Major Amino Acids Research Articles

Isolation and characterization of nettle (Urtica dioica L.) seed proteins: Conversion of underutilized by-products of the edible oil industry into food emulsifiers

This study aimed to upcycle a byproduct of the edible oil industry, cold-pressed nettle seed meal (CPNSM), into a plant-based emulsifier, thereby increasing the sustainability of the food system. The protein content of the nettle seed protein (NSP) powder was 48.3% with glutamic acid (16.6%), asparagine (10.7%), and arginine (9.7%) being the major amino acids. NSPs had a denaturation temperature of 66.6 °C and an isoelectric point of pH 4.3. They could be used as emulsifiers to form highly viscous coarse corn oil-in-water emulsions (10% oil, 4% NSP). Nevertheless, 10-fold diluted emulsions exhibited rapid creaming under different pH (2–9), salt (0–500 mM NaCl) and temperature (>40 °C) conditions, but they were relatively stable to aggregation. Our findings suggest that NSPs could be used as emulsifiers in highly viscous or gelled foods, like dressings, sauces, egg, cheese, or meat analogs.

Changes in the Composition of Olive Pomace after Fermentation: A Preliminary Study

Because olive pomace (the main by-product of olive oil production) is phytotoxic, new applications must be investigated to minimize its negative environmental impact. In this work, olive pomace was fermented for 4 and 32 days at room temperature, having in view its valorization as a novel food, thereby creating opportunities for the food industry and addressing a challenge of the olive oil sector. The chemical and microbiological modifications that occurred along the fermentation were followed. The results showed no significant differences (p > 0.05) in total protein between the control and the fermented samples; however, the latter exhibited higher levels of essential amino acids. The major nonessential and essential amino acids were glutamic acid and leucine in all samples. There was a significant increase in the total fat of the 32-day sample and the main fatty acid was oleic acid in all samples. There were considerable reductions in total vitamin E, phenolics, and antioxidant activity values post-fermentation. Hydroxytyrosol replaced oleacein as the main phenolic in the 32-day sample. A sharp increase in total microorganisms occurred (2.20 × 102 to 3.00 × 104–2.01 × 107 colony forming units/mL) but no pathogenic microorganisms were detected. Overall, olive pomace fermentation creates novel products for the food industry with a balanced nutritional composition.

Formation of Cysteine Adducts with Chlorogenic Acid in Coffee Beans.

The post-harvest processing of coffee beans leads to a wide range of reactions involving proteins. The formation of crosslinks between proteins and phenolic compounds present in high concentrations of coffee beans represents one of the most challenging and still not fully characterized reactions. The aim of this work was to assess the presence of products from such reactions in coffee samples, focusing on the adducts between cysteine and chlorogenic acids (CQAs). For this purpose, 19 green and 15 roasted coffee samples of the Coffea arabica, Coffea canephora, and Coffea liberica varieties were selected for this study and basically characterized. Then, targeted liquid chromatography mass spectrometry (LC-MS/MS) methods were developed to assess the formation of adducts between CQA and cysteine, glutathione, and N-acetylcysteine as the amino acid and peptide models, and quantified such adducts in coffee samples. The results of the characterization showed a heterogeneous distribution of the protein content (8.7-14.6%), caffeine (0.57-2.62 g/100 g), and antioxidant capacity (2-4.5 g ascorbic acid/100 g) in Arabica, Canephora, and Liberica samples. Glutamic acid, arginine, and proline were found to be the major amino acids, while 5-CQA (38-76%), 3-CQA (4-13%), and 4-CQA (4-13%) were the most abundant CQA derivatives of all coffee varieties. The model experiments for adduct formation demonstrated that cysteine binds to CQA via thiol groups and 5-CQA initially isomerizes to 3- and 4-CQA, depending on the conditions, allowing cysteine to bind to two different sites on 3-, 4- or 5-CQA molecules, thus, forming six different Cys-CQA adducts with m/z 476. The reaction was more favored at pH 9, and the adducts proved to be stable up to 90 °C for 10 min and up to 28 days at room temperature. The relative quantification of adducts showed peak area values ranging from 1100 to 3000 in green coffee bean samples, while no adducts were detected in roasted coffee beans. Overall, this work was the first attempt to demonstrate the presence of Cys-CQA adducts in coffee beans and paves the way for further investigations of such adduct formation at the protein level.

