Acid Changes Research Articles

Modulation of mercaptopurine intestinal toxicity and pharmacokinetics by gut microbiota

The interaction between the gut microbiota and mercaptopurine (6-MP), a crucial drug used in pediatric acute lymphoblastic leukemia (ALL) treatment, has not been extensively studied. Here we reveal the significant perturbation of gut microbiota after 2-week 6-MP treatment in beagles and mice followed by the functional prediction that showed impairment of SCFAs production and altered amino acid synthesis. And the targeted metabolomics in plasma also showed changes in amino acids. Additionally, targeted metabolomics analysis of feces showed changes in amino acids and SCFAs. Furthermore, ablating the intestinal microbiota by broad-spectrum antibiotics exacerbated the imbalance of amino acids, particularly leading to a significant decrease in the concentration of S-adenosylmethionine (SAM). Importantly, the depletion of gut microbiota worsened the damage of small intestine caused by 6-MP, resulting in increased intestinal permeability. Considering the relationship between toxicity and 6-MP metabolites, we conducted a pharmacokinetic study in pseudo germ-free rats to confirm that gut microbiota depletion altered the methylation metabolites of 6-MP. Specifically, the concentration of MeTINs, a secondary methylation metabolite, showed a negative correlation with SAM, the pivotal methyl donor. Additionally, we observed a strong correlation between Alistipes and SAM levels in both feces and plasma. In conclusion, our study demonstrates that 6-MP disrupts the gut microbiota, and depleting the gut microbiota exacerbates 6-MP-induced intestinal toxicity. Moreover, SAM derived from microbiota plays a crucial role in influencing plasma SAM and the methylation of 6-MP. These findings underscore the importance of comprehending the role of the gut microbiota in 6-MP metabolism and toxicity.

New approaches in pear preservation: Putrescine and modified atmosphere packaging applications to maintain fruit quality during cold storage

AbstractPear (P. communis L.), which is a climacteric fruit species, has a very short storage and shelf life, and significant losses occur due to high metabolic activity and the fruit's respiration rate after harvest. Therefore, preventing or reducing post‐harvest quality losses in pear is one of the most basic problems awaiting solution. In this study, we planned for this purpose; the fruits of the Ankara pear cultivar treated with modified atmosphere packaging (MAP), putrescine (1 mM), and MAP + putrescine were stored for 120 days at 1°C and 90 ± 5% relative humidity. The quality analyses and measurements, such as weight loss, decay rate, fruit firmness, soluble solids content (SSC), titratable acidity (TA), total phenolic compounds, antioxidant capacity, organic acids, and specific phenolic compounds, were performed on the 30th, 60th, 90th, and 120th days. Weight loss and decay ratios were lower for putrescine and putrescine + MAP‐applied fruit. With these applications, the softening of the fruit was slowed down, and the increasing SSC in the fruit and the decreasing TA rates were lower, and thus the ripening of the fruit was delayed. Changes in individual phenolic content and organic acids were lower in MAP and putrescine‐applied fruit. The study revealed that MAP and putrescine applications in pear can be used effectively to maintain fruit quality after harvest.

Combining multivariate statistical analysis to characterize changes in amino acids and volatiles during growth of Lou onion pseudostems.

