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Parasites & Vectors Jul 2023Helminth infections are an important public health problem in humans and have an even greater impact on domestic animal and livestock welfare. Current readouts for...
BACKGROUND
Helminth infections are an important public health problem in humans and have an even greater impact on domestic animal and livestock welfare. Current readouts for anthelmintic drug screening assays are stage development, migration, or motility that can be subjective, laborious, and low in throughput. The aim of this study was to apply and optimize a fluorometric technique using resazurin for evaluating changes in the metabolic activity of Ascaris suum third-stage larvae (L3), a parasite of high economic relevance in swine.
METHODS
Ascaris suum L3 were mechanically hatched from 6- to 8-week embryonated and sucrose-gradient-enriched eggs. Resazurin dye and A. suum L3 were titrated in 96-well microtiter plates, and resazurin reduction activity was assessed by fluorometry after 24 h of incubation. Fluorescence microscopy was used to localize the resazurin reduction site within the larvae. Finally, we exposed A. suum L3 to various stress conditions including heat, methanol, and anthelmintics, and investigated their impact on larval metabolism through resazurin reduction activity.
RESULTS
We show that the non-fluorescent dye resazurin is reduced inside vital A. suum L3 to fluorescent resorufin and released into the culture media. Optimal assay parameters are 100-1000 L3 per well, a resazurin concentration of 7.5 µg/ml, and incubation at 37 °C/5% CO for 24 h. An intact L2 sheath around the L3 of A. suum completely prevents the uptake of resazurin, while in unsheathed L3, the most intense fluorescence signal is observed along the larval midgut. L3 exposed to methanol or heat show a gradually decreased resazurin reduction activity. In addition, 24 h exposure to ivermectin at 0.625 µM, mebendazole at 5 µM, and thiabendazole from 10 to 100 µM significantly decreased larval metabolic activity by 55%, 73%, and 70% to 89%, respectively.
CONCLUSIONS
Together, our results show that both metabolic stressors and anthelmintic drugs significantly and reproducibly reduce the resazurin reduction activity of A. suum L3, making the proposed assay a sensitive and easy-to-use method to evaluate metabolic activity of A. suum L3 in vitro.
Topics: Humans; Animals; Swine; Ascaris suum; Methanol; Anthelmintics; Xanthenes; Ascariasis; Larva
PubMed: 37468906
DOI: 10.1186/s13071-023-05871-5 -
Molecules (Basel, Switzerland) Dec 2023is a widely distributed crust fungus reported from all continents except Antarctica, although considered a rare species in several European countries. Profound...
is a widely distributed crust fungus reported from all continents except Antarctica, although considered a rare species in several European countries. Profound mycochemical analysis of the methanol extract of resulted in the isolation of seven compounds (-). Among them, (3β,22E)-3-methoxy-ergosta-4,6,8,22-tetraene () is a new natural product, while the NMR assignment of its already known epimer () has been revised. In addition to a benzohydrofuran derivative fomannoxin (), four ergostane-type triterpenes - were identified. The structure elucidation of the isolated metabolites was performed by one- and two-dimensional NMR and MS analysis. Compounds - as well as the chloroform, -hexane, and methanol extracts of were evaluated for their tyrosinase, acetylcholinesterase, and butyrylcholinesterase inhibitory properties. Among the examined compounds, only fomannoxin () displayed the antityrosinase property with 51% of inhibition, and the fungal steroids proved to be inactive. Regarding the potential acetylcholinesterase (AChE) inhibitory activity of the fungal extracts and metabolites, it was demonstrated that the chloroform extract and compounds - exerted noteworthy inhibitory activity, with 83.86 and 32.99%, respectively. The butyrylcholinesterase (BChE) inhibitory assay revealed that methanol and chloroform extracts, as well as compounds and exerted notable activity, while the rest of the compounds proved to be only weak enzyme inhibitors. Our study represents the first report on the chemical profile of basidiome of the wild-growing , offering a thorough study on the isolation and structure determination of the most characteristic biologically active constituents of this species.