Comparative Metabolome Profiling for Revealing the Effects of Different Cooking Methods on Glutinous Rice Longjing57 (Oryza sativa L. var. Glutinosa).

Glutinous rice (GR), an important food crop in Asia, provides prolonged energy for the human body due to its high amylopectin content. The non-volatile metabolites generated by different cooking methods that affect the nutritional value and color of GR are still poorly understood. Herein, a widely targeted metabolomics approach was used to understand the effects of different cooking methods (steaming, baking, and frying) on the metabolite profiles of GR. Compared with other treatments, steamed GR had a brighter color and significantly lower contents of total sugar, starch, amylopectin, and amylose, at 40.74%, 14.13%, 9.78%, and 15.18%, respectively. Additionally, 70, 108, and 115 metabolites were significantly altered in the steaming, baking, and frying groups respectively, and amino acid and carbohydrate metabolism were identified as the representative metabolic pathways based on KEGG annotations. Further evaluation of 14 amino acids and 12 carbohydrates in steamed GR, especially 4-aminobutyric acid, suggested its high nutraceutical value. Additionally, multivariate analysis indicated that total sugar content, amylose content, beta-alanine methyl ester hydrochloride, and 4-aminobutyric acid played a critical role in color formation in raw and cooked GR. Finally, the levels of major amino acids and carbohydrates were quantified by conventional methods to verify the reliability of the metabolome. Consequently, this in-depth understanding of metabolite profiling in normal cooking methods has provided a foundation for the processing of GR products.

Genomic analysis and phylogenetic characterization of Himalayan snow trout, Schizothorax esocinus based on mitochondrial protein-coding genes.

Mitochondrial DNA (mtDNA) has become a significant tool for exploring genetic diversity and delineating evolutionary links across diverse taxa. Within the group of cold-water fish species that are native to the Indian Himalayan region, Schizothorax esocinus holds particular importance due to its ecological significance and is potentially vulnerable to environmental changes. This research aims to clarify the phylogenetic relationships within the Schizothorax genus by utilizing mitochondrial protein-coding genes. Standard protocols were followed for the isolation of DNA from S. esocinus. For the amplification of mtDNA, overlapping primers were used, and then subsequent sequencing was performed. The genetic features were investigated by the application of bioinformatic approaches. These approaches covered the evaluation of nucleotide composition, codon usage, selective pressure using nonsynonymous substitution /synonymous substitution (Ka/Ks) ratios, and phylogenetic analysis. The study specifically examined the 13 protein-coding genes of Schizothorax species which belongs to the Schizothoracinae subfamily. Nucleotide composition analysis showed a bias towards A + T content, consistent with other cyprinid fish species, suggesting evolutionary conservation. Relative Synonymous Codon Usage highlighted leucine as the most frequent (5.18%) and cysteine as the least frequent (0.78%) codon. The positive AT-skew and the predominantly negative GC-skew indicated the abundance of A and C. Comparative analysis revealed significant conservation of amino acids in multiple genes. The majority of amino acids were hydrophobic rather than polar. The purifying selection was revealed by the genetic distance and Ka/Ks ratios. Phylogenetic study revealed a significant genetic divergence between S. esocinus and other Schizothorax species with interspecific K2P distances ranging from 0.00 to 8.87%, with an average of 5.76%. The present study provides significant contributions to the understanding of mitochondrial genome diversity and genetic evolution mechanisms in Schizothoracinae, hence offering vital insights for the development of conservation initiatives aimed at protecting freshwater fish species.

