Bioactive Potential Research Articles

Insights into the culturable fungal endophytes of Dioscorea bulbifera L. in terms of their diversity, antidiabetic and antioxidant activity

The research on endophytes is focused on understanding the complex microbiome compositions, their interactions with host plants and their bioactive potential. Our study provided an overview of the diversity, distribution and bioactivities of culturable endophytic fungi associated with fungal endophytes of Dioscorea bulbifera L. The phylogenetic analysis depicted the evolutionary relationship among taxa of endophytic fungal isolates. The isolated fungal endophytes belonged to twenty-five genera and thirty-eight species. Further, diversity indices demonstrated significant diversity of fungal endophytes in the different tissues: stem, leaf, tuber and bulbil. A higher consistency of endophytic fungal isolates in the bulbils was found using Shannon index. Furthermore, Simpson's index revealed that the leaf tissue harboured highly diverse fungal endophytes. Likewise, Margalef's index depicted high taxonomic richness in bulbils. The isolates such as Acrocalymma medicaginis, Curvularia lycoperscii, Talaromyces macrospora, Fusarium laceratum, Paecilomyces formosus and Microascus cirrosus isolated in this study have been reported as endophytes for the first time from any plant. In-vitro antioxidant and antidiabetic activities of the ethyl acetate extracts revealed that Nigrospora oryzae (Z2) effectively inhibited α-glucosidase activity with IC50 value of 0.6 µg/ml whereas IC50 value of the acarbose, the positive control was reported to be 0.040 µg/ml. The results of antioxidant activity demonstrated that H15A (Acrocalymma medicaginis) and BD5 (Phomopsis longicolla) could be used as potential sources of antioxidant agents. Moreover, H15A (Acrocalymma medicaginis) was reported to produce 687.1 ± 0.17 µg GAE/mg of TPC and 78.55 ± 0.29 µg QE/mg of TFC quantitatively. These findings suggest that the potential endophytes could be explored using systematic bio-guided investigations to further discover novel natural products. Our study provides an important resource for a deeper understanding of endophyte-plant interactions at molecular and genetic levels.

Discovery of Highly Bioactive Peptides through Hierarchical Structural Information and Molecular Dynamics Simulations.

Peptide drugs play an essential role in modern therapeutics, but the computational design of these molecules is hindered by several challenges. Traditional methods like molecular docking and molecular dynamics (MD) simulation, as well as recent deep learning approaches, often face limitations related to computational resource demands, complex binding affinity assessments, extensive data requirements, and poor model interpretability. Here, we introduce PepHiRe, an innovative methodology that utilizes the hierarchical structural information in peptide sequences and employs a novel strategy called Ladderpath, rooted in algorithmic information theory, to rapidly generate and enhance the efficiency and clarity of novel peptide design. We applied PepHiRe to develop BH3-like peptide inhibitors targeting myeloid cell leukemia-1, a protein associated with various cancers. By analyzing just eight known bioactive BH3 peptide sequences, PepHiRe effectively derived a hierarchy of subsequences used to create new BH3-like peptides. These peptides underwent screening through MD simulations, leading to the selection of five candidates for synthesis and subsequent in vitro testing. Experimental results demonstrated that these five peptides possess high inhibitory activity, with IC50 values ranging from 28.13 ± 7.93 to 167.42 ± 22.15 nM. Our study explores a white-box model driven technique and a structured screening pipeline for identifying and generating novel peptides with potential bioactivity.