The main edible part of the Lou onion is the pseudostem, which is highly valued for its distinctive flavour. However, harvesting decisions for the pseudostem are often based on size and market price, with little consideration given to flavour. By clarifying the growth of flavour in pseudostems, farmers and consumers may benefit from evidence-based insights that help optimize harvesting time and maximize flavour quality. This study employed amino acid analysis and gas chromatography-ion migration spectroscopy (GC-IMS) to elucidate the compounds of the pseudostem across different growth phases, and 17 amino acids and 61 volatile substances. Subsequently, analysis revealed that 18 compounds, including arginine (Arg), aspartic acid (Asp), glutamic acid (Glu), valine (Val), (E)-2-nonenal, decanal, 2,4-nonadienal, 2-octenal, (Z)-4-decenal, 2,4-decadienal benzeneacetaldehyde, linalool, eugenol, (Z)-6-nonen-1-ol, methyl anthranilate, 2-acetylpyridine, 3-sec-butyl-2-methoxypyrazine, and 2,6-dichlorophenol, were the key compounds in determining the flavour characteristics of the pseudostems, as assessed by taste activity value and relative odour activity value calculations. In addition, correlation analysis, focusing on five amino acids and 38 volatile compounds with variable importance for predictive components scores of >1, identified anisaldehyde, eugenol, (Z)-6-nonen-1-ol, 2,4-decadienal, 3-sec-butyl-2-methoxypyrazine, Arg, Asp, and Val as the key differentiators and contributors to the pseudostems flavour profile. During the rapid growth of Lou onions just before the emergence of flower stems, the pseudostem exhibited the most prominent flavour, making this stage most suitable for harvesting compared to the regreening growth stage and the rapid growth period of the aerial bulbs. © 2024 Society of Chemical Industry.

Changes in Plasma Free Amino Acid Profile in Endurance Athletes over a 9-Month Training Cycle.

We aimed to evaluate long-term changes in proteinogenic and non-proteinogenic plasma free amino acids (PFAA). Eleven male endurance triathletes participated in a 9-month study. Blood was collected at rest, immediately after exhaustive exercise, and during 30-min recovery, in four consecutive training phases: transition, general, specific, and competition. Twenty proteinogenic and 22 non-proteinogenic PFAAs were assayed using the LC-ESI-MS/MS technique. The structured training modified the patterns of exercise-induced PFAA response, with the competition phase being the most distinct from the others. Branched-chain amino acids (p = 0.002; η2 = 0.216), phenylalanine (p = 0.015; η2 = 0.153), methionine (p = 0.002; η2 = 0.206), and lysine (p = 0.006; η2 = 0.196) declined more rapidly between rest and exhaustion in the competition phase. Glutamine (p = 0.008; η2 = 0.255), glutamate (p = 0.006; η2 = 0.265), tyrosine (p = 0.001; η2 = 0.195), cystine (p = 0.042; η2 = 0.183), and serine (p < 0.001; η2 = 0.346) levels were reduced in the competition phase. Arginine (p = 0.046; η2 = 0.138) and aspartate (p = 0.011; η2 = 0.171) levels were highest during exercise in the transition phase. During the competition phase, α-aminoadipic acid (p = 0.023; η2 = 0.145), β-aminoisobutyric acid (p = 0.007; η2 = 0.167), β-alanine (p < 0.001; η2 = 0.473), and sarcosine (p = 0.017; η2 = 0.150) levels increased, whereas phosphoethanolamine (p = 0.037; η2 = 0.189) and taurine (p = 0.008; η2 = 0.251) concentrations decreased. Overtraining indicators were not elevated. The altered PFAA profile suggests adaptations within energy metabolic pathways such as the tricarboxylic acid cycle, oxidative phosphorylation, ammonia neutralization, the purine nucleotide cycle, and buffering of intracellular H+ ions. The changes seem to reflect normal adaptations.

Assessing the influence of nano graphene oxide application in mitigating the adverse effects of runoff pH on the moisture susceptibility of asphalt concretes