Analysis of plasma metabolomes from 11 309 subjects in five population-based cohorts

Plasma metabolomics holds potential for precision medicine, but limited information is available to compare the performance of such methods across multiple cohorts. We compared plasma metabolite profiles after an overnight fast in 11,309 participants of five population-based Swedish cohorts (50–80 years, 52% women). Metabolite profiles were uniformly generated at a core laboratory (Metabolon Inc.) with untargeted liquid chromatography mass spectrometry and a comprehensive reference library. Analysis of a second sample obtained one year later was conducted in a subset. Of 1629 detected metabolites, 1074 (66%) were detected in all cohorts while only 10% were unique to one cohort, most of which were xenobiotics or uncharacterized. The major classes were lipids (28%), xenobiotics (22%), amino acids (14%), and uncharacterized (19%). The most abundant plasma metabolome components were the major dietary fatty acids and amino acids, glucose, lactate and creatinine. Most metabolites displayed a log-normal distribution. Temporal variability was generally similar to clinical chemistry analytes but more pronounced for xenobiotics. Extensive metabolite-metabolite correlations were observed but mainly restricted to within each class. Metabolites were broadly associated with clinical factors, particularly body mass index, sex and renal function. Collectively, our findings inform the conduct and interpretation of metabolite association and precision medicine studies.

Predominant amino acid substitutions in NS5B gene of hepatitis C virus in blood donors and treatment-naïve hepatitis and HIV patients in Nigeria

Background: Hepatitis C virus (HCV) genome undergoes high rate of mutation, which results in generation of genetically diverse HCV isolates. There is paucity of data on mutations in the nonstructural 5b (NS5b) gene of circulating HCV and their implications in the Nigerian population. Here, we identified clinically-important mutations in HCV isolates, which may influence response to therapy and disease prognosis.
 Methodology: HCV RNA was extracted from a total of 301 blood samples collected from 99 symptomatic treatment-naïve hepatitis patients, 125 HIV-infected individuals and 77 asymptomatic blood donors in Ibadan, Nigeria. The RNA was reverse–transcribed to complimentary DNA and HCV NS5B gene amplified by nested PCR. The amplified products of 42 HCV were sequenced and sequences were aligned with those from GenBank and HCV databases in MEGA 7.0. Nucleotide sequences were translated to amino acids while substitutions in the amino acids were analyzed with reference to H77 prototype strain of HCV.
 Results: A total of 10 amino acid polymorphisms were observed from the 42 sequenced NS5B gene, with the major clinically-important amino acid mutations being S15G in 28 (66.7%) participants, T7N (24, 57.1%), G61R (23, 54.8%), S54L (22, 52.4%), G89E (14, 33.3%), T79M (12, 28.6%), and T711 (11, 26.2%). Others were Q67R (7, 16.7%), Q47H (7, 16.7%) and S84F (2, 4.8%). S15G/A/V mutations were more predominant in patients with HIV (76.9%, 10/13) followed by patients with clinical hepatitis (75.0%, 12/16) and blood donors (46.1%, 6/13). Q67R and T71I mutations were not predominant in patients with clinical hepatitis as they were detected in only 31.3% (5/16) and 43.8% (7/16) participants respectively, compared to S15G (75.0%, 12/16), S54L (68.8%, 11/16), G61R/E (68.8%, 11/16) and T7N/S (56.3%, 9/16). There was no statistically significant difference in the distribution of each of the 10 amino acid polymorphisms detected within patients with symptomatic clinical hepatitis (x2=9.311, p=0.409), HIV-infected patients (x2=13.431, p=0.1440) and asymp- tomatic blood donors (x2=3.775, p=0.9256). Similarly, there was no significant difference in the distribution between the 3 categories of the study participants except for T79M mutation, which was significantly higher in HIV-infected patients (61.5%, 8/13) compared to patients with clinical hepatitis (18.8%, 3/16) and asymp- tomatic blood donors (7.7%, 1/13) (x2=10.456, p=0.0054).
 Conclusion: Mutations in the NS5B gene could be associated with worse prognosis of the disease or antiviral failure due to viral resistance in patients undergoing therapy. The absence of Q47H mutations in majority of the study participants in our study implies that they will not respond well to daprevir and mericitabine. Screening of patients for pre-existing resistant mutations before commencement of therapy and monitoring during and after therapy are recommended.

