Kamal, R. M., A. M. El-Halawany, M. S. Hifnawy, A. M. Otify, W. G. Fahmy, N. M. Elhosseiny, A. S. Attia, B. M. Eltanany, L. Pont, F. Benavente, et al., "Targeting Acinetobacter baumannii lipase by coniferous species through metabolomics supported approach.", Scientific reports, vol. 15, issue 1, pp. 32649, 2025 Sep 23. Abstract

The opportunistic pathogen Acinetobacter baumannii is a particularly problematic nosocomial threat worldwide, leading to high morbidity and mortality rates due to its multiple resistance mechanisms, including the production of lipolytic enzymes. Herein, the aerial parts of three coniferous plants, Pinus canariensis C. Sm. (PC), Cupressus lusitanica Mill. (CL), and Cupressus arizonica Greene. (CA), were extracted and fractionated. Among these, the CA extract followed by CL and then PC, exhibited the highest inhibition of bacterial lipase activity, with half-maximal inhibitory concentration (IC) values of 1117 ± 87, 1278 ± 62, and 1926 ± 104 µg/mL, respectively. The methylene chloride fractions of CA and CL extracts exhibited the highest inhibition of bacterial lipase activity, with IC (940 ± 25 µg/mL and 1103 ± 155 µg/mL), respectively. The metabolite profile of the three extracts, along with their most active fractions, were determined using liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS). Interestingly, the metabolite profiles of CL extracts were established here for the first time. A total of 99 secondary metabolites from diverse classes were identified across all samples. Among these, four metabolites were isolated: 3,5-di-p-coumaroylquinic acid, epicatechin, cupressuflavone, and rutin. The biflavonoid cupressuflavone showed the lowest IC value (3812 ± 450 µg/mL). Additionally, partial least squares was applied to assess the key metabolites contributing to the differentiation of the studied bioactivity. Consequently, this study provides novel insights into the bioactivity potential of coniferous plants, demonstrating their value as a natural source of antivirulence agents against the A. baumannii lipase enzyme.

Elgahamy, A. A., A. H. El-Desoky, A. M. Otify, A. M. Elfishawy, and A. A. El-Beih, "Molecular networking derived from untargeted LC-MS/MS analysis to discover inhibitors of RANKL-induced osteoclastogenesis from Egyptian marine sponge-associated fungi.", Scientific reports, vol. 15, issue 1, pp. 27137, 2025 Jul 25. Abstract

In continuous search for RANKL induced osteoclastogenesis inhibitors, twenty-six fungal isolates were obtained from ten Red Sea marine sponges collected from Egypt and the ethyl acetate fractions of their cultures' methanol extracts were assessed in RAW264 macrophages. Active fractions were profiled via LC-MS/MS, followed by untargeted molecular networking, leading to the tentative identification of eight unreported compounds (1, A2, C1, C2, C4-C7), and thirteen known compounds. The two active fungi were identified and deposited in GenBank with accession numbers PQ423742 and PQ423748 for Aspergillus flavus and Cladosporium colombiae, respectively. Bioassay-guided isolation afforded two bisphenol diglycidic ethers, 1 and 2, and two diketopiperazines, 3 and 4 from A. flavus, while C. colombiae yielded cinnamic acid (5), two diketopiperazines (6 and 7), and altenuene (8). Structures were elucidated by NMR and mass spectroscopic analyses, which revealed 1 to be isolated for the first time from natural sources. Isolated compounds were biologically evaluated. Only 1 and 8 inhibited RANKL-induced formation of mature multinucleated osteoclasts with IC 57.14 and 38.35 µM, respectively, without cytotoxicity against RAW264 macrophages. The ADMET properties for 1 and 8 were predicated using SwissADME and pkCMS platforms. 8 showed superior solubility and lower toxicity than compound 1.