The specifications of runoff entered the pavement has an essential role in the severity of moisture damage of asphalt concretes. Changing the pH of runoff has a negative impact on moisture damage and intensifies this damage in asphalt concretes. Accordingly, in this study, the impact of nanographene oxide (NGO) as a contaminant adsorbent and bitumen modified to decrease the destructive impacts of acidity changes in runoff has been investigated. For this purpose, limestone and siliceous aggregates, PG 64–16 bitumen, and NGO in two different contents (0.3 % and 0.5 % of the weight of bitumen) were utilized. To asses the strength of asphalt concretes with different runoff pH conditions, hydrochloric acid and sodium oxide were used to simulate acidic and basic runoffs, respectively. The indirect tensile strength (ITS) of asphalt concretes in different environmental states was tested to compare the severity of moisture sensitivity. Moreover, the surface free energy (SFE) elements of bitumen and aggregates were calculated utilizing the Wilhelmy Plate (WP) and the Universal Sorption Device (USD), respectively. The parameters of SFE theory were used to assess the mechanisms of water damage potential. The SFE results showed that NGO decreased the debonding energy (DE) of the mixtures. Changes in DE in neutral environments were minor, but with the polarization of environmental conditions, especially in basic states, these changes intensified. The application of NGO improved the total bitumen SFE, which reduced the possibility of moisture sensitivity as a cohesive failure. The results of mechanical methods indicated that the use of NGO in specimens with limestone and siliceous aggregate increased the indirect tensile strength ratio (TSR) in moisture conditions with different degrees of acidity. The mechanical experiment results also confirmed the SFE method's results that the effect of using NGO on increasing the TSR index at pH equal to 7 was minor, and it was intensified in acidic and basic waters.

Butyrate- and Beta-Hydroxybutyrate-Mediated Effects of Interventions with Pro- and Prebiotics, Fasting, and Caloric Restrictions on Depression: A Systematic Review and Meta-Analysis.

To examine the butyrate- and beta-hydroxybutyrate (BHB)-modulated effects of pre- and probiotic interventions, fasting, and caloric restriction interventions, a systematic literature review was carried out with a subsequent meta-analysis. Three pre-and probiotic intervention randomized control trials (RCTs) were included in the meta-analysis. A significant increase in butyrate (standardized mean difference (SMD) [confidence interval (CI)] 0.34; [0.02-0.67]) and an improvement in depression scores (SMD [CI] 0.15, [-0.35-0.70]) through pre- and probiotic interventions were shown in the meta-analysis. The intervention duration of the included studies ranged from three days to four weeks, with the examined population being healthy adults. Butyrate was measured in either plasma or feces, and the depression score was obtained under the Swedish core affect scale, the hospital anxiety and depression scale (HADS), or the depression, anxiety, and stress scale-21 items (DASS-21). In addition to butyrate, the total SCFA concentration also seems to be positively associated with pre- and probiotic administration (SMD [CI] 0.55 [0.15-0.95]). Despite the significant short-chain fatty acid (SCFA) and butyrate concentration changes, no significant correlation between butyrate and depression or between SCFAs and depression could be shown through linear regression models. Nevertheless, the regression coefficient b1 = 1.57 (p = 0.17) for butyrate suggests a strong, positive connection between butyrate and depression. Additionally, three studies were qualitatively analyzed, examining fasting as an intervention and revealing a connection between fasting, BHB, and depression. The association between fasting, BHB, and depression or mood elevation appeared to be related to BHB concentrations, which may be due to the similar biochemical properties of BHB and butyrate. Furthermore, caloric restrictions as alternatives to fasting were proposed as potential long-term interventions.

The First In Vivo Study Shows That Gyrophoric Acid Changes Behavior of Healthy Laboratory Rats.

Gyrophoric acid (GA), a lichen secondary metabolite, has attracted more attention during the last years because of its potential biological effects. Until now, its effect in vivo has not yet been demonstrated. The aim of our study was to evaluate the basic physicochemical and pharmacokinetic properties of GA, which are directly associated with its biological activities. The stability of the GA in various pH was assessed by conducting repeated UV-VIS spectral measurements. Microsomal stability in rat liver microsomes was performed using Ultra-Performance LC/MS. Binding to human serum albumin (HSA) was assessed using synchronous fluorescence spectra, and molecular docking analysis was used to reveal the binding site of GA to HSA. In the in vivo experiment, 24 Sprague-Dawley rats (Velaz, Únetice, Czech Republic) were used. The animals were divided as follows. The first group (n = 6) included healthy males as control intact rats (♂INT), and the second group (n = 6) included healthy females as controls (♀INT). Groups three and four (♂GA/n = 6 and ♀GA/n = 6) consisted of animals with daily administered GA (10 mg/kg body weight) in an ethanol-water solution per os for a one-month period. We found that GA remained stable under various pH and temperature conditions. It bonded to human serum albumin with the binding constant 1.788 × 106 dm3mol-1 to reach the target tissue via this mechanism. In vivo, GA did not influence body mass gain, food, or fluid intake during the experiment. No liver toxicity was observed. However, GA increased the rearing frequency in behavioral tests (p < 0.01) and center crossings in the elevated plus-maze (p < 0.01 and p < 0.001, respectively). In addition, the time spent in the open arm was prolonged (p < 0.01 and p < 0.001, respectively). Notably, GA was able to pass through the blood-brain barrier, indicating its ability to permeate into the brain and to stimulate neurogenesis in the hilus and subgranular zone of the hippocampus. These observations highlight the potential role of GA in influencing brain function and neurogenesis.