Glucose controls lipolysis through Golgi PtdIns4P-mediated regulation of ATGL

Metabolic crosstalk of the major nutrients glucose, amino acids and fatty acids (FAs) ensures systemic metabolic homeostasis. The coordination between the supply of glucose and FAs to meet various physiological demands is especially important as improper nutrient levels lead to metabolic disorders, such as diabetes and metabolic dysfunction-associated steatohepatitis (MASH). In response to the oscillations in blood glucose levels, lipolysis is thought to be mainly regulated hormonally to control FA liberation from lipid droplets by insulin, catecholamine and glucagon. However, whether general cell-intrinsic mechanisms exist to directly modulate lipolysis via glucose sensing remains largely unknown. Here we report the identification of such an intrinsic mechanism, which involves Golgi PtdIns4P-mediated regulation of adipose triglyceride lipase (ATGL)-driven lipolysis via intracellular glucose sensing. Mechanistically, depletion of intracellular glucose results in lower Golgi PtdIns4P levels, and thus reduced assembly of the E3 ligase complex CUL7FBXW8 in the Golgi apparatus. Decreased levels of the E3 ligase complex lead to reduced polyubiquitylation of ATGL in the Golgi and enhancement of ATGL-driven lipolysis. This cell-intrinsic mechanism regulates both the pool of intracellular FAs and their extracellular release to meet physiological demands during fasting and glucose deprivation. Moreover, genetic and pharmacological manipulation of the Golgi PtdIns4P–CUL7FBXW8–ATGL axis in mouse models of simple hepatic steatosis and MASH, as well as during ex vivo perfusion of a human steatotic liver graft leads to the amelioration of steatosis, suggesting that this pathway might be a promising target for metabolic dysfunction-associated steatotic liver disease and possibly MASH.

Low Nitrogen Input Mitigates Quantitative but Not Qualitative Reconfiguration of Leaf Primary Metabolism in Brassica napus L. Subjected to Drought and Rehydration.

In the context of climate change and the reduction of mineral nitrogen (N) inputs applied to the field, winter oilseed rape (WOSR) will have to cope with low-N conditions combined with water limitation periods. Since these stresses can significantly reduce seed yield and seed quality, maintaining WOSR productivity under a wide range of growth conditions represents a major goal for crop improvement. N metabolism plays a pivotal role during the metabolic acclimation to drought in Brassica species by supporting the accumulation of osmoprotective compounds and the source-to-sink remobilization of nutrients. Thus, N deficiency could have detrimental effects on the acclimation of WOSR to drought. Here, we took advantage of a previously established experiment to evaluate the metabolic acclimation of WOSR during 14 days of drought, followed by 8 days of rehydration under high- or low-N fertilization regimes. For this purpose, we selected three leaf ranks exhibiting contrasted sink/source status to perform absolute quantification of plant central metabolites. Besides the well-described accumulation of proline, we observed contrasted accumulations of some "respiratory" amino acids (branched-chain amino acids, lysineand tyrosine) in response to drought under high- and low-N conditions. Drought also induced an increase in sucrose content in sink leaves combined with a decrease in source leaves. N deficiency strongly decreased the levels of major amino acids and subsequently the metabolic response to drought. The drought-rehydration sequence identified proline, phenylalanine, and tryptophan as valuable metabolic indicators of WOSR water status for sink leaves. The results were discussed with respect to the metabolic origin of sucrose and some amino acids in sink leaves and the impact of drought on source-to-sink remobilization processes depending on N nutrition status. Overall, this study identified major metabolic signatures reflecting a similar response of oilseed rape to drought under low- and high-N conditions.

Abstract 3067: SLC7A5/LAT1 promotes the Warburg Effect for TNBC progression through Trp/QPRT/NAD+ pathway