Farrag, A. F. S., A. M. Otify, A. I. M. Khedr, N. M. Misnan, A. Mediani, L. A. Wessjohann, M. G. Sharaf El-Din, and M. A. Farag, Unveiling metabolome heterogeneity in seed and husk from three cardamom species for quality control and valorization purposes of its waste products via NMR-based metabolomics in relation to in vitro biological effects, , vol. 480, pp. 143687, 2025. AbstractWebsite

NMR-based Metabolomics approach assessed phytochemical profile in seed and husk of three cardamom species: Elettaria cardamomum (green), Amomum subulatum (black), and Aframomum corrorima (white). NMR Spectroscopy identified 20 metabolites belonging to sugars, amino-, organic-, fatty acids, terpenes, and phenolics. Multivariate data analyses revealed distinct metabolic profiles among the 3 species, and further in seed versus husk. A. subulatum seed showed the highest sugar and amino acid levels, while E. cardamomum seed was richer in ω-3 fatty acids. Husk, especially from A. subulatum and E. cardamomum, contained high levels of phenolic acids. Compared to other cardamom taxa, A. corrorima exhibited lower levels of most chemicals. This study highlights the potential value of cardamom husk, particularly from A. subulatum and E. cardamomum species enriched in phenolic acids and terpenes known for their antioxidant and antimicrobial properties, for use as a food preservative. The antimicrobial and antioxidant activities were assessed through in vitro assays, revealing their potential for value-added applications in food preservation and therapeutic uses.

Ali, M. T., D. A. Al-Mahdy, A. M. Elfishawy, A. Salama, A. A. Al‐Karmalawy, and A. M. Otify, Phytochemical investigation, role in wound healing process and cytotoxicity of Sphagneticola trilobata: In vitro, in vivo and in silico approach, , vol. 342, pp. 119394, 2025. AbstractWebsite

Ethnopharmacological relevanceSphagneticola trilobata was traditionally used to alleviate wounds using topical plant preparations. The precise mechanism of the plant responsible for its wound healing effect are still unclear. Although the plant was reported to be cytotoxic, there is a lack of reported data regarding its cytotoxic impact on skin cancer.
Aim of the study
To in-depth investigate wound healing and cytotoxic effects of the plant extracts and its fractions by using different assays.
Materials and methods
In vitro scratch assay and in vivo full-thickness wound model were performed to evaluate the plant's wound healing activity. Enzyme-linked immunosorbent assay kits were employed to assess key skin parameters levels. The cytotoxicity was evaluated using in vitro Western Standard Time assay against skin carcinoma cells.
Results
Sphagneticola trilobata improved the wound closure percentage in both in vitro scratch assay and in vivo model by modulating inflammation, as evidenced by a reduction in tumor necrosis factor and mitogen activated protein kinase levels, and enhancing proliferation via elevating collagen, smooth muscle actin, and forkhead protein. The total extract's efficacy surpassed the effectiveness of reference ointment. The non-polar fraction outperformed the polar one in accelerating wound closure and improving the tested skin parameters levels. Trilobolide-6-O-isobutyrate displayed the best docking scores for the selected target receptors. Moreover, the non-polar fraction demonstrated the most powerful cytotoxic effect against skin cancer cells. A specific compound mixture (kaurenoic acid/grandiflorenic acid, 1 and 2) within this fraction exhibited stronger cytotoxicity.
Conclusions
Sphagneticola trilobata can be considered as a promising therapeutic agent in relieving wounds and combating skin cancer cell lines.

Atef, F., M. A. Abdelkawy, B. M. Eltanany, L. Pont, A. M. Fayez, M. F. Abdelhameed, F. Benavente, I. Y. Younis, and A. M. Otify, A comprehensive investigation of Clerodendrum Infortunatum Linn. using LC-QTOF-MS/MS metabolomics as a promising anti-alzheimer candidate, , vol. 15, issue 1, pp. 859, 2025. AbstractWebsite