Nanoencapsulation of creatine: Unlocking hydrophilic bioactive potential through double emulsification by solvent diffusion

AbstractNanotechnology has emerged as a pivotal tool in pharmaceutical research, driving the development of innovative techniques to enhance drug distribution within the body by finely tuning the biopharmaceutical properties of various drugs. Among these techniques, encapsulating biologically active substances in polymeric nanoparticles has garnered significant attention. However, achieving high encapsulation efficiency for hydrophilic molecules remains a formidable challenge, despite their growing importance in treating diverse diseases such as cancer, where nucleic acids, peptides, proteins, and smaller hydrophilic molecules are key players. While some studies have reported improvements in encapsulating hydrophilic drugs, notable examples, like the solid/oil/water ion‐pairing method and the use of calcium phosphate, have shown promise in enhancing encapsulation efficiency for different drugs. Nonetheless, the challenge of preparing aqueous core nanoparticles capable of encapsulating a high percentage of water‐soluble actives persists. Creatine monohydrate, a bioactive compound widely consumed by athletes for its role in increasing muscular phosphocreatine stores, presents an intriguing case. Despite its insolubility in organic media, creatine exhibits limited aqueous solubility, rendering it a suitable candidate for encapsulation in polymeric nanoparticles to improve its aqueous solubility and gastrointestinal absorption. In this study, we aim to produce polymeric nanoparticles containing creatine monohydrate utilizing the double emulsification (W/O/W) method. This promising and efficient approach holds the potential to significantly enhance the aqueous solubility and gastrointestinal absorption of creatine, thereby broadening its clinical application spectrum. Leveraging nanotechnology in this context offers an innovative and potentially impactful strategy to augment the therapeutic efficacy of creatine.

Chemical Constituents from the Whole Plants of Limonium sinense and Their in vitro Anti-inflammatory Activity.

The Yellow River Delta possesses lots of characteristic medicinal plants due to its high salinity and high alkaline environment and Limonium sinense is an iconic plant. However, there are very few studies on L. sinense and its chemical constituents have not been investigated in recent ten years. In the present study, the chemical constituents and bioactivities of L. sinense were fully studied for the first time. UPLC-MS/MS method combined with database comparison identified 109 compounds mainly including flavonoids, alkaloids and polyphenols. In addition, the potential bioactivities of L. sinense were considerated as anti-inflammatory, anti-oxidative, anti-tumor, hepatoprotective and hpyerglycemic activities based on these identified compounds and their related literature. Furthermore, four derivatives of 12-oxo-phytodienoic acid and butenolide including two new ones (1 and 2) were isolated from the whole plants of L. sinense. Their structures, including the absolute configurations, were determined by the analysis of comprehensive spectroscopic data. All isolates were evaluated for their anti-inflammatory activity. Compound 1 exhibited moderate anti-inflammatory activity with IC50 value of 37.5±1.2 μM on NO production level.

Bushy-Tailed Multicationic Quaternary Phosphonium Compounds: Potent Amphiphilic Disinfectants with Promising Therapeutic Indices.

Quaternary ammonium compounds (QACs) have been essential for protecting human health for almost a century, functioning as surface disinfectants and sanitizers. With bacterial resistance increasing against commercially available QACs, the development of novel antimicrobials with divergent architectures is essential for effective infection prevention and control. Toward this end, our group has expanded beyond traditional ammonium scaffolds and explored the development of quaternary phosphonium compounds (QPCs). Herein, we report the synthesis and biological investigation of a series of 20 novel multicationic QPCs, bearing multiple short alkyl or aryl chains, also referred to as "bushy-tailed" multiQPCs; these structures were hypothesized to have strong bioactivity while displaying low mammalian toxicity. Select bushy-tailed QPC derivatives with trishexylphosphonium groups displayed single-digit or sub-micromolar activity against all seven bacteria tested, and MIC values of 2- to 8-fold better than their bushy-tailed QAC counterparts. Importantly, therapeutic indices of these bushy-tailed QPCs were favorable in many cases, and were ≥ 4 against the entire bacterial panel for pX-P6*,P6* and 1,8-P6*,P6*, superior to more traditional architectures. This work highlights the promise of a novel set of multicationic phosphonium compounds as novel disinfectants with potent bioactivities and low toxicity.