Signal-Sustained Imaging of Mitophagy with an Enzyme-Activatable Metabolic Lipid Labeling Probe

ABSTRACT Imaging of mitophagy is of significance as aberrant mitophagy is engaged in multiple diseases. Mitophagy has been imaged with synthetic or biotic pH sensors by reporting pH acidification en route delivery into lysosomes. To circumvent uncertainty of acidity-dependent signals, we herein report an enzyme-activatable probe covalently attached on mitochondrial inner membrane (ECAM) for signal-persist mitophagy imaging. ECAM is operated via ΔΨm-driven accumulation of Mito-proGreen in mitochondria and covalent linking of the trapped probe with azidophospholipids metabolically incorporated into the mitochondrial inner membrane. Upon mitophagy, ECAM is delivered into lysosomes and hydrolyzed by LNPEP/leucyl aminopeptidase, yielding turn-on green fluorescence that is immune to lysosomal acidity changes and stably retained in fixed cells. With ECAM, phorbol-12-myristate-13-acetate (PMA) was identified as a highly potent inducer of mitophagy. Overcoming signal susceptibility of pH probes and liability of ΔΨm probes to dissipation from stressed mitochondria, ECAM offers an attractive tool to study mitophagy and mitophagy-inducing therapeutic agents. Abbreviations: Baf-A1, bafilomycin A1; CCCP, carbonyl cyanide m-chlorophenylhydrazone; DBCO, dibenzocyclooctyne; ECAM, enzyme-activated probe covalently attached on mitochondrial inner membrane; GFP, green fluorescent protein; LAMP2, lysosomal associated membrane protein 2; LNPEP/LAP, leucyl and cystinyl aminopeptidase; PMA, phorbol-12-myristate-13-acetate; ΔΨm, mitochondrial transmembrane potential; RFP, red fluorescent protein; TPP, triphenylphosphonium.

Identification and Molecular Analysis of Hemocyanin as a Body Resistance Gene in Litopenaeus vannamei

Production of white shrimp (Litopenaeus vannamei) tends to decrease every year caused by diseases, one of which is White Feces Disease (WFD). Genetically, monitoring has been carried out on the parent characters of SPF (Specific Pathogen Free) shrimp imported from Hawaii and Florida with low genetic variation. If there is a threat such as disease caused by bacteria and the gene that codes for disease control or the body's resistance gene is missing, it allows the individual to become susceptible to disease. Gene that codes for body resistance in white shrimp is hemocyanin. Hemocyanin gene studies are needed as a first step in controlling disease in white shrimp. Method used is a descriptive method. Procedures carried out are sample collection, detection of WFD, molecular analysis of hemocyanin and data analysis. Results showed that healthy shrimp had hemocyanin while shrimp with WFD had very little or no amount of hemocyanin. Hemocyanin gene profile in healthy shrimp was also analyzed and there were 4 nucleotide change made changes to 4 amino acid. Changes in amino acids affect the stability of hemocyanin and cause polymorphism of hemocyanin. Phylogenetic analysis showed dendogram split into two clades between L. vannamei and P. monodon.