Alzheimer’s disease (AD) poses a global health challenge, demanding innovative approaches for effective treatments. Clerodendrum infortunatum Linn. (Lamiaceae) is a shrub traditionally used as a medicinal plant to treat inflammation, skin diseases, and bronchitis. This study aims to identify the main bioactive metabolites in C. infortunatum using LC-QTOF-MS/MS and investigate its potential in protecting against cognitive decline in rats with scopolamine-induced AD disease. Metabolite profiling was performed on the methanol extract of the plant’s aerial parts using LC-QTOF-MS/MS. The inhibitory activity of the acetylcholinesterase enzyme was measured in vitro. To evaluate the cognitive effects, the methanol extract was orally administered at three doses (100, 200, and 400 mg/kg) to scopolamine-induced AD rats, and their cognitive functions were assessed using the novel object recognition test. Additionally, acetylcholinesterase enzyme activity, as well as the levels of acetylcholine, dopamine, noradrenaline, glutathione, malondialdehyde, tumor necrosis factor-α, interleukin-1β, and amyloid-β in the rat hippocampus, were measured using ELISA, followed by histopathological evaluation. A total of 79 metabolites, spanning various chemical classes, such as organic acids, phenolic acids, phenylpropanoids and phenylethanoids, flavonoids, coumarins, other phenolics, and fatty acids and their derivatives, were identified. The results showed that the extract promoted enhanced cognitive functions in the novel object recognition test. Scopolamine administration significantly altered the acetylcholinesterase enzyme activity and biomarker levels in the rat’s hippocampus. However, treatment with C. infortunatum at 200 and 400 mg/kg almost restored these neurotransmitter levels to normal, which was further confirmed by histopathological analysis. This study demonstrates the therapeutic potential of C. infortunatum in mitigating cognitive decline in AD, with its first metabolite profiling revealing a range of bioactive compounds. The extract improved cognitive function in scopolamine-induced AD rats, restored acetylcholinesterase activity, normalized neurotransmitter levels, and reduced oxidative stress and inflammation. These findings suggest that C. infortunatum is a promising candidate for the development of natural therapies targeting AD.

Abdel-Dayem, S. I. A., A. M. Otify, F. A. Iannotti, F. R. Saber, A. S. Moriello, S. Giovannuzzi, Ł. Świątek, A. Bonardi, P. Gratteri, K. Skalicka-Woźniak, et al., Damsin and neoambrosin: Two sesquiterpene lactones with affinity and different activity for PPAR and TRPA1 receptors, , vol. 154, pp. 108032, 2025. AbstractWebsite

Ambrosia maritima L. (family Asteraceae) is an annual herb widely distributed throughout the Mediterranean region and Africa. The herb is employed in folk medicine for the treatment of many ailments. Herein, we report a comprehensive investigation of the diverse biological potential of two sesquiterpene lactones, damsin and neoambrosin, isolated from Ambrosia maritima. 1D and 2D NMR and HR-ESI-MS/MS were employed to characterize the chemical structures of both compounds. In order to identify biological targets of both compounds we investigated their potential affinity for peroxisome proliferator-activated receptors (PPARs) and transient receptor potential (TRP) channels, which are pleiotropic classes of receptors implicated in essential functions of the body. This was investigated using a luciferase assay and a calcium fluorometric assay. A carbonic anhydrase inhibition assay was also performed using stopped flow CO2 hydrase spectrophotometric assay. Our analysis revealed that unlike damsin, neoambrosin showed a selective partial agonist effect on PPARγ receptors and TRPA1 channels. Its binding mode was investigated through in silico analysis. Both compounds showed no affinity for the tested carbonic anhydrases. Overall, our study details the chemical properties of neoambrosin and damsin and highlights neoambrosin as novel, cost-effective partial agonist of PPARɣ and TRPA1 receptors despite additional in vivo studies are needed to elucidate its biological and pharmacological properties.

Pecio, Ł., A. M. Otify, F. R. Saber, Y. A. El-Amier, M. E. Shalaby, S. Kozachok, A. K. Elmotayam, Ł. Świątek, A. Skiba, and K. Skalicka-Woźniak, "Iphiona mucronata (Forssk.) Asch. & Schweinf. A Comprehensive Phytochemical Study via UPLC-Q-TOF-MS in the Context of the Embryo- and Cytotoxicity Profiles", Molecules, vol. 27, issue 21, 2022. Abstract