Green synthesis, molecular docking and in vitro biological evaluation of novel hydrazones, pyrazoles, 1,2,4-triazoles and 1,3,4-oxadiazoles

Ultrasonic waves were used for synthesis of novel hydrazones, bishydrazones, pyrazoles, 1,2,4-triazole and 1,3,4-oxadiazole. The structural formulae of the synthesized compounds were elucidated in terms of elemental and spectroscopic analyses. All synthesized compounds were estimated by testing total antioxidant capacity, iron‐reducing power, the scavenging activity against ABTS and DPPH radicals in addition to testing anti‐diabetic, anti‐Alzheimer and anti‐arthritic activities. All compounds displayed good to potent bioactivity. Compounds 6, 10 exhibit the highest antioxidant and free radicals scavenging activities. Furthermore, compounds 6, 10 demonstrate the strongest inhibition of α‐amylase and α-glucosidase. Compound 12 exhibit strongest acetylcholinesterase inhibition among prepared compounds. However, compounds 18a,b, 19 show a significantly higher inhibition percentage for protein denaturation and proteinase. The most bioactive prepared compounds 6, 10 and 12 were investigated toward docking methodology against appropriate protein.

Bioactive Potential of Probiotic Bacteria-A Comprehensive Review

Bioactive Potential of Probiotic Bacteria-A Comprehensive Review

Effect of Fermentation by Probiotic Bacteria on the Bioaccessibility of Bioactive Compounds from the Fruit of the Juçara Palm (Euterpe edulis Martius)

The underexplored fruit from the juçara palm tree (Euterpe edulis Martius) has bioactive compounds with antioxidant activities, such as phenolic acids and anthocyanins. This fruit’s pulp presents itself as an appropriate fermentation medium for probiotic bacteria growth. Therefore, this study was conducted to evaluate the effects of fermentation by Limosilactobacillus reuteri LR92 (JLR) and Bifidobacterium animalis ssp. lactis BB-12 (JBB) on the bioactive compound contents of the juçara pulp, before and after a gastrointestinal simulation. The pulp of the juçara fruit showed probiotic counts of 8.70 ± 0.07 log UFC/mL for JLR and 8.44 ± 0.09 log UFC/mL for JBB, after 24 h of fermentation. Fermentation with the strains used modified the proportions of fatty acids (fatty acids esters were quantified using a gas chromatography equipment) and fibers when compared to the non-fermented pulp. The antioxidant capacity determined by DPPH, FRAP and ABTS showed significant reduction after the gastrointestinal simulation for samples. Phenolic compound analysis by UPLC-MS/MS showed, after fermentation, a greater amount of ferulic, protocatechuic and catechin acids in the samples. These results show changes in the bioactive compounds due to the fermentation of the juçara pulp by probiotics. However, these compounds showed bioactive potential and were bioaccessible after the gastrointestinal simulation, with the pulp being a potential means for bacteria growth, which may bring health benefits.

Isolation, Identification and Pigment Analysis of Novel Cyanobacterial Strains from Thermal Springs

Cyanobacterial pigments have attracted considerable attention in industry due to their bioactive potential and natural origin. In the present study, the growth dynamics and pigment composition, in terms of chlorophyll a, total carotenoids and phycobiliprotein content, of four cyanobacterial strains isolated from thermal springs, namely Oscillatoria subbrevis CZS 2201, Phormidium ambiguum CZS 2205, Nostoc calcicola TSZ 2203, and Synechococcus sp. CZS 2204, were investigated. The analysis revealed that the maximum quantity of chlorophyll a and total carotenoids was observed in Oscillatoria subbrevis CZS 2201 (26.49 and 3.44 µg mL−1), followed by Phormidium ambiguum CZS 2205 (18.64 and 2.32 µg mL−1), whereas a minimum amount was detected in Synechococcus sp. CZS 2204 (12.13 and 1.24 µg mL−1), respectively. In addition, Oscillatoria subbrevis CZS 2201 showed higher quantity of phycobiliproteins, especially C-phycocyanin (45.81 mg g−1), C-phycoerythrin (64.17 mg g−1) and C-allophycocyanin (27.45 mg g−1). Moreover, carotenoid derivatives of Oscillatoria subbrevis CZS 2201 were also identified, among which β-carotene was the dominant form (1.94 µg mL−1), while the accumulation of zeaxanthin and myxoxanthophyll was relatively high (0.53 and 0.41 µg mL−1, respectively) compared with echinenone and cryptoxanthin (0.34 and 0.23 µg mL−1, respectively). The study revealed that Oscillatoria subbrevis CZS 2201 was a potent producer of secondary carotenoids, including myxoxanthophyll.