Establishment and evaluation of a qPCR method for the detection of pfmdr1 mutations in Plasmodium falciparum, the causal agent of fatal malaria

Drug resistance surveillance is a major integral part of malaria control programs. Molecular methods play a pivotal role in drug resistance detection and related molecular research. This study aimed to develop a rapid and accurate detection method for drug resistance of Plasmodium falciparum (P. falciparum). A quantitative real-time PCR (qPCR) assay has been developed that identifies the mutation at locus A256T in the P.falciparum multi-drug resistance(pfmdr1) gene producing amino acid change at position 86. The results of 198 samples detected by qPCR were consistent with nested PCR and sequencing, giving an accuracy of 94.3%. The sensitivity, specificity, positive and negative predictive value of qPCR were 85.7%, 97.6%, 90.0% and 96.4%, respectively. The results of qPCR are basically consistent with the nested PCR, which is expected to replace the nested PCR as a new molecular biological method for drug resistance detection, providing reliable technical support for global malaria prevention and control.

Characterization of the plasma metabolome and lipidome in response to sleeve gastrectomy and gastric bypass surgeries reveals molecular patterns of surgical weight loss

ObjectivesBariatric surgery improves metabolic health, but the underlying mechanisms are not fully understood. We analyzed the effects of two types of bariatric surgery, sleeve gastrectomy (SG) and Roux-en-Y gastric bypass (RYGB), on the plasma metabolome and lipidome. MethodsWe characterized the plasma metabolome (1268 metabolites) and lipidome (953 lipids) pre-operatively and at 3 and 12 months post-operatively in 104 obese adults who were previously recruited to a prospective cohort of bariatric surgery. The metabolomic and lipidomic responses to bariatric surgery over time were analyzed using multivariable linear mixed-effects models. ResultsThere were significant changes in multiple metabolites and lipids, including rapid early changes in amino acid and peptide metabolites, including decreases in branched-chain amino acids (BCAAs), aromatic AAs, alanine and aspartate, and increases in glycine, serine, arginine and citrulline. There were also significant decreases in many triglyceride species, with increases in phosphatidylcholines and phosphatidylethanolamines. There were significant changes in metabolites related to energy metabolism that were apparent only after 12 months. We observed differences by bariatric surgery type in the changes in a small number of primary and secondary bile acids, including glycohyocholate and glyco-beta-muricholate. ConclusionsOur findings highlight the comprehensive changes in metabolites and lipids that occur over the 12 months following bariatric surgery. While both SG and RYGB caused profound changes in the metabolome and lipidome, RYGB was characterized by greater increases in bile acids following surgery.

RBD design increases the functional antibody titers elicited by SARS-CoV-2 spike vaccination

Most COVID-19 vaccines contain the SARS-CoV-2 spike protein as an antigen, but they lose efficacy as neutralizing antibody titers wane and escape variants emerge. Modifying the spike antigen to increase neutralizing antibody titers would help counteract this decrease in titer. We previously used a structure-based computational design method to identify nine amino acid changes in the receptor-binding domain (RBD) of spike that stabilize the RBD and increase the neutralizing antibody titers elicited by vaccination. Here, we introduce those enhancing amino acid changes into a full-length spike (FL-S-2P) ectodomain representative of most approved vaccine antigens. These amino acid changes can be incorporated into the FL-S-2P protein without negatively effecting expression or stability. Furthermore, the amino acid changes improved functional antibody titers in both mice and monkeys following vaccination. These amino acid changes could increase the duration of protection conferred by most COVID-19 vaccines.

Analysis of CYP1B1 Polymorphisms in Lung Cancer Patients Using Novel, Quick and Easy Methods Based on CAPS and ACRS-PCR Techniques.

