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Methods and Protocols Nov 2023One approach to enhance the bioavailability and half-life of peptides in vivo is through N-methylation of one or more of the amino acids within the peptide sequence....
One approach to enhance the bioavailability and half-life of peptides in vivo is through N-methylation of one or more of the amino acids within the peptide sequence. However, commercially available Fmoc-N-Me-AA-OHs are limited and often expensive. In this study, a solid-phase synthesis method for Fmoc-N-Me-AA-OH was developed using a 2-chlorotrityl chloride (2-CTC) resin as a temporary protective group for the carboxylic acid strategy. Two strategies for the alkylation step were compared, employing either dimethyl sulfate or methyl iodide in the Biron-Kessler method. In this work we tested the protocol with two amino acids: Fmoc-Thr(tBu)-OH and Fmoc-βAla-OH. The first one is an alpha amino acid, very hindered and with the amine group directly influenced by the electronic effects of the carboxy group, whereas in Fmoc-βAla-OH, the presence of a methylene group weakens this influence due to the intervening carbon atoms. The desired amino acids, Fmoc-N-Me-Thr(tBu)-OH and Fmoc-N-Me-βAla-OH, were synthesized by both strategies with high yield and purity.
PubMed: 37987357
DOI: 10.3390/mps6060110 -
BMC Complementary Medicine and Therapies Nov 2023Anacyclus pyrethrum L. (Akarkara root), a valuable Ayurvedic remedy, is reported to exhibit various pharmacological activities. Akarkara root was subjected to...
Combinative effects of akarkara root-derived metabolites on anti-inflammatory and anti-alzheimer key enzymes: integrating bioassay-guided fractionation, GC-MS analysis, and in silico studies.
BACKGROUND
Anacyclus pyrethrum L. (Akarkara root), a valuable Ayurvedic remedy, is reported to exhibit various pharmacological activities. Akarkara root was subjected to bioassay-guided fractionation, to isolate its active constituents and discover their potential bioactivities, followed by computational analysis.
METHODS
The methanol extract and its fractions, methylene chloride, and butanol, were assessed for their antioxidant, anti-inflammatory, and anticholinergic potentials. The antioxidant activity was determined using DPPH, ABTS, FRAP, and ORAC assays. The in vitro anticholinergic effect was evaluated via acetyl- and butyryl-cholinesterase inhibition, while anti-inflammatory effect weas determined using COX-2 and 5-LOX inhibitory assays. The methylene chloride fraction was subjected to GC/MS analysis and chromatographic fractionation to isolate its major compounds. The inhibitory effect on iNOS and various inflammatory mediators in LPS-activated RAW 264.7 macrophages was investigated. In silico computational analyses (molecular docking, ADME, BBB permeability prediction, and molecular dynamics) were performed.
RESULTS
Forty-one compounds were identified and quantified and the major compounds, namely, oleamide (A1), stigmasterol (A2), 2E,4E-deca-2,4-dienoic acid 2-phenylethyl amide (A3), and pellitorine (A4) were isolated from the methylene chloride fraction, the most active in all assays. All compounds showed significant in vitro antioxidant, anticholinergic and anti-inflammatory effects. They inhibited the secretion of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) in activated RAW macrophages. The isolated compounds showed good fitting in the active sites of acetylcholinesterase and COX-2 with high docking scores. The ADME study revealed proper pharmacokinetics and drug likeness properties for the isolated compounds. The isolated compounds demonstrated high ability to cross the BBB and penetrate the CNS with values ranging from 1.596 to -1.651 in comparison with Donepezil (-1.464). Molecular dynamics simulation revealed stable conformations and binding patterns of the isolated compounds with the active sites of COX-2 and acetyl cholinesterase.
CONCLUSIONS
Ultimately, our results specify Akarkara compounds as promising candidates for the treatment of inflammatory and neurodegenerative diseases.