Combining culture optimization and synthetic biology to improve production and detection of secondary metabolites in Myxococcus xanthus: application to myxoprincomide.

Microbial secondary metabolites play crucial ecological roles in governing species interactions and contributing to their defense strategies. Their unique structures and potent bioactivities have been key in discovering antibiotics and anticancer drugs. Genome sequencing has undoubtedly revealed that myxobacteria constitute a huge reservoir of secondary metabolites as the well-known producers, actinomycetes. However, because most secondary metabolites are not produced in the laboratory context, the natural products from myxobacteria characterized to date represent only the tip of the iceberg. By combining the engineering of a dedicated Myxococcus xanthus DZ2 chassis strain with a two-step growth medium protocol, we provide a new approach called two-step Protocol for Resource Integration and Maximization-Biomolecules Overproduction and Optimal Screening Therapeutics (2PRIM-BOOST) for the production of non-ribosomal peptides synthetases (NRPS)/polyketides synthases (PKS) secondary metabolites from myxobacteria. We further show that the 2PRIM-BOOST strategy will facilitate the screening of secondary metabolites for biological activities of medical interest. As proof of concept, using a constitutive strong promoter, the myxoprincomide from M. xanthus DZ2 has been efficiently produced and its biosynthesis has been enhanced using the 2PRIM-BOOST approach, allowing the identification of new features of myxoprincomide. This strategy should allow the chances to produce and discover new NRPS, PKS, and mixed NRPS/PKS hybrid natural metabolites that are currently considered as cryptic and are the most represented in myxobacteria.IMPORTANCEMicrobial secondary metabolites are important in species interactions and are also a prolific source of drugs. Myxobacteria are ubiquitous soil-dwelling bacteria constituting a huge reservoir of secondary metabolites. However, because most of these molecules are not produced in the laboratory context, one can estimate that only one-tenth have been characterized to date. Here, we developed a new strategy called two-step Protocol for Resource Integration and Maximization-Biomolecules Overproduction and Optimal Screening Therapeutics (2PRIM-BOOST) that combines the engineering of a dedicated Myxococcus xanthus chassis strain together with growth medium optimization. By combining these strategies with the insertion of a constitutive promoter upstream the biosynthetic gene cluster (BGC), the production of myxoprincomide, a characterized low-produced secondary metabolite, was successfully and significantly increased. The 2PRIM-BOOST enriches the toolbox used to produce previously cryptic metabolites, unveil their ecological role, and provide new molecules of medical interest.

Stereodivergent Synthesis of Chiral Hydrobenzofuranpyrrolidines by Catalytic Asymmetric Dearomative Cyclization and Controlled Epimerization

