Search Results (319)

Japonica rice offers high cost-effectiveness and yield, with the potential to replace glutinous rice in Chinese sweet rice wine (CSRW) brewing. It can be classified into aromatic and non-aromatic types, but whether different varieties cause flavor and metabolite differences in CSRW remains unclear. In this study, glutinous rice (GR), three aromatic japonica varieties (CS-217, HXR-450, SXJ-1018), and two non-aromatic varieties (TA-1, HR-1212) were used as raw materials. The qualities of different CSRWs were evaluated through physicochemical indices, sensory evaluation, phenolic and flavonoid contents, antioxidant capacities, HS-GC-IMS, and UPLC-MS/MS. The results showed that CS-217 displayed the highest total acid content, along with excellent overall sensory evaluation, total phenolic content, and antioxidant capacity. A total of 28 VOCs were identified by HS-GC-IMS, among which 13 compounds with VIP ≥ 1, including butyl isobutyrate, butyl acetate, and ethyl pentanoate, were identified as key flavor discriminant factors. Additionally, 2501 non-volatile metabolites were identified, and five key metabolic pathways were revealed. These pathways synergistically regulate CSRW flavor and nutritional quality. Different japonica rice varieties exhibited respective advantages in CSRW quality indicators, providing a basis for the diversification of raw materials in CSRW production. Full article

Pyrrolidinyl-substituted silacyclopentane (CH₂)₄Si(Pyr)₂ and α-amino isobutyric acid (H₂Aib) react with the release of one equivalent pyrrolidine (HPyr) and the formation of the pentacoordinate silicon bis-chelate (Aib)Si(CH₂)₄(HPyr), which features the di-anion of the amino acid as an (O,N)-chelator and one equivalent of pyrrolidine as an additional lone-pair donor. Crystallographic analyses of the chloroform solvate (Aib)Si(CH₂)₄(HPyr)·(CHCl₃), which undergoes a phase transition at 200 K, and a solvent-free modification (Aib)Si(CH₂)₄(HPyr), which features two crystallographically independent molecules of the complex, revealed that the N atom of the HPyr ligand, as well as the carboxylate of Aib, occupy the axial positions in the trigonal bipyramidal Si coordination sphere; the Si–C bonds of the silacyclopentane rest on equatorial sites. For the isolated molecule in a solvent environment, computational analyses revealed that the energy difference between this configuration and the related isomer with an equatorial HPyr and equatorial–axial positioning of the silacyclopentane motif is marginal. In DMSO solution, the adduct (Aib)Si(CH₂)₄(HPyr) decomposed, forming the hexacoordinate Si complex (HAib)₂Si(CH₂)₄ as one of the decomposition products. In a deliberate manner, this compound was accessible from the diethylamino-substituted silacyclopentane (CH₂)₄Si(NEt₂)₂ and H₂Aib with the liberation of diethylamine. (HAib)₂Si(CH₂)₄ features two mono-anions of the α-amino acid as (O,N)-chelators, their carboxylate O atoms are trans-disposed to silacyclopentane, and their NH₂ groups are mutually trans. Full article

Dietary supplementation with sodium butyrate or bovine colostrum modulates the gut–liver axis in weaned piglets. Sodium butyrate exerted beneficial effects on liver function and lipid parameters, while also inhibiting inflammation and promoting the maintenance of the intestinal barrier. A particularly pronounced effect was observed with bovine colostrum supplementation, which significantly increased average daily weight gain (p < 0.001). In addition, piglets receiving colostrum consumed more feed and exhibited a significantly lower feed conversion ratio (p = 0.002). Metabolic changes induced by sodium butyrate and bovine colostrum supplementation resulted in alterations in the hepatic fatty acid profile, including a reduction in n-3 polyunsaturated fatty acids and a decrease in collagen fiber content in the liver (p = 0.03). The nutritional interventions did not significantly affect microbial diversity indices; however, marked changes in volatile fatty acid concentrations were observed in the large intestine. These changes indicate enhanced microbial fermentation and increased nutrient absorption in the experimental groups. Significant increases were detected in acetic acid (p = 0.003) as well as in butyric, isobutyric, and valeric acids (p = 0.014, p = 0.024, and p = 0.038, respectively). Supplementation with sodium butyrate and dried bovine colostrum also led to increased hepatic concentrations of macro- and microelements in piglets from the experimental groups. Genomic analyses suggest that sodium butyrate modulates hepatic metabolic and inflammatory pathways by downregulating PPAR (peroxisome proliferator-activated receptor) and SIRT3 (sirtuin 3) expression and reducing TNF (tumor necrosis factor) gene expression, highlighting its potential role in regulating lipid metabolism, oxidative stress, and inflammation in a porcine model. Overall, the results indicate that both supplements may contribute to the modulation of gut microbial activity and liver metabolism in weaned piglets. Full article