Abstract SLC7A5/LAT1 is a cross-membrane transporter that functions in uptake of large neutral amino acids into cells. Many pathological studies have shown that LAT1 is highly expressed in cancer tissue from various origins such as lung, biliary tract, breast, prostate, bone, and their metastatic legions. High LAT1 expression has been recognized as a significant prognostic marker in various cancers. Our TCGA data analysis showed that the expression level of LAT1 is higher in TNBC tumors compared with normal breast tissues. The higher expression level of LAT1 is linked with poor overall and progression-free survival. Dysregulation of amino acid transporters lead to metabolic reprogramming, which changes intracellular amino acid levels, contributing to the pathogenesis of cancer. However, it remains elusive whether and how LAT1 re-wires the cellular metabolic programs to promote TNBC tumor proliferation. In this study, we showed that inhibition of SLC7A5/LAT1 significantly reduced TNBC cell proliferation, viability, migration capability while promoting apoptosis. Mechanistically, we found significant positive correlations of LAT1 with PKM2 and LDHA in breast cancer tissues, which are key enzymes in mediating glycolysis and lactate fermentation or the Warburg Effect. Knock down LAT1 via siRNA decreased the expression levels of p-PKM2 and p-LDHA in TNBC cells. Tryptophan, a major substrate amino acid transported into cytosol by LAT1, is the precursor for the de novo synthesis of nicotinamide adenine dinucleotide (NAD+). The enzyme quinolinic acid phosphoribosyltransferase (QPRT) catalyzes the conversion from quinolinate, a product of tryptophan degradation, to NAD+. We showed LAT1 inhibition reduced the expression level of QPRT, cytosolic NAD+/NADH ratio as well as lactate production. Addition of L-Tryptophan increased the levels of p-PKM2 and p-LDHA while knockdown of LAT1 reversed tryptophan-mediated increase of PKM2 and LDHA activities, suggesting that LAT1 may regulate PKM2/LDHA through Trp/QPRT/NAD+ pathway for promoting the Warburg Effect. Additionally, we found upregulations of SLC7A5/LAT1, p-PKM2 and p-LDHA in doxorubicin-resistant cells and PDX mouse model compared with their corresponding controls. Collectively, these results provide evidence that overexpression of LAT1 in TNBC drives cancer progression and drug resistance by reinforcing the Warburg Effect, suggesting LAT1 as a promising therapeutic target for addressing TNBC. Our ongoing work will evaluate whether inhibiting LAT1 genetically or pharmacologically reduces xenograft tumor growth and enhances the sensitivities of TNBC resistant cells to chemotherapy in vivo. Our investigation will elucidate the underlying mechanisms involved in these processes. Citation Format: Margot Y. Fedoroff, Lei Zhao, Leili Saeednejad Zanjani, Jun He. SLC7A5/LAT1 promotes the Warburg Effect for TNBC progression through Trp/QPRT/NAD+ pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3067.

Neurotoxicity and brain metabolic dysfunction induced by long-term food-derived arsenic exposure

Rice consumption are a major cause of long-term exposure to arsenic. However, the exact mechanism of the neurotoxic action caused by arsenic exposure remain elusive. The purpose of this study was to investigate the effects of arsenic exposure on the behavior and neuronal structure of Caenorhabditis elegans, followed by the establishment of a 14-month mice model to simulate long-term effects. To elucidate the toxic effects of arsenic on brain metabolism, comprehensive analyses of neurotransmitter content and brain metabolomics were conducted. The findings demonstrated that arsenic exposure led to behavioral impairment, reduced levels of glutamatergic, GABAergic, dopaminergic and serotonergic neurotransmitters, as well as structural abnormalities. The majority of amino acid and fatty acid compounds were found to be upregulated in the hippocampus and cortical tissues. Respectively, an increase in nucleic acid and hydroxy acid-related compounds was observed in hippocampal tissues, while an upregulation of purine ribose-related compounds was detected in cortical tissues. The identified differential metabolites were primarily impacted by glutathione metabolism, arginine and proline metabolism, as well as alanine, aspartate, and glutamate metabolism. Correlation analysis of the hippocampus and cortex yielded compelling evidence of a strong association between glutamate and the majority of the presented metabolites.

Novel Aquareovirus isolated from channel catfish (Ictalurus punctatus) used in mussel restoration efforts in Wisconsin.

Channel catfish (Ictalurus punctatus) are a food fish extensively reared in aquaculture facilities throughout the world and are also among the most abundant wild catfish species in North America, making them a popular target of anglers. Furthermore, channel catfish are important members of aquatic ecosystems; for example, they serve as a glochidial host for the endangered winged mapleleaf mussel (Quadrula fragosa), making them critical for conserving this species through hatchery-based restoration efforts. During a routine health inspection, a novel aquareovirus was isolated from channel catfish used in mussel propagation efforts at a fish hatchery in Wisconsin. This virus was isolated on brown bullhead cells (ATCC CCL-59) and identified through metagenomic sequencing as a novel member of the family Spinareoviridae, genus Aquareovirus. The virus genome consists of 11 segments, as is typical of the aquareoviruses, with phylogenetic relationships based on RNA-dependent RNA polymerase and major outer capsid protein amino acid sequences showing it to be most closely related to golden shiner virus (aquareovirus C) and aquareovirus C/American grass carp reovirus (aquareovirus G) respectively. The potential of the new virus, which we name genictpun virus 1 (GNIPV-1), to cause disease in channel catfish or other species remains unknown.