A rapid and straightforward way to access stereoisomeric sets of products bearing multiple stereogenic centers is still a significant challenge in asymmetric catalysis. We present herein our experimental studies on the stereodivergent synthesis of chiral hydrobenzofuran‐fused pyrrolidines with three stereogenic centers via organocatalytic asymmetric dearomative cyclization and epimerization process. Chiral bifunctional thiourea catalyst could successfully promote the enantioselective dearomatization cyclization of 2‐nitrobenzofurans with o‐hydroxy aromatic aldimines, which enabled the synthesis of (3S,3aR,8bR)‐hydrobenzofuran[3.2]pyrrolidines in 79‐92% yields with >20:1 stereoselectivities and 93‐99% enantio‐ selectivities. While catalytic amount of DBU could induce the direct intramolecular epimerization of (3S,3aR,8bR)‐hydrobenzofuran[3.2] pyrrolidines to its diastereomers (3R,3aR,8bR)‐hydrobenzofuran[3.2] pyrrolidines in 72‐87% yields without loss of stereoselectivities. The mechanistic pathways of the epimerization process were investigated by a series of control experiments study. This work provides an alternative and forward solution for the stereodivergent preparation of functionalized pyrrolidines with potential bioactivities.

The genus Haplopappus: botany, phytochemistry, traditional uses, and pharmacological properties

BackgroundThe genus Haplopappus Cass. [Asteraceae] comprises a large number of species distributed mainly in Chile and with various traditional medicinal uses.PurposeThe present review addresses the botany, traditional uses, chemistry, biological and pharmacological activities of the genus, aiming to further potentiate the associated research and applications.Study design and MethodsLiterature data on the chemistry and bioactivity of the genus Haplopappus were mainly retrieved from digital databases such as SciFinder®, PubMed®, and Google Scholar®, as well as from the scientific journal publishers’ platforms linked with these databases.Results and discussionAlthough the majority of the botanical taxa of the genus Haplopappus has been understudied, available information is promising regarding its phytochemistry and bioactivity. A total of more than 400 compounds are present in different Haplopappus species, mostly terpenoids and phenolic compounds. Scientific literature supports various health promoting effects of Haplopappus extracts and isolated compounds, principally their effect against human pathogenic bacteria and their high antioxidant capacity. The existing limitations highlighted hereby are mainly associated to the lack of modern investigation regarding a wider number of Haplopappus species and chemical compounds, as well as to the absence of in vivo bioactivity results and clinical trials.ConclusionScientific literature supports the ethnopharmacological, phytochemical and bioactive potential of the genus Haplopappus, however the aforementioned limitations need to be addressed in order to further promote and broaden both scientific research and future applications and uses.

Hot node limitations and impact of taxonomic resolution on phylogenetic divergence patterns: A case study on Ecuadorian ethnomedicinal flora

SummarySocietal Impact StatementNovel phylogenetic approaches have emerged in recent years to study traditional medicinal plants, aiming to identify potential sources of new drugs. This line of research holds considerable promise yet remains in its early stages. Here, we examine the prevalent methods employed in the field, revealing the impact of methodological choices that have often been made arbitrarily. We also highlight a widespread misconception regarding ‘hot node analysis’, a tool gaining popularity for identifying plants with high bioactive potential. Our findings should advance future research aimed at guiding the selection of promising candidates for bioprospection.Summary Advances in phylogenetics offer a ground‐breaking approach to analysing ethnobotanical data. This line of research typically involves calculating the degree of phylogenetic clustering (i.e. phylogenetic divergence) of a set of medicinal plants and identifying clades with a significant overabundance of these plants, known as hot nodes, which are purportedly responsible for the clustering patterns. However, despite showing great promise, the adequacy of this procedure remains to be tested, and the results have so far been inconclusive and, at times, contradictory. Here, we examine two key elements through a case study of Ecuadorian medicinal flora: the impact of taxonomic resolution (species‐ and genus‐level data) on phylogenetic divergence metrics (Mean Pairwise Distance; MPD, and Mean Nearest Taxon Distance; MNTD), and the efficacy of the hot node analysis in identifying clades that significantly influence these metrics. To identify clades with a significant impact on MPD and MNTD, we implemented a jackknifing procedure and compared outcomes with the hot nodes. Phylogenetic divergence was strongly dependent on the taxonomic resolution, with clustering mainly revealed by MPD at the genus level. Further, the hot nodes incompletely matched those with significant influence on MPD and MNTD according to the jackknifing analysis. We highlight the impact of taxonomic resolution on commonly used phylogenetic divergence metrics and the limitations of hot nodes in identifying influential clades, stressing the potential of alternative jackknifing techniques. We recommend the use of the most resolved tree possible and combining jackknifing with hot node analyses as complementary sources to pinpoint clades for bioprospection.