Background: Ketamine is a rapid-acting antidepressant for major depressive disorder; however, its effects are short-lasting and associated with neurotoxic side effects. Thus, identifying strategies to prolong its antidepressant effects is of critical importance. It has been shown that Dajianzhong Decoction (DJZT) prolongs the antidepressant effects of ketamine through modulation of the gut microbiota, but the underlying mechanisms remain unclear. Method: Fecal microbiota transplantation, metabolomic profiling, pharmacological interventions, and behavioral approaches were employed together with a chronic unpredictable mild stress (CUMS) mouse model to investigate how microbiota-derived signals mediate the combined effects of DJZT and ketamine. Results: Microbiota from CUMS mice induced depressive-like behaviors in recipient mice, accompanied by reduced levels of short-chain fatty acids (SCFAs), decreased FFAR2 expression in the medial prefrontal cortex, and increased neuroinflammation and synaptic deficits. These alterations were reversed by microbiota from DJZT-plus-ketamine-treated donors. Notably, acetic acid and isobutyric acid were identified as key SCFAs restored by the combined treatment and were significantly associated with behavioral outcomes. Moreover, SCFA supplementation recapitulated these effects by activating FFAR2 and suppressing NLRP3–IL-1β signaling. Importantly, pharmacological inhibition of FFAR2 using GLPG0974 abolished the antidepressant-like, anti-inflammatory, and synaptic protective effects of the microbiota from DJZT-plus-ketamine-treated donors. Conclusions: These findings demonstrate that microbiota-derived SCFAs mediate the synergistic antidepressant effects of DJZT and ketamine via a central FFAR2-dependent mechanism involving suppression of neuroinflammation. This work highlights a potential role of the SCFA–FFAR2–NLRP3– IL-1β axis in influencing ketamine efficacy and points to microbiota-modulating strategies as a possible avenue for improving antidepressant therapy. Full article

The bean bug Riptortus pedestris is one of the principal soybean pests throughout East Asia. Males release an aggregation pheromone consisting of three compounds—(E)-2-hexenyl (Z)-3-hexenoate (E2HZ3H), (E)-2-hexenyl (E)-2-hexenoate (E2HE2H) and myristyl isobutyrate (MI)—in a ratio of 1:5:1. Although pheromone-based baits have been extensively deployed to control the pest for the past three decades, the glands responsible for producing these volatiles have remained unidentified. Our investigations revealed that the metathoracic glands (MTGs) of males synthesized the two pheromonal esters, E2HZ3H and E2HE2H, along with a suite of auxiliary compounds: (E)-2-hexenal, hexanoic acid, (E)-3-hexenoic acid, (E)-2-hexenoic acid, (E)-2-octenal, and (E)-2-hexenyl hexanoate (E2HH). Female MTGs were morphologically similar to those of males and generated the same auxiliary compounds, but lacked the capacity to produce the pheromonal esters E2HZ3H and E2HE2H. MI was released exclusively from the male abdominal sternites. Its quantity was nearly uniform across all sternal segments, implying secretion from an epidermal glandular patch (GP) that was evenly distributed over the sternites. In nymphs, dorsal abdominal glands (DAGs) emitted several volatiles—including (E)-2-hexenal, 4-oxo-(E)-2-hexenal, (E)-2-octenal, and (E)-2-octenoic acid—some of which are characteristic defensive compounds of heteropterans. Thus, our study clarifies how sex- and stage-specific glands contribute to the chemical polymorphism observed in R. pedestris. Full article