Silica Nanoparticles Decrease Glutamate Uptake in Blood–Brain Barrier Components

Glutamate is the major excitatory amino acid in the vertebrate brain, playing an important role in most brain functions. It exerts its activity through plasma membrane receptors and transporters, expressed both in neurons and glia cells. Overstimulation of neuronal glutamate receptors is linked to cell death in a process known as excitotoxicity, that is prevented by the efficient removal of the neurotransmitter through glutamate transporters enriched in the glia plasma membrane and in the components of the blood–brain barrier (BBB). Silica nanoparticles (SiO2-NPs) have been widely used in biomedical applications and directed to enter the circulatory system; however, little is known about the potential adverse effects of SiO2-NPs exposure on the BBB transport systems that support the critical isolation function between the central nervous system (CNS) and the peripheral circulation. In this contribution, we investigated the plausible SiO2-NPs-mediated disruption of the glutamate transport system expressed by BBB cell components. First, we evaluated the cytotoxic effect of SiO2-NPs on human brain endothelial (HBEC) and Uppsala 87 Malignant glioma (U-87MG) cell lines. Transport kinetics were evaluated, and the exposure effect of SiO2-NPs on glutamate transport activity was determined in both cell lines. Exposure of the cells to different SiO2-NP concentrations (0.4, 4.8, 10, and 20 µg/ml) and time periods (3 and 6 h) did not affect cell viability. We found that the radio-labeled D-aspartate ([3H]-D-Asp) uptake is mostly sodium-dependent, and downregulated by its own substrate (glutamate). Furthermore, SiO2-NPs exposure on endothelial and astrocytes decreases [3H]-D-Asp uptake in a dose-dependent manner. Interestingly, a decrease in the transporter catalytic efficiency, probably linked to a diminution in the affinity of the transporter, was detected upon SiO2-NPs. These results favor the notion that exposure to SiO2-NPs could disrupt BBB function and by these means shed some light into our understanding of the deleterious effects of air pollution on the CNS.

Dissection of the spatial dynamics of biosynthesis, transport, and turnover of major amino acids in tea plants (Camellia sinensis)

Abstract High levels of free amino acids (AAs) in tea leaves are crucial for tea flavor and health function, however, the dynamic AA biosynthesis, transport and turnover in tea plants remain elusive. Here we dissected whole tea plants for these dynamics by assessing AA profiles and transcriptomes of metabolic pathway genes in tea roots, stems, and leaves and revealing their distinctive features with regard to AA synthesis, transport, and degradation/recycling. Nitrogen assimilation dominated in the roots wherein glutamine (Gln), theanine, and Arginine (Arg) were actively synthesized. Arg was transported into trunk roots and stems, together with Glu, Gln, and theanine as the major AAs in the xylem sap for long-distance root-to-leaf transport. Transcriptome analysis revealed that genes involved in Arg synthesis were highly expressed in roots, but those for Arg transport and degradation were highly expressed in stems and young leaves, respectively. CsGSIa transcripts were found in root meristem cells, root, stem and leaf vascular tissues, and leaf mesophyll where it appeared to participate in AA synthesis, transport and recycling. Overexpression CsGSIa in tea transgenic hairy roots and knockdown of CsGSIa in transgenic hairy roots and tea leaves produced higher and lower Gln and theanine than wild type roots and leaves, respectively. This study provides comprehensive and new insights into AA metabolism and transport in the whole tea plant.

Ameliorative Effect of Macadamia Nut Protein Peptides on Acetaminophen-Induced Acute Liver Injury in Mice.