Within the sequence of the CYP1B1 gene, more than 50 polymorphisms, resulting from single-nucleotide polymorphisms (SNPs), have been described. Some of them play an important role as specific genetic markers in the process of carcinogenesis and for therapeutic purposes. In this publication, we present methods we have developed that enable the specific and unambiguous identification of four polymorphisms that result in amino acid changes: c. 142C > G, c. 355G > T, c. 1294C > G, and c. 1358A > G. Our studies are based on cleaved amplified polymorphic sequences (CAPSs) and artificially created restriction site (ACRS) PCR techniques; therefore, they require only basic laboratory equipment and low financial outlays. Utilizing the described methods allows for the reduction of research time and cost, and the minimization of errors. Their effectiveness and efficiency depend on the careful design of appropriate primers and the precise selection of suitable restriction enzymes. As a result, further confirmation by sequencing is not necessary. Using the developed method, we examined 63 patients diagnosed with lung cancer and observed a 1.5 to 2.1 times higher frequency of the analyzed single-nucleotide polymorphisms compared to the frequency in the European population.

Intensification of the fermentation process of a plant-based drink with complex starter «Bifido Plus»

Relevance. Developing the concept of optimal healthy nutrition increases the demand for the development and production of fortified and functional food products. We propose to use plant-based drinks as a food system for obtaining a probiotic product, which is a source of a number of nutrients. Because the environment in plants isn’t usually good for growing lactic acid bacteria, it’s important to look into different ways to start lactic acid fermentation in these kinds of places.The purpose of these studies is to evaluate ways to intensify the fermentation process of a vegetable drink with a complex starter culture «Bifido plus».Мethods. Research methods include assessment of the increase in microbial biomass, accumulation of lactic acid and exopolysaccharides, and changes in acidity and viscosity of the probiotic drink.Results. The possibility of adaptation of the starter «Bifido Plus» in a plant environment was established, and ultrasonic exposure and the addition of inulin promoted the activation of the fermentation process. In particular, the addition of inulin shortens the lag phase by 0.85 ± 0.05 hours, and exposure to ultrasound treatment increases it by 2.1 ± 0.2 hours. At the same time, the subsequent increase in the volume of biomass accumulation in the sample of the probiotic drink subjected to ultrasound exposure was 60–313% and 28–44% under the influence of inulin. The introduction of inulin and exposure to ultrasound also activate a change in titratable acidity, on average, by 3–15%, the accumulation of lactic acid by 38.7–68%, and the accumulation of exopolysaccharides by 27.3–58.8% relative to the control sample. So, the research results show that ultrasound and inulin can be used to start the fermentation process in plant media. The research was supported by a grant from the Russian Science Foundation (RSF) within the framework of project 23-26-10063.

Bioconversion of mangosteen pericarp extract juice with bacterial cellulose production and probiotic integration: A promising approach for functional food supplement development

This study optimized bioconversion process of mangosteen pericarp extract juice (MPEj) with a single-culture of Komagataeibacter xylinus TISTR 1061, a bacterial cellulose-producing strain and a co-culture with the probiotic Lactiplantibacillus plantarum. K. xylinus TISTR 1061 was identified as the optimal bacterial cellulose-producing strain for fermenting MPEj. Optimal conditions were determined to be 20% (v/v) MPEj supplemented with 7% (w/v) sucrose and 0.2% (w/v) ammonium dihydrogen phosphate as carbon and nitrogen sources, respectively. The growth profile of single K. xylinus was not significantly different from those of the co-culture. They showed good adaptability and cohabited during fermentation without antagonistic effects. Dynamic changes in total titratable acidity (TTA), total soluble solids (TSS), and pH during fermentation were relatively similar for both single- and co-cultures. Post-fermented MPEj with co-culture exhibited significantly higher bioactive activities, particularly antioxidant properties. Significant variations in metabolite concentrations, including 2-aminobutyrate, valine, xylitol, propionate, isoleucine, glucose, and glycerol, were observed. These compounds exhibited significantly higher average concentrations after fermentation, highlighting their potential as biomarkers distinguishing pre- and post-fermentation MPEj. Bacterial cellulose (BC) from single culture exhibited greater thickness and weight, with a more complex and branched structure, while that from co-culture was infused with probiotic cells, showing less complexity allowing the BC from co-culture demonstrated low hardness and chewiness values. The properties of both bioconverted MPEj and BC obtained as observed offer potential for innovative functional drinks and/or dietary supplements with enhanced health benefits.