Consumers currently consider organic foods superior to conventional ones. They regard them as more environmentally friendly and healthier. The sensory and volatile properties, as well as the antioxidant content of the Italian organic almond of the “Tuono” cultivar, were evaluated in this study. The following methods were used: sensory analysis, determination of total antioxidant capacity and the HS-SPME sampling technique followed by GC/MS analysis for the analysis of volatile compounds. Our findings highlighted the enhanced sensory quality of the organic sample in comparison to the conventional one. The presence of almond aroma, marzipan/benzaldehyde, tobacco, floral notes, sweetness, and crunchiness was exhibited by the analysed organic samples. The floral attribute is especially prominent, with its concentration being roughly four times higher in organic almonds than in conventional ones (4.96 vs. 1.25). There was no statistically significant difference in total phenolic content and antioxidant capacity between organic and conventional almonds. Significant differences were found between the organic and conventional systems for the volatile profile. Organic almonds were characterised by a higher presence of butanol in comparison to conventional (5.2 vs. 1.3, respectively) and limonene (3 vs. 1.5, respectively), both of which are associated with fruity aromas. Higher levels (expressed as %) of 2-methylbutanal, 3-methylbutanal, isobutyric acid, 2-heptanone, 3-heptanone, octanoic acid, and pinacol were also found in organic almonds. The possibility of producing almonds of superior sensory quality through organic systems could be considered a key factor in the potential contribution to maintaining the sustainability of agroecosystems. Full article

This study evaluated the effects of dietary PUFA supplementation on growth performance, serum biochemical indices, rumen morphology, ruminal fermentation, and rumen microbial communities in Hu sheep. Thirty healthy male Hu sheep (80 days old; 18.70 ± 0.72 kg) were randomly assigned to three groups (n = 10/group) and fed a basal diet supplemented with 4% rumen-bypass palmitic acid fat powder (POS), 4% linseed oil (LO; rich in ω-3 PUFA), or 4% sunflower oil (SO; rich in ω-6 PUFA). PUFA supplementation did not affect average daily gain or rumen tissue morphology (p > 0.05), but it significantly reduced serum creatinine, uric acid, and high-density lipoprotein concentrations (p < 0.05). Orthogonal comparative analysis showed that supplementing with PUFA significantly reduced acetate and propionate, while increasing isobutyrate, butyrate, isovalerate, and TVFAs (p < 0.05). The levels of propionate, butyrate and TVFAs in the SO group were higher than those in the LO group (p < 0.05), and the ratio of acetate to propionate was lower (p < 0.05). Supplementing with PUFA reshaped the rumen microbiota, increasing the relative abundances of Bacteroidota, Firmicutes, Euryarchaeota, Cyanobacteria, and Actinobacteriota, while decreasing Proteobacteria and Desulfobacterota (p < 0.05). At the genus level, Prevotella and Rikenellaceae_RC9_gut_group were enriched in both groups with added PUFA, while Prevotellace-UCG-001 was specifically enriched in the LO group; in contrast, Prevotella_7, Succinivibriaceae_UCG-001, Prevotella_9, and Dialist all showed a decrease (p < 0.05). The alpha diversity increased, while the beta diversity showed significant differences between the group with added PUFA and the control group. Functionally, LO enriches carbohydrate and energy metabolism, while SO enriches nucleotide metabolism. In summary, linseed oil and sunflower oil improved serum indices and ruminal fermentation without impairing growth, but through distinct microbial and functional pathways. Full article

A study was conducted to evaluate the effect of neutral detergent fiber (NDF) levels of calf starter and weaning time on growth, rumen fermentation characteristics, serum metabolites, and rumen microbial diversity of Holstein calves. A total of 24 newly born male Holstein calves were randomly distributed to four groups in a completely randomized design with a 2 × 2 factorial arrangement of NDF levels (14% and 24%) and weaning time (d 44 and d 54). There was no interaction between starter NDF levels and weaning time for any trait except rumen acetic acid in the immediate post-weaning phase (p = 0.013). Starter NDF levels had no effect on growth, feed intake, and hay intake. Late-weaned calves had greater (p = 0.050) weight gain in the pre-weaning phase whereas, early-weaned calves showed greater weight gain (p = 0.004) and starter intake (p = 0.004) in the post-weaning phase although overall weight gain, and starter and hay intakes were not affected by weaning time. Rumen pH, ammonia nitrogen, and most volatile fatty acids remained unaffected by starter NDF levels and weaning except isobutyric acid which was greater in calves fed 24% NDF starter (p = 0.001) in the immediate post-weaning and isovaleric acid which was greater in early-weaned calves (p = 0.044) at the end of experiment. Serum metabolites were largely affected (p < 0.05) by starter NDF levels and weaning time in the pre-weaning phase only. Alpha diversity of rumen microbes was greater and chaotic in 14% NDF starter group (early- and late-weaned) in the pre-weaning phase which converged in the immediate post-weaning phase and diverged on starter NDF basis at the end of experiment. Microbial ecology at phylum and genus levels composition were greatly driven by starter NDF levels in the pre-weaning phase, by weaning time in the immediate post-weaning phase, and two distinct bifurcated microbial ecologies based on starter NDF levels appeared at the end of experiment. In conclusion, the comparable growth with distinct microbial diversity but largely in favor of 24% NDF starter suggests that calves can be subjected to early weaning with 24% starter NDF levels for smooth transition from liquid to solid feed in Holstein calves. Full article