Revealing the Potential of Star Anise Essential Oil: Comparative Analysis and Optimization of Innovative Extraction Methods for Enhanced Yield, Aroma Characteristics, Chemical Composition, and Biological Activities

ABSTRACTStar anise has been used for a long time in improving human health and curing diseases, owing to its unlimited components with complex chemical structures and a wide range of bioactivities. This study is aimed to investigate the influence of extraction methods (steam distillation, ethanol Soxhlet extraction, supercritical carbon dioxide extraction, subcritical n‐butane extraction) on the yield, aroma properties, chemical composition, and bioactivity of star anise essential oils. Electronic nose detection revealed the essential oils from subcritical extraction exhibited the most intense aroma, while the essential oils from ethanol Soxhlet extraction had a more complex aroma profile. Fourier‐transform infrared analysis showed the presence of benzene rings, carbonyl groups, C=C, and aromatic ether bonds in the essential oils extracted through different methods. The major components were heterocyclic olefins, heterocyclic oxygenates, and aromatic oxygenates, as well as certain amounts of flavonoids and polyphenols. Correlation analysis revealed the relative contents of volatile trans‐anethole, wormwood, d‐limonene, cineole, and trans‐α‐citronelene were strongly associated with the antibacterial activity of the essential oils. Similarly, the contents of volatile components (d‐limonene, cineole) and non‐volatile components (total flavonoids and total polyphenols) were strongly correlated with the DPPH scavenging activity of the essential oils. These results confirm the effectiveness of the ethanol Soxhlet extraction method in retaining the bioactivity of the essential oils. Finally, with a Box–Behnken central composite design of response surface and single‐factor experiments, the optimal extraction conditions for the ethanol Soxhlet method were determined: ultrasonic frequency of 80 kHz, crushing particle size of 60 meshes, liquid–material ratio of 8:1 (mL:g), and ultrasonic time of 35 min. Under these conditions, the essential oil yield was 25.51% ± 0.21%. Overall, these findings highlight the significance of extraction methods in obtaining high‐quality star anise essential oils with desirable aroma properties and potent bioactivities.

Effect of the extraction method on the lipid composition of purslane (Portulaca oleracea L.) seed oil

Purslane (Portulaca oleracea L.) is a widespread weed plant used since ancient times as remedy and as food. Its seed oil possesses significant bioactive potential due to the high content of important phytonutrients, mainly essential fatty acids and phytosterols. Although the total fatty acid composition of oil is well documented, till now there is no data published about individual lipid classes. The information on sterols is scarce as well. Therefore, the aim of this work was to characterize in details for the first time the individual lipid classes and their fatty acid composition, sterols, as well as the oxidative stability of the oil, depending on the extraction method by either hexane, chloroform–methanol mixtures or super-critical CO2. The results revealed no significant effect of extraction method on the individual lipid classes (monoacylglycerols, diacylglycerols, free fatty acids, free sterols, triacylglycerols, sterol esters, wax esters and hydrocarbons), including the fatty acid composition of saponifiables, and quite weak effect on the oxidative stability of the oil regarding super-critical CO2 vs. organic solvents. Detailed analyses of lipid composition confirmed the potential of purslane seed oil as a cheap and highly valuable source of phytonutrients as essential fatty acids and phytosterols, for application in food, pharmaceutical and cosmetic industries.