Cloning, Expression, Enzymatic Characterization and Mechanistic Studies of M13 Mutant Acetohydroxyacid Synthase That Rescues Valine Feedback Inhibition

Acetohydroxyacid synthase (AHAS) is a key enzyme in the first step of the branched-chain amino acid synthesis pathway, and the production of acetohydroxybutyrate from one molecule of 2-ketobutyric acid and one molecule of pyruvate. AHAS is inhibited by feedback from L-valine, L-leucine, and L-isoleucine, and the expression of ilvBN, the gene encoding AHAS, is regulated by all three branched-chain amino acids. A change in amino acids 20–22 on the regulatory subunit (M13 mutation) removes the feedback inhibition by valine. We cloned the gene encoding AHAS (ilvBN) into a vector and then transfected it into Escherichia coli BL21 for expression with targeted changes in amino acids 20–22 on the regulatory subunit, and then determined the activity of the mutated AHAS and its inhibitory effects on valine, isoleucine, and leucine. The enzyme containing the M13 mutation was feedback resistant to all three amino acids. Previous studies have suggested that the binding sites for the three branched-chain amino acids may be at the same variable center. We investigated the enzymatic properties of wild-type and mutant AHAS, modeled their crystal structures, and resolved the mechanism of feedback inhibition induced by mutant M13, which will be useful for continuing the modification of AHAS and the design of broad-spectrum herbicides.

Biomarkers for ideal protein: rabbit diet metabolomics varying key amino acids

With the main aim of identifying biomarkers that contribute to defining the concept of ideal protein in growing rabbits under the most diverse conditions possible this work describes two different experiments. Experiment 1: 24 growing rabbits are included at 56 days of age. The rabbits are fed ad libitum one of the two experimental diets only differing in lysine levels. Experiment 2: 53 growing rabbits are included at 46 days of age, under a fasting and eating one of the five experimental diets, with identical chemical composition except for the three typically limiting amino acids (being fed commercial diets ad libitum in both experiments). Blood samples are taken for targeted and untargeted metabolomics analysis. Here we show that the metabolic phenotype undergoes alterations when animals experience a rapid dietary shift in the amino acid levels. While some of the differential metabolites can be attributed directly to changes in specific amino acids, creatinine, urea, hydroxypropionic acid and hydroxyoctadecadienoic acid are suggested as a biomarker of amino acid imbalances in growing rabbits’ diets, since its changes are not attributable to a single amino acid. The fluctuations in their levels suggest intricate amino acid interactions. Consequently, we propose these metabolites as promising biomarkers for further research into the concept of the ideal protein using rabbit as a model.

Simultaneous removal of ammonium, phosphate, and phenol via self-assembled biochar composites CBCZrOFe3O4 and its utilization as soil acidity amelioration

ABSTRACT High concentrations of ammonium, phosphate, and phenol are recognized as water pollutants that contribute to the degradation of soil acidity. In contrast, small quantities of these nutrients are essential for soil nutrient cycling and plant growth. Here, we reported composite materials comprising biochar, chitosan, ZrO, and Fe3O4, which were employed to mitigate ammonium, phosphate, and phenol contamination in water and to lessen soil acidity. Batch adsorption experiments were conducted to assess the efficacy of the adsorbents. Initially, comparative studies on the simultaneous removal of NH4, PO4, and phenol using CB (biochar), CBC (biochar + chitosan), CBCZrO (biochar + chitosan + ZrO), and CBCZrOFe3O4 (biochar + chitosan + ZrO + Fe3O4) were conducted. The results discovered that CBCZrOFe3O4 exhibited the highest removal percentage among the adsorbents (P < 0.05). Adsorption data for CBCZrOFe3O4 were well fitted to the second-order kinetic and Freundlich isotherm models, with maximum adsorption capacities of 112.65 mg/g for NH4, 94.68 mg/g for PO4 and 112.63 mg/g for phenol. Subsequently, the effect of CBCZrOFe3O4-loaded NH4, PO4, and phenol (CBCZrOFe3O4-APP) on soil acidity was studied over a 60-day incubation period. The findings showed no significant changes (P < 0.05) in soil exchangeable acidity, H+, Mg, K, and Na. However, there was a substantial increase in the soil pH, EC, available P, CEC, N-NH4, and N-NO3. A significant reduction was also observed in the available soil exchangeable Al and Fe (P < 0.05). This technique demonstrated multi-functionality in remediating water pollutants and enhancing soil acidity.