Akkermansia muciniphila, a next-generation probiotic in the human intestinal mucus layer, exhibits significant health-promoting properties. However, traditional static culture systems fail to replicate the dynamic peristaltic environment of the gastrointestinal tract, limiting understanding of its metabolic characteristics. This study employed an improved gastrointestinal bioreactor simulating intestinal peristalsis to investigate A. muciniphila growth dynamics and metabolomic profiles under dynamic conditions. Dynamic cultivation significantly enhanced bacterial growth. Biomass reached 1.32 ± 0.03 g/L in bovine heart infusion (BHI) medium and 2.03 ± 0.05 g/L in BHI supplemented with 2.5 g/L porcine mucin. These values represent increases of 45.05% and 123.08% relative to static BHI cultures, respectively. Dynamic conditions markedly elevated short-chain fatty acid production (acetic, propionic, isobutyric, isovaleric acids). Untargeted metabolomics identified 1463 metabolites with 1294 showing significant differential expression. Dynamic cultivation substantially altered amino acid biosynthesis, fatty acid, purine, and pyrimidine metabolism. These findings advance the understanding of A. muciniphila physiology and provide insights into its metabolic characteristics under simulated intestinal conditions. Full article

Background/Objectives: Recently, a randomized clinical trial evaluated whether a six-month probiotic administration could reduce symptom severity in preschool children with Autism Spectrum Disorders (ASD), with (GI) or without (NGI) gastrointestinal symptoms. Significant positive changes were observed only in NGI children. A second explorative study on children prior to intervention identified a fecal metabolome fingerprint associated with ASD severity. Building on these findings, the present study aimed to assess whether metabolomics could monitor changes in ASD severity following probiotic administration using a subset of samples from the same trial. Second, this study aimed to identify fecal metabolites to be monitored in children to predict whether their autism severity may decrease after probiotic or placebo treatment. Methods: Evaluations of the fecal metabolome and microbiota could be completed on 57 children before and after a double-blind administration of a probiotic mixture or a placebo. Results: In NGI children the probiotic was found to influence the concentration of the amino acids aspartate, leucine, tryptophan, and valine, together with nicotinate and the short chain fatty acids acetate, butyrate, isobutyrate, and propionate. Lactobacilli and Sutterella showed significant changes in response to probiotic administration (p < 0.05). Acetate, 4-hydroxyphenyl, galactose, proline, and tyramine were identified as key fecal metabolites for prediction purposes. Conclusions: The present exploratory analysis, despite the small sample size, suggests that fecal metabolomics may provide a useful approach for monitoring and potentially for predicting changes in ASD severity following probiotics administration. Full article