Unveiling the Potential Bioactive Peptides Derived From Goat Casein Hydrolysates Based on In Silico Analyses

ABSTRACTBioactive peptides (BAPs) have attracted considerable interest in scientific research due to their heterogeneity in sequence and structure, which underpins various biological functionalities. In this context, goat casein, an abundant by‐product of the dairy industry, emerges as a valuable source of BAPs. The present study undertook a meticulous evaluation of the bioactive potentials of goat casein‐derived peptides through an integrated approach combining computational simulations, high‐throughput screening, and molecular docking techniques. The initial phase involved the enzymatic digestion of goat milk casein using trypsin, followed by the identification of peptides via liquid chromatography–tandem mass spectrometry (LC–MS/MS), uncovering a total of 597 peptides. Subsequent prioritization using the PeptideRanker algorithm identified 70 peptides exhibiting potential bioactivity, as denoted by scores above 0.8. Advanced screening employing the BIOPEP database and the AutoDock and CAMPR4 tools facilitated the elucidation of 16 antioxidant, 59 hypotensive, 63 hypoglycemic, 70 hypolipidemic, and 25 antimicrobial peptides. Molecular docking studies further elucidated the spontaneous nature of the interactions between the peptides and their respective receptors, predominantly mediated by hydrogen bonding and hydrophobic interactions. Four peptides specifying all activities simultaneously were synthesized, and their activities were verified by in vitro experiments. These results not only highlight an effective strategy for the high‐throughput screening of goat casein‐derived peptides but also underscore the potential of utilizing casein as a viable source of functional food ingredients. This study thereby contributes significantly to the expanding field of functional food research, suggesting a sustainable approach to explore the potential of dairy by‐products.

Exploring the bioactivity potential of indigenous Ganoderma species from Northern Serbia: insights into antioxidant and anti-acetylcholinesterase properties.

This study explores the bioactivity potential of four indigenous Ganoderma species from northern Serbia (G. applanatum, G. lucidum, G. pfeifferi, and G. resinaceum), comparing antioxidant and anti-acetylcholinesterase (anti-AChE) properties. The mycochemical profile, including ethanol (EtOH), water (H2O), and chloroform (CHCl3) extracts, was analysed, revealing significant phenolic compounds, such as 2,5-dihydroxybenzoic acid. Highest total phenolic content (TP) was in EtOH extracts (205 - 380 mg GAE/g d.w.), and total carbohydrate content peaked in G. pfeifferi CHCl3 extracts (1895 ± 0.29 mg GluE/g d.w., 439 ± 0.42 mg FruE/g d.w.). DPPH and FRAP activities were prominent in G. applanatum H2O extracts, while CHCl3 extracts excelled in the NO assay, and EtOH extracts were superior in the ABTS assay. The most effective AChE inhibitors were polar extracts from G. pfeifferi. These findings emphasise the potential therapeutic use of Ganoderma species, particularly G. pfeifferi, in combating oxidative stress and acetylcholine deficiencies.

The Green Extraction of Blueberry By-Products: An Evaluation of the Bioactive Potential of the Anthocyanin/Polyphenol Fraction.

In the context of a circular economy, this study explores the valorization of blueberry pomace (BP) as a source of bioactive compounds using sustainable extraction methods. Microwave-assisted extraction (MAE) and microwave-assisted subcritical water extraction (MASWE) were employed to obtain two distinct fractions: MAE 1° and MASWE 2°. The first extract, MAE 1°, obtained at 80 °C, had a high total anthocyanin content (21.96 mgCya-3-glu/gextract), making it suitable as a natural pigment. Additionally, MAE 1° exhibited significant enzyme inhibition, particularly against α-amylase and β-glucosidase, suggesting potential anti-diabetic and anti-viral applications. The second extract, MASWE 2°, obtained at 150 °C, contained a higher total phenolic content (211.73 mgGAE/gextract) and demonstrated stronger antioxidant activity. MASWE 2° showed greater inhibition of acetylcholinesterase and tyrosinase, indicating its potential for use in Alzheimer's treatment, skincare, or as a food preservative. MASWE 2° exhibited cytotoxicity against HeLa cells and effectively mitigated H2O2-induced oxidative stress in HaCat cells, with MAE 1° showing similar but less pronounced effects. A tested formulation combining MAE 1° and MASWE 2° extracts in a 3:2 ratio effectively enhanced anthocyanin stability, demonstrating its potential as a heat-stable pigment. The extract characteristics were compared with a conventional method (MeOH-HCl in reflux condition), and the protocol's sustainability was assessed using several green metric tools, which provided insights into its environmental impact and efficiency.

