LC-MS/MS metabolomics profiling of Glechoma hederacea L. methanolic extract; in vitro antimicrobial and in vivo with in silico wound healing studies on Staphylococcus aureus infected rat skin wound.

LC-MS/MS analysis of Glechoma hederacea L. methanolic extract (GHME), revealed the identification of 25 metabolites. Ursolic acid (1), 2α-hydroxyursolic acid or corosolic acid (2), 2ß-hydroxyursolic acid or epi-corosolic (3), luteolin 7-O-ß-D-glucopyranoside (4) and rosmarinic acid (5) were isolated and identified using spectroscopy. Antibacterial activity of GHME against multi drug resistance Staphylococcus aureus clinical isolates was measured. Minimum inhibitory concentrations (MICs) were ranged from 62.5 to 500 µg/ml. In vivo wound healing potential of 2%, and 5% GHME prepared hydrogels were criticized on Staphylococcus aureus infected wound rat model. 5% GHME prepared hydrogel treated group showed significant (p < 0.05) shrinkage of their colony forming unit/ml (CFU/ml) values in comparison with standard Fucidin. Meanwhile, wound closure associated with full re-epithelization and hair follicles proliferation was noticed after ten days of treatment. Finally, among the GHME isolated compounds, luteolin 7-O-ß-D-glucopyranoside (4) exhibited the highest molecular docking score (-9.6 kcal/mol) against matrix metalloproteinase-8 target (MMP-8).

Phytochemical Profile of the Ethanol Extract of Malvaviscus arboreus Red Flower and Investigation of the Antioxidant, Antimicrobial, and Cytotoxic Activities

Flowers are rich sources of bioactive antimicrobial, antioxidant, and anticancer components. This study aimed to determine the constituents of the ethanol extract of Malvaviscus arboreus red flower (ERF) by GC-MS analysis and HPLC identification of phenolic compounds and flavonoids, in addition to the 1HNMR fingerprint. The antimicrobial, antioxidant, and cytotoxic activities of the ERF were investigated. The GC-MS analysis revealed twenty-one components, while HPLC analysis revealed the presence of phenolic and flavonoid compounds. The ERF showed antifungal and antibacterial activity. The highest antibacterial activity was found against Vibrio damsela where a time-kill assay revealed a decline in the amount of viable V. damsela. For fungi, the highest activity was observed against Aspergillus terreus. Using the SRB test on HepG2, the anti-proliferative efficacy of the ERF was evaluated. Cell cycle analysis was utilized to determine autophagic cell death. The ERF prevented the proliferation of the HepG2 cell line with an IC50 of 67.182 µg/µL. The extract primarily promoted apoptosis in HepG2 cells by accumulating hypodiploid cells in the sub-G0/G1 phase, increased caspase 3/7 activity, and caused considerable autophagic cell death in apoptosis-deficient cells. Finally, the observed elevation of cancer cell death indicated that ERF had substantial anticancer potential against HepG2 cells.

In vitro and in silico ß-lactamase inhibitory properties and phytochemical profile of Ocimum basilicum cultivated in central delta of Egypt.

CONTEXT: Some studies reported the chemical content and antimicrobial properties of Ocimum basilicum L. (Lamiaceae), relevant to the ecological variations in some areas of Egypt and other countries, yet no research was conducted on the plant cultivated in the central delta region of Egypt. Also, no previous data reported on inhibition of ß-lactamases by O. basilicum. OBJECTIVE: To assess ß-lactamases inhibition by O. basilicum extracts and the individual constituents. MATERIALS AND METHODS: Dried aerial parts of O. basilicum were extracted by hydrodistillation for preparation of essential oil and by methanol for non-volatile constituents. Essential oil content and the methanol extract were analysed by GC-MS and UPLC-PDA-MS/MS, respectively. Methyl cinnamate was isolated and analysed by NMR. Broth microdilution method was used to investigate the antimicrobial against resistant clinical isolates of Escherichia coli identified by double disc synergy, combination disc tests and PCR. The most active oil content was further tested with a nitrocefin kit for ß-lactamase inhibition and investigated by docking. RESULTS: O. basilicum was found to contain methyl cinnamate as the major content of the essential oil. More interestingly, methyl cinnamate inhibited ESBL ß-lactamases of the type CTX-M. The in vitro IC50 using nitrocefin kit was 11.6 µg/mL vs. 8.1 µg/mL for clavulanic acid as a standard ß-lactamase inhibitor. DISCUSSION AND CONCLUSIONS: This is the first study to report the inhibitory activity of O. basilicum oil and methyl cinnamate against ß-lactamase-producing bacteria. The results indicate that methyl cinnamate could be a potential alternative for ß-lactamase inhibition.