This study aims to examine the ameliorative effect of macadamia nut protein peptides (MPP) on acetaminophen (APAP)-induced liver injury (AILI) in mice, and develop a new strategy for identifying hepatoprotective functional foods. The molecular weight distribution and amino acid composition of MPP were first studied. Forty mice were then randomized into four groups: control group (CON), APAP model group, APAP+MPP low-dose group (APAP+L-MPP), and APAP+MPP high-dose group (APAP+H-MPP). The APAP+L-MPP (320 mg/kg per day) and APAP+H-MPP (640 mg/kg per day) groups received continuous MPP gavage for 2 weeks. A 12 h of APAP (200 mg/kg) gavage resulted in liver damage. Pathological alterations, antioxidant index levels, expression of toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB), and associated inflammatory factors were determined for each treatment group. The results revealed that the total amino acid content of MPP was 39.58 g/100 g, with Glu, Arg, Asp, Leu, Tyr, and Gly being the major amino acids. The molecular weight range of 0-1000 Da accounted for 73.54%, and 0-500 Da accounted for 62.84% of MPP. MPP ameliorated the pathological morphology and reduced the serum levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase of AILI in mice. MPP significantly increased the activities of superoxide dismutase and glutathione peroxidase in the liver compared with the APAP group. MPP inhibited the expression of TLR4, NF-κB, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) genes in AILI mice. MPP also inhibited the expression levels of inflammatory factors (TNF-α and IL-6). Our study concludes that MPP alleviates AILI in mice by enhancing antioxidant capacity and inhibiting TLR4/NF-κB pathway-related gene activation.

Changes and correlation analysis of volatile flavor compounds, amino acids, and soluble sugars in durian during different drying processes

Durian contains rich flavor components that undergo complex changes during drying. In this study, durian was subjected to integrated freeze-drying (IFD), conventional freeze-drying (CFD), and hot air drying (AD). Compared with the fresh samples, those dried by IFD, CFD, and AD lost 11, 9, and 7 original volatile compounds, respectively, and generated 7, 6, and 8 new volatile compounds, respectively, and showed a rapid and then slow decreasing trend in the total content during drying. However, the types of amino acids and soluble sugars remained unchanged during each of the drying methods. Furthermore, volatile compounds showed a significant negative correlation with the majority of amino acids and a significant positive correlation with soluble sugars. The IFD samples had the highest content of volatile compounds, amino acids, and soluble sugars. Therefore, IFD is recommended as a preferable drying method for durian.

Proximate composition, physico-chemical properties, techno-functional properties, nutritional quality, and functional activity of Ferula assafoetida oleo-gum-resin

Mass and tear assafoetida are mainly composed of carbohydrates (61.14 and 61.50 g/100 g) followed by protein (13.45 and 14.28 g/100 g), fat (11.85 and 11.50 g/100 g), and essential oil (9.25 and 8.40 g/100 g). Also measured were electrical conductivity (1250 and 1237 microsiemens/cm), osmolarity (192 and 198 milliosmol/kg), surface tension (38 and 41 mN/m), zeta potential (–23.7 and –23.5 mV), particle size (265 and 275 nm), particle size distribution (0.73 and 0.93), and viscosity (1.40 and 1.35 mPa.s), along with water content (12.80 and 11.25%), water solubility (65.68 and 72.50%), water swelling (145.8 and 137.0%), hygroscopicity (39.84 and 41.20%), hydrophobicity (7.30 and 6.85 µg/g), emulsification activity (72.66 and 70.0%), emulsification stability (66.97 and 64.0%), foam activity (55.09 and 58.50%), foam stability (48.95 and 52.0%), water-holding capacity (3.86 and4.20 g/g) and oil-holding capacity (3.35 and 3.15 g/g). The major amino acids in assafoetida proteins were isoleucine, methionine, glycine, aspartic acid, alanine, glutamic acid, and lysine. The main fatty acids of assafoetida are myristic, palmitic, alpha-linolenic, oleic, linoleic, and lauric acids. The main volatile oil components of assafoetida are 1-propenyl butyl disulfide, α-pinene, bis (1-methyl thio)propyl disulfide, and 1-(propylthio)propyl propyl disulfide. Assafoetida showed good antioxidant capacity, anti-amylase, and anti-glucosidase activity similar to positive controls.