Topics: Cholinesterase Inhibitors; Acetylcholinesterase; Butyrylcholinesterase; Chloroform; Methanol; Basidiomycota; Plant Extracts
PubMed: 38202796
DOI: 10.3390/molecules29010213 -
Veterinary Medicine and Science Jul 2023Plants are widely used in traditional medicine because they contain a high concentration of antimicrobial agents, serving as the foundation for medicines. The aim of...
INTRODUCTION
Plants are widely used in traditional medicine because they contain a high concentration of antimicrobial agents, serving as the foundation for medicines. The aim of this study was preliminary identification of phytochemicals and assesses the antimicrobial activity of extracts of Ferula communis root bark.
METHODS
Plant was collected, and standard qualitative procedures were conducted. The plant samples were extracted with 99.9% methanol and 80% ethanol. To identify phytochemicals found in plants, a preliminary phytochemical analysis was performed. Agar diffusion tests, minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were performed to evaluate antibacterial activity.
RESULT
The preliminary phytochemical analysis of the ethanol and methanol extract revealed positive results for flavonoids, coumarins and tannins. Terpenoids and anthraquinones were detected only in the methanol extract. The extract of Ferula communis showed an antibacterial effect on both gram-negative and gram-positive bacteria in a concentration-dependent manner. The average zone of inhibition for gram-positive bacteria was 11 mm, whereas for gram-negative bacteria, it was 9 mm. The MIC and MBC values also varied with the type of bacteria. In all bacterial species tested, the mean MBC value was similar to the MIC.
CONCLUSION
Different phytochemicals were detected in extracts of the root bark of F. communis and extracts showed antibacterial effects in a concentration-dependent manner. Therefore, further purification and evaluation of the extracts and antioxidant activity of the plant should be investigated.
Topics: Animals; Plant Extracts; Ferula; Apiaceae; Methanol; Plant Bark; Anti-Bacterial Agents; Phytochemicals; Bacteria; Gram-Positive Bacteria; Ethanol
PubMed: 37392454
DOI: 10.1002/vms3.1170 -
Chemical Reviews Feb 2024Methane is a potent greenhouse gas that contributes significantly to climate change and is primarily regulated in Nature by methanotrophic bacteria, which consume... (Review)
Review
Methane is a potent greenhouse gas that contributes significantly to climate change and is primarily regulated in Nature by methanotrophic bacteria, which consume methane gas as their source of energy and carbon, first by oxidizing it to methanol. The direct oxidation of methane to methanol is a chemically difficult transformation, accomplished in methanotrophs by complex methane monooxygenase (MMO) enzyme systems. These enzymes use iron or copper metallocofactors and have been the subject of detailed investigation. While the structure, function, and active site architecture of the copper-dependent particulate methane monooxygenase (pMMO) have been investigated extensively, its putative quaternary interactions, regulation, requisite cofactors, and mechanism remain enigmatic. The iron-dependent soluble methane monooxygenase (sMMO) has been characterized biochemically, structurally, spectroscopically, and, for the most part, mechanistically. Here, we review the history of MMO research, focusing on recent developments and providing an outlook for future directions of the field. Engineered biological catalysis systems and bioinspired synthetic catalysts may continue to emerge along with a deeper understanding of the molecular mechanisms of biological methane oxidation. Harnessing the power of these enzymes will necessitate combined efforts in biochemistry, structural biology, inorganic chemistry, microbiology, computational biology, and engineering.
Topics: Copper; Methane; Iron; Methanol; Oxygenases; Oxidation-Reduction; Mixed Function Oxygenases
PubMed: 38305159
DOI: 10.1021/acs.chemrev.3c00727 -
Nanomaterials (Basel, Switzerland) Nov 2023CuZn-based catalysts are widely used in CO hydrogenation, which may effectively convert CO to methanol and alleviate CO emission issues. The precise design of a model...