Topics: Antioxidants; Molecular Docking Simulation; Gas Chromatography-Mass Spectrometry; Acetylcholinesterase; Cyclooxygenase 2; Methylene Chloride; Plant Extracts; Anti-Inflammatory Agents; Anti-Bacterial Agents; Biological Assay; Cholinergic Antagonists
PubMed: 37978514
DOI: 10.1186/s12906-023-04210-6 -
Environmental Science & Technology Dec 2023Chloroform (CF) and dichloromethane (DCM) contaminate groundwater sites around the world but can be cleaned up through bioremediation. Although several strains of can...
Chloroform (CF) and dichloromethane (DCM) contaminate groundwater sites around the world but can be cleaned up through bioremediation. Although several strains of can reduce CF to DCM and multiple Peptococcaceae can ferment DCM, these processes cannot typically happen simultaneously due to CF sensitivity in the known DCM-degraders or electron donor competition. Here, we present a mixed microbial culture that can simultaneously metabolize CF and DCM and create an additional enrichment culture fed only DCM. Through genus-specific quantitative polymerase chain reaction, we find that grows while either CF alone or DCM alone is converted, indicating its involvement in both metabolic steps. Additionally, the culture was maintained for over 1400 days without the addition of an exogenous electron donor, and through electron balance calculations, we show that DCM metabolism would produce sufficient reducing equivalents (likely hydrogen) for CF respiration. Together, these results suggest intraspecies electron transfer could occur to continually reduce CF in the culture. Minimizing the addition of electron donor reduces the cost of bioremediation, and "self-feeding" could prolong bioremediation activity long after donor addition ends. Overall, understanding this mechanism informs strategies for culture maintenance and scale-up and benefits contaminated sites where the culture is employed for remediation worldwide.
Topics: Chloroform; Methylene Chloride; Biodegradation, Environmental; Halogenation; Peptococcaceae
PubMed: 37962431
DOI: 10.1021/acs.est.3c06686 -
International Journal of Molecular... Nov 2023The reactions of alkenes with phenyl--triflylimino-λ-iodane PhI=NTf () have been studied in different conditions. In methylene chloride, in the presence of...
The reactions of alkenes with phenyl--triflylimino-λ-iodane PhI=NTf () have been studied in different conditions. In methylene chloride, in the presence of -halosuccinimides, the products of mono and bis-triflamidation were obtained. In MeCN, the product of bromotriflamidation (with NBS) with solvent interception or of bis-triflamidation (with NIS) is formed. The reaction with -stilbene in acetonitrile with NBS gave rise to cyclization to 2-methyl-4,5-diphenyl-1-triflyl-4,5-dihydro-1-imidazole. In contrast, with NIS as an oxidant, both in CHCl and MeCN, the major product was 2,3-diphenyl-1-triflylaziridine formed in good yield. With NBS, aziridine is also formed but as a minor product, the major one being a mixture of diastereomers of the product of bromotriflamidation. The reaction of compound with vinylcyclohexane in methylene chloride affords the mixtures of regioisomers of the products of halotriflamidation, whereas in acetonitrile, the products of solvent interception and cyclization to the imidazoline are formed. A mechanism explaining the formation of all isolated products is proposed.
Topics: Oxidants; Molecular Structure; Solvents; Alkenes; Methylene Chloride; Acetonitriles
PubMed: 37958930
DOI: 10.3390/ijms242115947 -
Analytical Chemistry Nov 2023The effects of experimental repetitions and solvent extractors on the H NMR fingerprinting of yerba mate extracts, obtained from two genders and two light environments,...