Antibacterial activity and wound healing potential of Cycas thouarsii R.Br n-butanol fraction in diabetic rats supported with phytochemical profiling.

Patients with diabetes mellitus often suffer from chronic wounds due to wound healing impairment. Considering the increased prevalence of diabetes, this would predispose significant medical, economic, and social problems. These chronic wounds are frequently infected with pathogenic bacteria like Pseudomonas aeruginosa, which complicates the situation and makes the wound healing process more difficult. Therefore, there is a high need for therapeutic alternatives to the currently available treatments. Plants are vital sources of many bioactive compounds with multiple biological activities. We elucidated the wound healing possibility and antibacterial effect of Cycas thouarsii n-butanol fraction (CTBF) for the first time. Also, CTBF's phytochemical fingerprint was investigated using the LC-MS/MS technology. Interestingly, CTBF revealed antibacterial activity against P. aeruginosa isolates with minimum inhibitory concentrations range 16-128 µg/mL. Regarding the wound healing potential, we used in vivo experiment on diabetic rats. Remarkably CTBF caused a significant reduction (p 0.05) in the levels of forkhead box O1, matrix metalloproteinases 9, and chemokine (C-C motif) ligand 20. Additionally, it led to a substantial increase (p 0.05) in the level of transforming growth factor ß1. Moreover, CTBF improved the wound histological features by increasing the collagen area percentage. Regarding the immunohistochemical studies, CTBF resulted in a strong positive epidermal growth factor and a moderate positive caspase 9 immunoreaction in the epidermis and sebaceous glands of the wounds. Therefore, CTBF could be a promising source of bioactive compounds with wound healing and antibacterial activities. Finally, molecular docking was attempted using MOE software to investigate the binding mode of the major identified compounds in the matrix metalloproteinase 9 (MMP-9) receptor (PDB code: 1GKC).

Synthesis and characterization of polyvinyl alcohol/corn starch/linseed polyol-based hydrogel loaded with biosynthesized silver nanoparticles.

Nanocomposite hydrogel film was prepared from Polyvinyl alcohol [PVA], Corn Starch [CS], Linseed oil polyol [LP], and silver nanoparticles [NP]. LP was prepared by epoxidation and hydration of Linseed oil [LO]. IR and NMR supported the insertion of hydroxyl groups in LP by epoxide ring opening reaction at epoxidized LO. Silver NP were biosynthesized using aqueous leaves' extract from locally grown Ocimum forsskaolii Benth [LEO] plant. FTIR, XRD, UV and TEM confirmed the synthesis of NP (size 30 to 39 nm). Transparent and foldable hydrogel film resulted by blending the constituents (PVA, CS, LP and NP), crosslinking by glutaraldehyde, at room temperature, and showed expansion in water, different pH solutions, biodegradation and good antibacterial and antifungal activity against tested microbes. Linseed polyol influenced the structure, morphology, hydrophilicity, improved swelling ability and thermal stability and accelerated biodegradation of hydrogel films. NP were well adhered to LP globules that were embedded in PVA/CS matrix as strung set of beads (LP globules) decorated with black pearls (spherical NP). Silver NP conferred antimicrobial behavior to hydrogel film as observed by antimicrobial screening on different microbes. The results were encouraging and showed that such hydrogel films may find prospective applications in antimicrobial packaging.

Flavonoidal Constituents, Antioxidant, Antimicrobial, and Cytotoxic Activities of Dipterygium glaucum Grown in Kingdom of Saudi Arabia.