CuZn-based catalysts are widely used in CO hydrogenation, which may effectively convert CO to methanol and alleviate CO emission issues. The precise design of a model catalyst with a clear atomic structure is crucial in studying the relationship between structure and catalytic activity. In this work, a one-pot strategy was used to synthesize CuZn@ZSM-5 catalysts with approximately two Cu atoms and one Zn atom per unit cell. Atomic Cu and Zn species are confirmed to be located in the [5.6.10] and [6.10] tilings, respectively, by using magic-angle spinning nuclear magnetic resonance spectroscopy (MAS NMR), synchrotron X-ray powder diffraction (SXRD) and high-signal-to-noise-ratio annular dark field scanning transmission electron microscopy (High SNR ADF-STEM). Catalytic hydrogenation of CO to methanol was used as a model reaction to investigate the activity of the catalyst with confined active species. Compared to the Cu@ZSM-5, Zn@ZSM-5 and their mixture, the CuZn@ZSM-5 catalyst with a close Cu-Zn distance of 4.5 Å achieves a comparable methanol space-time yield (STY) of 92.0 mg·g·h at 533 K and 4 MPa with high stability. This method is able to confine one to three metal atoms in the zeolite channel and avoid migration and agglomeration of the atoms during the reaction, which maintains the stability of the catalyst and provides an efficient way for adjustment of the type and number of metal atoms along with the distances between them in zeolites.
PubMed: 38063749
DOI: 10.3390/nano13233053 -
Synthetic and Systems Biotechnology Sep 2023Methanol is a promising one-carbon feedstock for biomanufacturing, which can be sustainably produced from carbon dioxide and natural gas. However, the efficiency of...
Methanol is a promising one-carbon feedstock for biomanufacturing, which can be sustainably produced from carbon dioxide and natural gas. However, the efficiency of methanol bioconversion is limited by the poor catalytic properties of nicotinamide adenine dinucleotide (NAD)-dependent methanol dehydrogenase (Mdh) that oxidizes methanol to formaldehyde. Herein, the neutrophilic and mesophilic NAD-dependent Mdh from DSM 2334 (Mdh) was subjected to directed evolution for enhancing the catalytic activity. The combination of formaldehyde biosensor and Nash assay allowed high-throughput and accurate measurement of formaldehyde and facilitated efficient selection of desired variants. Mdh variants with up to 6.5-fold higher / value for methanol were screened from random mutation libraries. The T153 residue that is spatially proximal to the substrate binding pocket has significant influence on enzyme activity. The beneficial T153P mutation changes the interaction network of this residue and breaks the α-helix important for substrate binding into two short α-helices. Reconstructing the interaction network of T153 with surrounding residues may represent a promising strategy to further improve Mdh, and this study provides an efficient strategy for directed evolution of Mdh.
PubMed: 37342805
DOI: 10.1016/j.synbio.2023.05.004 -
Communications Chemistry Dec 2023The direct oxidation of methane to methanol has been spotlighted research for decades, but has never been commercialized. This study introduces cost-effective process...
The direct oxidation of methane to methanol has been spotlighted research for decades, but has never been commercialized. This study introduces cost-effective process for co-producing methanol and sulfuric acid through a direct oxidation of methane. In the initial phase, methane oxidation forms methyl bisulfate (CHOSOH), then transformed into methyl trifluoroacetate (CFCOCH) via esterification, and hydrolyzed into methanol. This approach eliminates the need for energy-intensive separation of methyl bisulfate from sulfuric acid by replacing the former with methyl trifluoroacetate. Through the superstructure optimization, our sequential process reduces the levelized cost of methanol to nearly two-fold reduction from the current market price. Importantly, this process demonstrates adaptability to smaller gas fields, assuring its economical operation across a broad range of gas fields. The broader application of this process could substantially mitigate global warming by utilizing methane, leading to a significantly more sustainable and economically beneficial methanol industry.
PubMed: 38123721
DOI: 10.1038/s42004-023-01080-4 -
Frontiers in Chemistry 2023In this study, aqueous, ethanol, methanol, and hexane extracts from Iraqi Kurdistan Region Daphne mucronata were prepared due to the numerous applications and...
Methanol extract of Iraqi Kurdistan Region as a potent source of antioxidant, antimicrobial, and anticancer agents for the synthesis of novel and bioactive polyvinylpyrrolidone nanofibers.