Iphiona mucronata (Family Asteraceae) is widely distributed in the Eastern desert of Egypt. It is a promising plant material for phytochemical analysis and pharmacologic studies, and so far, its specific metabolites and biological activity have not yet been thoroughly investigated. Herein, we report on the detailed phytochemical study using UPLC-Q-TOF-MS approach. This analysis allowed the putative annotation of 48 metabolites belonging to various phytochemical classes, including mostly sesquiterpenes, flavonoids, and phenolic acids. Further, zebrafish embryotoxicity has been carried out, where 100 µg/mL extract incubated for 72 h resulted in a slow touch response of the 10 examined larvae, which might be taken as a sign of a disturbed peripheral nervous system. Results of in vitro testing indicate moderate cytotoxicity towards VERO, FaDu, and HeLa cells with CC50 values between 91.6 and 101.7 µg/mL. However, selective antineoplastic activity in RKO cells with CC50 of 54.5 µg/mL was observed. To the best of our knowledge, this is the first comprehensive profile of I. mucronata secondary metabolites that provides chemical-based evidence for its biological effects. A further investigation should be carried out to precisely define the underlying mechanisms of toxicity.

Ibrahim, R. M., B. M. Eltanany, L. Pont, F. Benavente, S. A. ElBanna, and A. M. Otify, "Unveiling the functional components and antivirulence activity of mustard leaves using an LC-MS/MS, molecular networking, and multivariate data analysis integrated approach", Food Research International, vol. 168, 2023. Abstract
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Otify, A. M., A. Serag, A. Porzel, L. A. Wessjohann, and M. A. Farag, "NMR Metabolome-Based Classification of Cymbopogon Species: a Prospect for Phyto-equivalency of its Different Accessions Using Chemometric Tools", Food Analytical Methods, vol. 15, no. 8: Springer, pp. 2095 – 2106, 2022. AbstractWebsite

Cymbopogon species are widely distributed worldwide and known for their high essential oil content with potential commercial and medicinal benefits justifying for their inclusion in food and cosmetics. Most species received scant characterization regarding their full complement of bioactive constituents necessary to explain their medicinal activities. In this study, the metabolite profiles of 5 Cymbopogon species, C. citratus, C. flexuosus, C. procerus, C. martini, and C. nardus, were characterized via NMR-based metabolomics. The results of 13 shoot accessions revealed the identification and quantification of 23 primary and secondary metabolites belonging to various compound classes. Multivariate analyses were used for species classification, though found not successful in discrimination based on geographical origin. Nevertheless, C. citratus was found particularly enriched in neral, geranial, (E)-aconitic acid, isoorientin, and caffeic acid as the major characterizing metabolites compared to other species, while an unknown apigenin derivative appeared to discriminate C. martini. The high essential oil and phenolic content in C. citratus emphasizes its strong antioxidant activity, whereas (E)-aconitic acid accounts for its traditional use as insecticide. This study affords the first insight into metabolite compositional differences among Cymbopogon species. Moreover, antimicrobial, insecticidal, antidiabetic, and antioxidant compounds were identified that can be utilized as biomarkers for species authentication. © 2022, The Author(s).

Otify, A. M., A. M. El-Sayed, C. G. Michel, and M. A. Farag, "Metabolites profiling of date palm (Phoenix dactylifera L.) commercial by-products (pits and pollen) in relation to its antioxidant effect: a multiplex approach of MS and NMR metabolomics", Metabolomics, vol. 15, no. 9: Springer New York LLC, 2019. AbstractWebsite

Introduction: Phoenix dactylifera L. (date palm) is one of the most valued crops worldwide for its economical and nutraceutical applications of its date fruit (pericarp). Currently date pits, considered as a waste product, is employed as coffee substitute post roasting. Whereas, pollen represents another valuable by-product used as a dietary supplement. Objectives: In this study, a large-scale comparative metabolomics approach was performed for the first characterization and standardization of date palm by-products viz., date pits and pollen. Moreover, roasting impact on date pit metabolite composition was also assessed. Methods: Metabolites profiling of pits and pollen was determined via a multiplex approach of UPLC–MS and NMR, coupled to multivariate analysis, in relation to its antioxidant activities. Results: Chemical analyses led to the identification of 67 metabolites viz., phenolic acids, flavonols, fatty acids, sphingolipids, steroids and saponins of which 10 are first time to be reported. The enrichment of steroids in date pollen accounts for its fertility promoting properties, whereas date pit was found a rich source for antioxidant polyphenols using metabolomics. © 2019, Springer Science+Business Media, LLC, part of Springer Nature.

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