BACKGROUND: Dipterygium glaucum Decne. herb is one of the common traditional plants with multiple medicinal uses. OBJECTIVE: To isolate the major constituents and to investigate the antioxidant, antimicrobial, and cytotoxic activities of this herb. MATERIALS AND METHODS: Methanolic extract of D. glaucum herb was fractionated using n-hexane, dichloromethane, and n-butanol. Butanol fraction was chromatographed using column chromatography and preparative thin layer chromatography to isolate the major constituents. Isolated compounds were elucidated by means of spectroscopic methods, including 1D, 2D NMR (1H, 13C, DEPT, COSY, HSQC, HMBC, NEOSY) and MS analysis. Total phenolic content using Folin-Ciocalteu reagent and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay of the total methanolic extract were evaluated. Cytotoxic potential of both methanolic extract and butanol fraction was tested using a crystal violet viability assay. Antimicrobial activities of both extracts were investigated using diffusion agar technique. RESULTS: Apigenin 6, 8-di-C-glucopyranoside (vicenin-2), quercetin-3`-O-methyl-3-O-glucopyranoside, quercetin-3`-O-methyl-3-O-galactopyranoside, quercetin-3-O-ß-D-glucopyranoside, and quercetin-3-O-ß-D-galactopyranoside were isolated and elucidated. Total phenolic content was (83.89 mg gallic acid equivalent/g extract). The EC50 value of scavenging DPPH radical was 152.0 ± 2 mg/mL. Butanol fraction showed the highest cytotoxic activity against cervical and breast carcinoma cells (IC50 3.6 and 6.1 mg/mL, respectively). Both methanolic extract and butanol fraction showed wide spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria and some fungi. The highest activity was from methanolic extract against Enterococcus faecalis (83.25%) and against Candida tropicalis (77.03%) as compared to reference antibiotics. CONCLUSION: Data obtained from this study demonstrate that D. glaucum possesses significant antioxidant, cytotoxic, and antimicrobial activities which could be ascribed to its flavonoidal content. SUMMARY: Dipterygium glaucum Decne. herb is one of the common traditional plants with multiple medicinal uses in KSAFive flvonidal glycosides were isolated and elucidatedThis study demonstrated that D. glaucum possesses significant antioxidant, cytotoxic, and antimicrobial activities. Abbreviations used: KSA: Kingdom of Saudi Arabia; TLC: Thin Layer Chromatography; DPPH: 2,2`-diphenyl-1-picrylhydrazyl; EC50: Half maximal effective concentration; IC50: Half maximal inhibitory concentration; DMSO: dimethyl sulfoxide; NMR: Nuclear Magnetic Resonance; ESIMS: Electrospray ionization mass spectrometry; MeOH: Methyl alcohol.

Chemical and biological investigation of some secondary metabolites in Atriplex halimus growing in Egypt.

The n-BuOH-soluble fraction of the MeOH-CH2Cl2 (1:1) extract of the aerial parts of Egyptian Atriplex halimus L. yielded two new flavonol glycosides, designated as atriplexoside A (1) [3'-O-methylquercetin-4'-O-beta-apiofuranoside-3-O-(6"-O-alpha-rhamnopyranosyl-beta-glucospyranoside)] and atriplexoside B (2) [3'-O-methylquercetin-4'-O-(5"-O-beta-xylopyranosyl-beta-apiofuranoside)-3-O-(6"-O-alpha-rhamnopyranosyl-beta-glucopyranoside)], together with six known compounds: two phenolic glucosides (3, 4), one ecdysteroid (5), one megastigmane (6) and two methoxylated flavonoid glycosides (7, 8). The structures of the compounds were elucidated by detailed spectroscopic analysis, including HR-ESI-MS and 2D-NMR spectroscopic data. DPPH radical scavenging, antileishmanial and anti-multidrug resistance activities were investigated using the n-BuOH-soluble fraction as well as the isolated compounds. Compound 8 (5-O-methylquercetin-3-O-(6"-O-alpha-rhamnopyranosyl-beta-glucopyranoside) presented marked DPPH radical scavenging, weak antileishmanial and anti-multidrug resistance activity while the other tested compounds showed weaker activities.

In-vitro cytotoxic activities of the major bromophenols of the red alga Polysiphonia lanosa and some novel synthetic isomers.

Bioassay-guided fractionation was applied to the cytotoxic chloroform fraction of the red alga Polysiphonia lanosa. The major compounds of the most active fraction were identified using GLC-MS analysis as lanosol (1), methyl, ethyl, and n-propyl ethers of lanosol (1a, 1b, and 1c, respectively), and aldehyde of lanosol (2), although 1b appears to be an artifact arising during the fractionation procedure. These compounds and other known bromophenols were synthesized in addition to four novel isomers (3, 3a-c). The cytotoxic activities of all the synthetic compounds were determined against DLD-1 cells using the MTT assay. Compounds with IC(50) < 20 micromol were also tested against HCT-116 cells. Compound 3c (2,5-dibromo-3,4-dihydroxybenzyl n-propyl ether) was the most active compound against both cell lines (IC(50) = 1.72 and 0.80 micromol, respectively), and its effect on the cell cycle was studied using flow cytometry.