Styela plicata, an edible ascidian rich in diverse bioactive constituents, represents a promising source of marine natural products for therapeutic discovery. Here, bioactive components from a 95% ethanol extract of S. plicata (ESP) were characterized by HPLC-MS/MS, showing that the major constituents were oxygenated small molecules dominated by fatty acyls and carboxylic acid derivatives. In a mouse model of alcohol-induced liver injury, H-ESP treatment (300 mg/kg) significantly reduced serum levels of AST, ALT, and TG (p < 0.01), while effectively ameliorating pathological changes in liver tissue, reducing lipid accumulation and inflammatory responses. Transcriptome sequencing (H-ESP vs. model group) identified 1097 differentially expressed genes (172 upregulated and 925 downregulated), and KEGG analysis highlighted significant enrichment of the Toll-like receptor signaling pathway. ESP modulated hepatic metabolite expression, suppressed inflammation via TLR-4/NF-κB pathway inhibition, and boosted antioxidant defenses by activating Nrf2/HO-1 signaling, which was further confirmed by RT-qPCR and immunohistochemistry. ESP increased intestinal SCFAs (acetate, propionate, isobutyrate; p < 0.05), improved α-diversity and the Firmicutes/Bacteroidetes ratio, reversed shifts in Lactobacillus and Bifidobacterium, and partly restored Odoribacter, supporting a gut–liver axis mechanism. Overall, these findings indicate that ESP exerts hepatoprotective effects by modulating the gut–liver axis, and they provide insights for developing natural therapeutics against alcoholic liver disease. Full article

Peptaibols, predominantly secreted by Trichoderma species, are a class of linear peptides composed of five to twenty amino acid residues, synthesized non-ribosomally and enriched with α-amino isobutyric acid. These unique peptides appear to be highly effective in mediating the interactions between Trichoderma and plant pathogenic fungi. In this study, Ultra-Performance Liquid Chromatography–Quadrupole Time-Of-Flight Mass Spectrometry/Mass Spectrometry (UPLC-QTOF-MS/MS) technology was used to detect peptaibols profiles of Trichoderma strains during their interactions with the pathogen Fusarium graminearum. MS investigations of crude extracts derived from in vitro confrontations of Trichoderma atroviride T23 and its genetically modified counterparts, dual-culture assays of Mlae1, Mvel1, OElae1, and OEvel1 with F. graminearum were performed to shed light on the regulatory role of the velvet complex composed of LAE1&VEL1 in the synthesis of peptaibols during the microbial interaction. These results revealed intriguing variations in the total peptaibols produced during the interactions, as well as some differences in the specific peptaibol profiles between the confrontation and control tests. The overexpression strains, OElae1 and OEvel1, distinguished themselves by their proficiency in inducing long-residue peptaibols synthesis, attaining an impressive biocontrol index of up to 76%. The crude extracts containing peptaibols of OElae1 and OEvel1 demonstrated a capability to enhance cell membrane permeability and decrease DON toxin production in F. graminearum, and the crude extracts of OElae1 strains exhibited more effectiveness in reducing DON toxin production. In conclusion, the interaction with F. graminearum significantly impacted the peptaibol production in the examined Trichoderma strain, emphasizing the intricate interplay and reciprocal influence of genetic factors and environmental stimuli. Full article

Nigella sativa meal (NSM) is a by-product of the oil extraction process with potential use as a functional feed ingredient in ruminant nutrition due to its rich bioactive compounds and nutrient content. Therefore, this replicated (n = 6) completely randomized design (CRD) study aimed to firstly characterize NSM for its the bioactive compounds by chromatography–mass spectrometry (GC–MS) and liquid chromatography–MS (LC–MS). The effects of its dietary inclusion at 0%, 5%, 7.5%, and 10% NSM on in vitro rumen fermentation, gas production, CH₄ production, and rumen degradability were analyzed. Rumen fluid was collected orally from five sheep (body weight 20 ± 2 kg/head) and incubated with basal diets supplemented with pre-determined levels of NSM. The identified bioactive compounds identified included palmitic acid, oleic acid, linoleic acid, phenolic, flavonoid, thymoquinone, and saponin. The results showed that the dietary NSM supplementation in the diet significantly increased (p < 0.05) gas production at 18 and 48 h, NH₃–N concentration, IVDMD (In vitro dry matter degradability), and IVOMD (In vitro organic matter degradability). However, there was no significant effect (p > 0.05) on gas production at 12 and 24 h, and CH₄ production, pH, acetate, propionate, iso-butyrate, butyrate, iso-valerate, valerate, the acetate-to-propionate ratio (A:P), or total VFA concentration. Rumen fermentation was optimally modulated up to 10% without adverse effects on digestibility or CH₄ production. In this context, NSM acted as a functional feed ingredient in vitro. Therefore, in vivo analyses are required to confirm the efficacy under practical feeding conditions. Full article

