could improve cytotoxicity and/or reduce the induction of resistance pathways. [ABSTRACT FROM AUTHOR]

Critical seagrass ecosystems are predicted to be impacted by droughts and floods of increasing severity and frequency as the climate changes. Hyposalinity events alter seagrass composition and challenge resilience, yet understanding of the response mechanisms is incomplete but necessary for predicting outcomes. Hyposalinity stress response mechanisms in Halodule wrightii (shoal grass), a seagrass with wide salinity tolerance, were examined in mesocosm experiments by measuring monovalent and divalent ions, free amino acid (FAA) concentrations, and the Thiobarbituric Acid Reactive Substances (TBARS) indicator of lipid peroxidation. Two progressive, 3-step treatments, to moderate and extreme hyposalinity endpoints S15 and S5 were compared. Concentrations of Na+, K+, Ca2+, and Cl− decreased two to threefold with greater hyposalinity. Proline, the most abundant FAA initially, progressively decreased with increasing hyposalinity as asparagine became dominant. Glutamine and serine increased significantly, and all four FAA exhibited consistent patterns of change. Results are consistent with proline's role as osmolyte and regulator, and suggest that asparagine mobilizes or stores N, possibly due to another limiting nutrient. TBARS changed transiently at intermediate salinities, but were not significantly different at hyposalinity endpoints indicating that H. wrightii may have already reached stress equilibrium. Prolonged hyposalinity events may have consequences for seagrass resilience. [ABSTRACT FROM AUTHOR]

This article discusses the discovery of two enzymes, SilE-R and SilE-S, that have the ability to hydrolyze silylethers. These enzymes belong to the Stress-Response-A/B-Barrel-Domain-containing Proteins (DABB) family and exhibit enantiospecific hydrolysis of silylethers. They accept a wide range of substrates, including TBDMS-protected alcohols. The natural function of SilE-R and SilE-S is still unknown, but they have potential applications in synthetic chemistry. The article also explores the structure and mechanism of these enzymes. The study was funded by the German Research Foundation and the Engineering and Physical Sciences Research Council. [Extracted from the article]

The eukaryotic asparagine (N)-linked glycan is pre-assembled as a fourteen-sugar oligosaccharide on a lipid carrier in the endoplasmic reticulum (ER). Seven sugars are first added to dolichol pyrophosphate (PP-Dol) on the cytoplasmic face of the ER, generating Man5GlcNAc2-PP-Dol (M5GN2-PP-Dol). M5GN2-PP-Dol is then flipped across the bilayer into the lumen by an ER translocator. Genetic studies identified Rft1 as the M5GN2-PP-Dol flippase in vivo but are at odds with biochemical data suggesting Rft1 is dispensable for flipping in vitro. Thus, the question of whether Rft1 plays a direct or an indirect role during M5GN2-PP-Dol translocation has been controversial for over two decades. We describe a completely reconstituted in vitro assay for M5GN2-PP-Dol translocation and demonstrate that purified Rft1 catalyzes the translocation of M5GN2-PP-Dol across the lipid bilayer. These data, combined with in vitro results demonstrating substrate selectivity and rft1∆ phenotypes, confirm the molecular identity of Rft1 as the M5GN2-PP-Dol ER flippase. Whether Rft1 plays a role during M5GN2-PP-Dol translocation has been controversial for over two decades. In this work, a reconstituted in vitro assay demonstrates that purified Rft1 is sufficient to flip M5GN2-PP-Dol across the lipid bilayer. [ABSTRACT FROM AUTHOR]