In this study, aqueous, ethanol, methanol, and hexane extracts from Iraqi Kurdistan Region Daphne mucronata were prepared due to the numerous applications and development of nanofibers in biological and medical fields, including food packaging, enzyme stabilization, and wound dressing. In the initial evaluation of the extracts, the antioxidant properties against DPPH, antimicrobial properties against 3-gram-positive bacterial species, 3-gram negative bacterial species, 3-common bacterial species between aquatic and human, and 3-fungal species, and anticancer properties against breast cancer cells were performed. The results proved that the methanol extract has the highest antimicrobial, antifungal, antioxidant, and anticancer properties. After identifying the compounds of prepared methanol extract using GC/MS, polyvinylpyrrolidone nanofibers containing methanol extract of were prepared. The structure and characteristics of prepared nanofibers were confirmed and determined using FTIR, TGA, BET, SEM, flexural strength, compressive strength, and hydrophilicity. Synthesized polyvinylpyrrolidone nanofibers containing methanol extract of were subjected to antimicrobial properties on the strains studied in methanol extract of . The antimicrobial properties of synthesized polyvinylpyrrolidone nanofibers containing methanol extract of were compared. The results showed that synthesized polyvinylpyrrolidone nanofibers containing methanol extract of have the potential to introduction bioactive natural synthesis nanoparticles.
PubMed: 37954957
DOI: 10.3389/fchem.2023.1287870 -
Advanced Science (Weinheim,... Mar 2024Methane photooxidation into methanol offers a practical approach for the generation of high-value chemicals and the efficient storage of solar energy. However, the... (Review)
Review
Methane photooxidation into methanol offers a practical approach for the generation of high-value chemicals and the efficient storage of solar energy. However, the propensity for C─H bonds in the desired products to cleave more easily than those in methane molecules results in a continuous dehydrogenation process, inevitably leading to methanol peroxidation. Consequently, inhibiting methanol peroxidation is perceived as one of the most formidable challenges in the field of direct conversion of methane to methanol. This review offers a thorough overview of the typical mechanisms involved radical mechanism and active site mechanism and the regulatory methods employed to inhibit product peroxidation in methane photooxidation. Additionally, several perspectives on the future research direction of this crucial field are proposed.
PubMed: 38234232
DOI: 10.1002/advs.202306891 -
Food Science & Nutrition Oct 2023is rich in antioxidants such as polyphenols; contains bioactive compounds (flavonoids, terpenoids, and tannins) and is primarily composed of volatile oils, amides,...
is rich in antioxidants such as polyphenols; contains bioactive compounds (flavonoids, terpenoids, and tannins) and is primarily composed of volatile oils, amides, alkaloids, flavonoids, lignan, and coumarin. The antibacterial, antifungal, biofilm inhibition, antioxidant, hemolytic, and thrombolytic activities of , , and ethanol and methanol extracts at different concentrations (30%, 50%, and 80%) were determined. The antioxidant activity and content were measured as free radical scavenging assay (DPPH), total flavonoid content (TFC), and total phenolic content (TPC). Furthermore, hemolytic and thrombolytic analysis was carried out to determine toxicity. In antimicrobial assays, 80% methanol thyme extract showed highest (15.31 mm) antibacterial activity against , and 80% ethanol green tea extract showed optimal antibacterial activity against . Ethanol 30% green tea extract resulted in highest (26.61 mm) antifungal activity against . The maximum (54.73%) biofilm inhibition was resulted by methanol 50% thyme extract for . In antioxidant activity and content, methanol 50% green tea extract had highest (80.82%) antioxidant activity, whereas, ethanol 80% green tea extract had maximum (1474.55 mg CE/g DW) TFC and methanol 80% green tea extract had maximum (593.05 mg GAE/g) TPC. In toxicological assays, methanol 30% green tea extract had highest (25.28%) thrombolytic activity, and ethanol 80% tejphal extract had maximum (18.24%) hemolytic activity. This study has highlighted the significant antimicrobial, antioxidant, hemolytic, and thrombolytic activities of , , and extracts that could be beneficial to treat various diseases (cancer, diabetes, and respiratory diseases) and may be utilized as functional ingredient in the preparation of functional foods and drinks.
PubMed: 37823136
DOI: 10.1002/fsn3.3569