Few studies have investigated the effects of L-valine (L-Val) supplementation on in vitro rumen fermentation parameters and methane (CH₄) production in low-nitrogen diets for ruminants. Therefore, we examined the impact of L-Val supplementation in low-protein diets on in vitro rumen fermentation parameters, CH₄ production, and microbial community structure. Two crude protein (CP) levels and 4 L-Val levels were tested as follows: CON group (Control group with 14.05% CP), LD group (low-nitrogen diets with 11.26% CP), LVA group (LD group + 0.25% L-Val), LVB group (LD group + 0.5% L-Val), LVC group (LD group + 0.75% L-Val), and LVD group (LD group + 1% L-Val). The experiment was conducted at five time points (2 h, 4 h, 8 h, 12 h, 24 h), with three replicates per treatment at each time point. Results indicated the following: (1) Fermentation pH decreased overall with time; at each time point, the LVB group exhibited the highest pH, significantly higher than the LD, LVC, and LVD groups (p < 0.05). (2) Ammonia nitrogen (NH₃-N) concentration increased over time, with LVA~LVD groups showing higher levels than the LD group at 24 h, while showing no difference compared to the CON group (p > 0.05). (3) Microbial protein (MCP) trends aligned with NH₃-N, with the LVB group exhibiting higher MCP than the LD group, while showing no difference compared to the CON group (p > 0.05). (4) Compared to the LD group, adding 0.5~1% L-Val increased acetic acid, total VFA (TVFA), and isobutyric acid concentrations at 4 h, 8 h, and 24 h fermentation (p < 0.05). (5) The LVB group exhibited higher proportions of protozoa and Fibrobacter succinogenes (F. succinogenes) compared to the LD group (p < 0.05). The proportion of F. succinogenes showed no significant difference from the CON group (p > 0.05), while the proportion of Butyrivibrio fibrisolvens (B. fibrisolvens) decreased when L-Val addition exceeded 0.5%. (6) Correlation analysis revealed positive correlation between protozoa and TVFA (R = 0.512, p = 0.030). Isobutyric acid showed positive correlations with protozoa, B. fibrisolvens, and F. succinogenes (p < 0.05). In summary, under the present experimental conditions, the addition of 0.5% L-Val to a low-nitrogen diet did not affect predicted CH₄ production, but improved other in vitro rumen fermentation parameters, including acetate, isobutyrate and MCP. Meanwhile, it favored the growth and proliferation of the fibrolytic bacteria (B. fibrisolvens and F. succinogenes). This provides a theoretical basis for the rational formulation of low-nitrogen diets for sheep. Full article

Underutilised, nutrient-dense legumes in their sprouted form provide promising substrates for developing functional fermented foods capable of influencing gut microbial activity and metabolite production. This study evaluated the effects of probiotic lactic acid bacteria-fermented beverages derived from sprouted Bambara groundnut (Vigna subterranea) and cowpea (Vigna unguiculata) on gut microbiota composition and short-chain fatty acid (SCFA) production using an in vitro colonic fermentation model. The beverages were fermented with either Bifidobacterium animalis BB-12 (BCBF24) or Lactiplantibacillus plantarum 75 (BCL7524). During colonic fermentation, at 0, 12, 24, and 38 h, faecal slurries were collected for SCFA analysis using gas chromatography–mass spectrometry (GC-MS) and deoxyribonucleic acid (DNA) sequencing (Oxford Nanopore Technologies). Microbial diversity decreased, indicating selective enrichment of taxa. BCL7524 induced a major shift, significantly (p < 0.05) enriching Bacillota and driving Megasphaera to ~42% dominance within 24 h. This reflected cross-feeding from L. plantarum to lactate-utilising Megasphaera spp. Spearman correlation linked Megasphaera to a broad SCFA profile, including isobutyric, isovaleric, valeric, and hexanoic acids, with a significant (p < 0.05) positive correlation observed for hexanoic acid. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated proteolysis and mapped hexanoic acid to fatty acid biosynthesis pathways, suggesting chain-elongation activity contributing to hexanoate formation. In line with this, BCL7524 produced significantly (p < 0.05) higher levels of hexanoate (3–14 mM) and valerate (10–15 mM), supporting chain-elongation activity within the community. In contrast, BCBF24 enriched Actinomycetota and Bifidobacterium, correlating with acetate production (18–23 mM). This study demonstrates that specific synbiotic beverages can modulate gut microbial ecology and metabolic output under in vitro conditions. Full article