Biological and molecular characterization of pydiflumetofen and phenamacril dual-resistant Fusarium graminearum strains.

Fusarium head blight (FHB) caused by Fusarium graminearum species complex (FGSG) remains a major challenge to cereal crops and resistance to key fungicides by the pathogen threatens control efficacy. Pydiflumetofen, a succinate dehydrogenase inhibitor, and phenamacril, a cyanoacrylate fungicide targeting myosin I, have been applied to combat this disease. Nonetheless, emergence of pydiflumetofen resistance in a subset of field isolates alongside laboratory-induced facile generation of phenamacril-resistant isolates signals a critical danger of resistance proliferation. Our study investigates the development of dual resistance to these fungicides in F. graminearum. Utilizing pydiflumetofen-resistant (PyR) and -sensitive (PyS) isolates, we obtained dual-resistant (PyRPhR) and phenamacril-resistant (PySPhR) mutants on potato sucrose agar containing phenamacril. Mutation rates for phenamacril resistance were comparable between pydiflumetofen-resistant and -sensitive isolates, implying independent pathways for resistance development. The mutants compromised in fungal growth, competitive viability and deoxynivalenol production, suggesting fitness penalties for the dual-resistant mutants. However, no cross-resistance was found with tebuconazole or fludioxonil. In addition, we characterized four critical amino acid changes (S217L, C423R, K537T, E420G) in the Myo1 that were verified to confer phenamacril resistance in F. graminearum. This research indicates the possibility of resistance development for both pydiflumetofen and phenamacril in F. graminearum and emphasizes the need for fungicide resistance management for FHB. © 2024 Society of Chemical Industry.

Rapid profiling of fish cell nitrogen metabolism with single-cell Raman spectroscopy: Unveiling enzyme's role in ammonia detoxification

Ammonia is a prevalent aquatic pollutant that disrupts cellular functions and energy metabolism in fish, posing significant environmental and health threats. This research investigates the critical role of arginase 2 (ARG2) in mitigating ammonia toxicity in fish cells and its implications in adapting to nitrogen metabolism under high ammonia exposure. Through a CRISPR-Cas9 engineered ARG2 knockdown (KD) in the Epithelioma Papulosum Cyprini (EPC) cell line, we first investigated the biochemical responses of ARG2 KD and wild-type (WT) EPC cells to ammonia stress (NH4Cl treatment), showing diminished urea production and decreased cell viability in ARG2 KD cells. Subsequently, single-cell Raman spectroscopy analysis revealed that ARG2 KD cells exhibited profound metabolic shifts, including changes in protein, nucleic acids, lipid and sugar levels, showing the adjusting role of ARG2 in the balance of carbohydrate and nitrogen metabolism. Furthermore, the upregulated responses of various amino acids, such as glutamine, arginine, alanine, glutamic acid, glycine, histidine, phenylalanine and valine, in WT cells after NH4Cl treatment diminished in ARG2 KD cells except for the decrease in aspartic acid, indicating a switching effect of ARG2 in nitrogen metabolism under ammonia stress. This study highlights ARG2's essential role in ammonia detoxification and emphasizes ARG2's protective function and its importance in metabolism, shedding light on the adaptive mechanisms fish cells deploy against high ammonia environments. These insights contribute to deep understanding of aquatic organisms' molecular responses to environmental ammonia pollution, offering potential strategies for their protection.