Preparação de farinha de resíduos de tilápia do Nilo (Oreochromis niloticus) para consumo humano

ABSTRACT: This study aimed to prepare and characterize Nile tilapia (Oreochromis niloticus) waste meal. The experimental design consisted of three treatments (carcass meal, mechanically separated meat meal - MSM, and head meal) and five replications of each treatment. Yield, microbiological quality, pH, water activity, proximate composition, energy value, mineral composition, fatty acid profile, amino acid composition, color, and particle size fractions and the geometric mean particle diameter (GMD) analyses were performed. Data were subjected to ANOVA, and means were compared by Tukey’s test at P < 0.05. Carcass, mechanically separated meat, and head meals had a protein content of 56.45%, 78.60%, and 50.33%, respectively, a lipid content of 7.16%, 13.15%, and 4.58%, respectively, and an ash content of 32.61%, 4.26%, and 38.41%, respectively. The mean yield of fish waste meals was 13%. Oleic, palmitic, and linoleic acids were the major fatty acids in the three fish waste meals, and glutamic acid, leucine, lysine, and glycine were the major amino acids. Color analysis showed that lightness (L*) ranged from 55.32 to 77.19, redness (a*) from 4.44 to 4.88, and yellowness (b*) from 4.13 to 9.51. The GMD ranged from 0.37 to 0.99 mm. The results indicate that Nile tilapia waste meals have high nutritional value and adequate pH, water activity, and microbiological quality for use in human nutrition.

Screening of phytoconstituents from Bacopa monnieri (L.) Pennell and Mucuna pruriens (L.) DC. to identify potential inhibitors against Cerebroside sulfotransferase.

Cerebroside sulfotransferase (CST) is considered a target protein in developing substrate reduction therapy for metachromatic leukodystrophy. This study employed a multistep virtual screening approach for getting a specific and potent inhibitor against CST from 35 phytoconstituents of Bacopa monnieri (L.) Pennell and 31 phytoconstituents of Mucuna pruriens (L.) DC. from the IMPPAT 2.0 database. Using a binding score cutoff of -8.0 kcal/mol with ADME and toxicity screening, four phytoconstituents IMPHY009537 (Stigmastenol), IMPHY004141 (alpha-Amyrenyl acetate), IMPHY014836 (beta-Sitosterol), and IMPHY001534 (jujubogenin) were considered for in-depth analysis. In the binding pocket of CST, the major amino acid residues that decide the orientation and interaction of compounds are Lys85, His84, His141, Phe170, Tyr176, and Phe177. The molecular dynamics simulation with a 100ns time span further validated the stability and rigidity of the docked complexes of the four hits by exploring the structural deviation and compactness, hydrogen bond interaction, solvent accessible surface area, principal component analysis, and free energy landscape analysis. Stigmastenol from Bacopa monnieri with no potential cross targets was found to be the most potent and selective CST inhibitor followed by alpha-Amyrenyl acetate from Mucuna pruriens as the second-best performing inhibitor against CST. Our computational drug screening approach may contribute to the development of oral drugs against metachromatic leukodystrophy.

Effect of choline amino acid ionic liquids on maize seed germination and endogenous plant hormone levels.

Prior research has established choline-based ionic liquids (ILs) as safe for various organisms. However, their impact on plants has been underexplored. To identify effective eco-friendly ILs, we synthesized seven choline amino acid ([Chl][AA]) ILs and analyzed their physiological influence on maize seed germination. In contrast to the traditionally used N-octyl pyridinium bromide IL, these seven [Chl][AA] ILs exhibited substantially lower toxicity. Moreover, within a broad treatment concentration range (10-100 mg L-1), these ILs notably enhanced maize germination indices and root and shoot growth. Specifically, treatment with 100 mg L-1 choline tryptophan resulted in a 21.2% increase in germination index compared to those of control maize. Compared to the control, the application of choline serine, choline aspartic acid, choline phenylalanine, and choline tryptophan at 100 mg L-1 led to respective increases of 23.9%, 21.5%, 22.5%, and 24.5% in maize shoot length. Analysis of endogenous hormones and free amino acid contents revealed elevated levels of growth-promoting plant hormones (gibberellic acid and zeatin) in maize shoot tips, as well as increased contents of major amino acids (glutamate, glycine, and arginine) following treatment with different [Chl][AA] ILs at 100 mg L-1. These findings indicate that [Chl][AA] holds promise for the development and application of novel low-toxicity ILs.