Association Between Dairy Production System and Milk Functionality Based on Analysis of miRNAs in Exosomes from Milk.

Dairy farming practices significantly affect the nutritional and functional properties of milk. This study compares miRNAs in milk exosomes from extensive and intensive dairy systems and explores their potential implications for human consumers. Extensive systems are believed to produce milk of higher quality with better animal welfare compared to intensive systems. Milk samples from eight extensive and nine intensive dairy farms were analysed. Milk-derived extracellular vesicles were isolated through sequential ultracentrifugation and characterised through Dynamic Light Scattering and Nanosight to determine the size and the concentration of the extracellular vesicles, in addition to immunoblotting to ensure the presence of exosome-specific proteins in their membrane. miRNA levels were quantified using RT-qPCR, and metabolic pathways associated with miRNAs showing significant differences between farm groups were analysed. EVs from extensive farms had higher concentrations. Notably, bta-miR-451 levels were significantly higher in milk from extensive farms (p = 0.021). Like human miRNA hsa-miR-451, it is linked to pathways related to Parkinson's disease and cancer. Our research suggests that milk production in extensive systems not only provides socioeconomic and environmental benefits but may also have positive effects on human health. Further research is warranted to explore the bioactive potential of these miRNAs and their implications for human health.

Limosilactobacillus reuteri DSM 17938 Produce Bioactive Components during Formulation in Sucrose.

Improved efficacy of probiotics can be achieved by using different strategies, including the optimization of production parameters. The impact of fermentation parameters on bacterial physiology is a frequently investigated topic, but what happens during the formulation, i.e., the step where the lyoprotectants are added prior to freeze-drying, is less studied. In addition to this, the focus of process optimization has often been yield and stability, while effects on bioactivity have received less attention. In this work, we investigated different metabolic activities of the probiotic strain Limosilactobacillus reuteri DSM 17938 during formulation with the freeze-drying protectant sucrose. We discovered that the strain consumed large quantities of the added sucrose and produced an exopolysaccharide (EPS). Using NMR, we discovered that the produced EPS was a glucan with α-1,4 and α-1,6 glycosidic bonds, but also that other metabolites were produced. The conversion of the lyoprotectant is hereafter designated lyoconversion. By also analyzing the samples with GCMS, additional potential bioactive compounds could be detected. Among these were tryptamine, a ligand for the aryl hydrocarbon receptor, and glycerol, a precursor for the antimicrobial compound reuterin (3-hydroxypropionaldehyde). To exemplify the bioactivity potential of lyoconversion, lyoconverted samples as well as purified EPS were tested in a model for immunomodulation. Both lyoconverted samples and purified EPS induced higher expression levels of IL-10 (2 times) and IL-6 (4-6 times) in peripheral blood mononuclear cells than non-converted control samples. We further found that the initial cultivation of DSM 17938 with sucrose as a sugar substrate, instead of glucose, improved the ability to convert sucrose in the lyoprotectant into EPS and other metabolites. Lyoconversion did not affect the viability of the bacteria but was detrimental to freeze-drying survival, an issue that needs to be addressed in the future. In conclusion, we show that the metabolic activities of the bacteria during the formulation step can be used as a tool to alter the activity of the bacteria and thereby potentially improve probiotic efficacy.