The effects of experimental repetitions and solvent extractors on the H NMR fingerprinting of yerba mate extracts, obtained from two genders and two light environments, were analyzed in-depth by ANOVA-simultaneous component analysis (ASCA). Different solvents were used according to a mixture design based on ethanol, dichloromethane, and hexane and their combinations. The number of experimental repetitions significantly affected the ASCA results. Increasing repetitions led to decreases in the percentage effect variance values and an increase in the percentage residual variance. However, secondary sexual dimorphism, light availability, and their interaction effects became more significant with decreasing -values at or above the 95% confidence level. The choice of a solvent extractor significantly affects the chemical profile and can lead to distinct conclusions regarding the significance of effect values. Pure solvents yielded different conclusions about the significance of factorial design effects, with each solvent extracting unique metabolites and maximizing information for specific effects. However, the use of binary solvent mixtures, such as ethanol-dichloromethane, proved more efficient in extracting sets of compounds that simultaneously differentiate between different experimental conditions. The mixture design-fingerprint strategy provided satisfactory results expanding the range of extracted metabolites with high percentage of residual variances and low explained percentage effect variances in the ASCA models. Ternary and even higher-ordered mixtures could be good alternative extracting media for work-intensive procedures. Our study underscores the significance of experimental design and solvent selection in metabolomic analysis, improving the accuracy, robustness, and interpretability of metabolomic models, leading to a better understanding of the chemical composition and biological implications of plant extracts.
Topics: Ilex paraguariensis; Proton Magnetic Resonance Spectroscopy; Methylene Chloride; Plant Extracts; Solvents; Ethanol; Metabolome
PubMed: 37947492
DOI: 10.1021/acs.analchem.3c02592 -
Journal of Ethnopharmacology Feb 2024Clerodendrum infortunatum L. is commonly distributed in tropical and subtropical countries and is widely used in the indigenous systems of medicine including Ayurveda,...
ETHNOPHARMACOLOGICAL RELEVANCE
Clerodendrum infortunatum L. is commonly distributed in tropical and subtropical countries and is widely used in the indigenous systems of medicine including Ayurveda, Unani, and Homeopathy. The plant has important uses in the treatment of various inflammatory disorders which includes asthma, cough, chest complaints, pain, rheumatism, osteoarthritis, gastric ulcer, cancer, diabetes, skin diseases etc. AIM OF THE STUDY: Regarding the extensive healing properties of C. infortunatum in folk medicine, we aimed to explore the anti-inflammatory activity of the plant and analyze its bioactive components.
MATERIALS AND METHODS
Sequential extracts were prepared from C. infortunatum leaves using n-hexane, dichloromethane, ethyl acetate, methanol, ethanol and water. In vitro anti-inflammatory activity of these extracts were evaluated using membrane stabilization and protein anti-denaturation assays. Toxicity of the extracts was tested using brine shrimp lethality assay. GC-MS analysis has been carried out to analyze and characterize various bioactive constituents present in the plant.
RESULTS
Each of the tested extracts showed significant (p < 0.05) in vitro anti-inflammatory activity. Sequential ethyl acetate extract demonstrated the strongest membrane stabilization and protein anti-denaturation activities with IC values of 331.3, 308.3 and 279.2 μg/mL for heat and hypotonicity induced membrane stabilization and protein anti-denaturation assays, respectively. Sequential methanol extract also exhibited strong activity in heat and hypotonicity induced membrane stabilization and protein anti-denaturation assays with IC values of 371.5, 331.6 and 284.6 μg/mL, respectively. The extracts demonstrated a range of toxicity in the brine shrimp lethality assay with LC values of 51.4, 59.7, 47.3, 37.1, 68.1 and 77.3 μg/mL for n-hexane, dichloromethane, ethyl acetate, methanol, ethanol and water extracts respectively. The GC-MS analysis of direct n-hexane, ethyl acetate and methanol extracts detected 32 different compounds. The major compounds (> 5%) were phenol, 3,5-bis(1,1-dimethylethyl)-, hexadecanoic acid methyl ester, estragole, methyl stearate, lanosterol, 9-methoxybicyclo[6.1.0]nona-2,4,6-triene, benzenepropanoic acid, 3,5-bis(1,1-dimethylethyl) 4 hydroxy methyl ester, 22,23-dibromostigmasterol acetate, 3-methyl-4-(phenylthio)-2-prop-2-enyl-2,5-dihydrothiophene 1,1-dioxide, benzenemethanol, α-methyl-α-propyl- and 9-octadecenoic acid (Z)- methyl ester.
CONCLUSIONS
The findings of our study revealed that C. infortunatum is rich with diverse bioactive phytoconstituents and the scientific evidences support the anti-inflammatory potential of the plant.
Topics: Gas Chromatography-Mass Spectrometry; Plant Extracts; Clerodendrum; Methanol; Methylene Chloride; Anti-Inflammatory Agents; Ethanol; Water; Esters
PubMed: 37944873
DOI: 10.1016/j.jep.2023.117366 -
Carbohydrate Polymers Jan 2024Mushrooms contain chitin-glucan complex (CGC), a natural copolymer of chitin and glucan, and nanofibrillation enhances its applicability. Here, a novel method was used...
Mushrooms contain chitin-glucan complex (CGC), a natural copolymer of chitin and glucan, and nanofibrillation enhances its applicability. Here, a novel method was used to fabricate chitin-glucan nanofibers (CGNFs) from white button mushrooms. The first stage was to pretreat the raw mushroom using hot water and alkali to remove water-soluble glucans and alkali-soluble proteins, respectively, producing a CGC amenable to nanofibrillation. The second stage was nanofibrillation via esterification using acidic deep eutectic solvents (DESs) and subsequent ultrasonication. Five choline chloride-based DESs containing mono- or dicarboxylic acid were tested for the CGC esterification. DESs with strong dicarboxylic acids expedited nanofibrillation by homogeneously dispersing the solid CGC, swelling CGC fibrils, and facilitating acidity-dependent esterification leading to steric and electrostatic repulsions. One CGNF, namely CGNF_CCMnA, was characterized: it contained chitin and glucan at an approximate ratio of 8:2 and exhibited desirable properties as nanomaterials, including small diameter (11 nm) and high colloidal (zeta potential < -30 mV above pH 5.8) and thermal stability (T, 315 °C). CGNF_CCMnA was tested for the adsorption to methylene blue, revealing a maximum adsorption capacity of 82.58 mg/g. The proposed approach is an efficient and readily applicable method to fabricate various mushroom-derived safe CGNFs and to produce related nanomaterials.
PubMed: 37940284
DOI: 10.1016/j.carbpol.2023.121391 -
Acta Crystallographica. Section C,... Dec 2023The reaction of tris(pyridin-2-yl)amine with [CyRuCl] (Cy = p-isopropyltoluene or cymene) in refluxing diglyme led to the formation of cis-[RuCl{κ-(2-py)N}]·CHCl (1a)...
The reaction of tris(pyridin-2-yl)amine with [CyRuCl] (Cy = p-isopropyltoluene or cymene) in refluxing diglyme led to the formation of cis-[RuCl{κ-(2-py)N}]·CHCl (1a) after recrystallization from chloroform/pentane, or cis-dichloridobis[tris(pyridin-2-yl)amine-κN,N']ruthenium(II) dichloromethane disolvate, [RuCl(CHN)]·2CHCl or cis-[RuCl{κ-(2-py)N}]·2CHCl (1b). Treatment of 1a with one equivalent of silver(I) hexafluoridoantimonate in dichloromethane gave [RuCl{κ-(2-py)N}{κ-(2-py)N}][SbF]·CHCl (2a). Crystallization of 2a from chloroform provided chlorido[tris(pyridin-2-yl)amine-κN,N'][tris(pyridin-2-yl)amine-κN,N',N'']ruthenium(II) hexafluoridoantimonate chloroform monosolvate, [RuCl(CHN)][SbF]·CHCl or [RuCl{κ-(2-py)N}{κ-(2-py)N}][SbF]·CHCl (2b). Complex 2a reacted with a further equivalent of silver(I) hexafluoridoantimonate to give [Ru{κ-(2-py)N}][SbF] (3). The reaction of (2-py)N with [CyRuCl] in dichloromethane followed by treatment with excess sodium hexafluoridoantimonate gave the known complex [CyRuCl{κ-(2-py)N}][SbF] (4). Complex 2 is a rare example of a complex containing both κ- and κ-(2-py)N. Intramolecular π-stacking interactions determine the orientation of the free pyridyl in the κ complexes. An interesting encapsulation of methylene chloride hydrogen-bonded tetramers was noted in one case.
PubMed: 37929325
DOI: 10.1107/S2053229623009415 -
Heliyon Oct 2023Textile-based photocatalysts are the new materials that can be utilized as an effective sustainable solution for biochemical hazards. Hence, we aimed to develop a...
Textile-based photocatalysts are the new materials that can be utilized as an effective sustainable solution for biochemical hazards. Hence, we aimed to develop a sustainable, cost-effective, and facile approach for the fabrication of photocatalytic fabric using graphitic carbon nitride (g-CN) and ferric-based multifunctional nanocomposite. Bulk g-CN was prepared from urea by heating it at 500 °C for 2 h. The structure of ball-milled g-CN was engineered by doping with various amounts of iron (III) chloride hexahydrate solution (0.006 mol/L) and sintered at 90 °C for 24 h to prepare g-CN-nanosheets/α-FeO composites. These nanocomposites have potential avenues towards rational designing of g-CN for improved photocatalytic, antibacterial, and antiviral behavior. The prepared nanocomposite was characterized for its surface morphology, chemical composition, crystal structure, catalytic, antibacterial, and antiviral behavior. The fabrication of ferric doped g-CN nanocomposites was characterized by SEM, EDX, FTIR, and XRD analysis. The coated fabric nanocomposite was characterized for methylene blue dye degradation under visible light, antibacterial and antiviral behavior. The developed textile-based photocatalyst has been found with very good recyclability with photocatalytic degradation of dye up to 99.9 % when compared to conventional g-CN powder-based photocatalyst.
PubMed: 37886785
DOI: 10.1016/j.heliyon.2023.e20822 -
ACS Omega Oct 2023The effect of the ionic liquids (ILs) 1-methyl-3-tetradecylimidazolium chloride ([CMIM][Cl]), 1-dodecyl-3-methylimidazolium chloride ([CMIM][Cl]), and...
The effect of the ionic liquids (ILs) 1-methyl-3-tetradecylimidazolium chloride ([CMIM][Cl]), 1-dodecyl-3-methylimidazolium chloride ([CMIM][Cl]), and 1-decyl-methylimidazolium chloride ([CMIM][Cl]) on the structure of bovine serum albumin (BSA) was investigated by fluorescence spectroscopy, ultraviolet-visible (UV-vis) spectroscopy, small-angle X-ray scattering (SAXS), and molecular dynamics (MD) simulations. Concerning the fluorescence measurements, we observed a blue shift and a fluorescence quenching as the IL concentration increased in the solution. Such behavior was observed for all three studied imidazolium-based ILs, being larger as the number of methylene groups in the alkyl chain increased. UV-vis absorbance measurements indicate that even at relatively small IL/protein ratios, like 1:1 or 1:2, ([CMIM][Cl]) is able to change, at least partially, the sample turbidity. SAXS results agree with the spectroscopic techniques and suggest that the proteins underwent partial unfolding, evidenced by an increase in the radius of gyration () of the scattering particle. In the absence and presence of ([CMIM][Cl]) = 3 mM BSA increases from 29.1 to 45.1 Å, respectively. Together, these results indicate that the interaction of BSA with ILs is divided into three stages: the first stage is characterized by the protein in its native form. It takes place for protein/IL ≤ 1:2, and the interaction is predominantly due to the electrostatic forces provided by the negative charges on the surface of BSA and the cationic polar head of the ILs. In the second stage, higher IL concentrations induce the unfolding of the protein, most likely inducing the unfolding of domains I and III, in such a way that the protein's secondary structure is kept almost unaltered. In the last stage, IL micelles start to form, and therefore, the interaction with protein reaches a saturation point and free micelles may be formed. We believe that this work provides new information about the interaction of ILs with BSA.
PubMed: 37867681
DOI: 10.1021/acsomega.3c04188