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Farag, M. A., E. U. Dokalahy, T. F. Eissa, I. M. Kamal, and A. Zayed, "Chemometrics-Based Aroma Discrimination of 14 Egyptian Mango Fruits of Different Cultivars and Origins, and Their Response to Probiotics Analyzed via SPME Coupled to GC-MS.", ACS omega, vol. 7, issue 2, pp. 2377-2390, 2022. Abstract

The present study investigated the volatile organic compounds (VOCs) in 14 Egyptian mango specimens collected from three different regions and of different cultivars (cvs). VOCs were extracted via solid-phase microextraction, followed by gas chromatography-mass spectrometry analysis. The results obtained for sesquiterpene hydrocarbons' qualitative abundance were represented by 28 peaks, whereas monoterpene hydrocarbons amounted for the highest levels in most of the mango cvs. Multivariate data analyses were employed for sample classification and identification of markers. Unsupervised principal component analysis revealed that "zebdia" cv from the three origins combined together being enriched in terpinolene. Moreover, supervised orthogonal partial least square-discriminant analysis identified β-terpinene and ()-geranylacetone in the premium "awees" cv. The impact of probiotic bacteria on mango juice aroma was further assessed revealing no potential changes in the composition. This study provides the first comprehensive insights into Egyptian mango aroma and reveals that the cv type overcomes the geographical origin in their aroma profile.

Farag, M. A., A. Meyer, and S. E. Ali, "Bleaching effect in Sarcophyton spp. soft corals-is there a correlation to their diterpene content?", Environmental science and pollution research international, vol. 28, issue 20, pp. 25594-25602, 2021. Abstract

Rising seawater temperature is one of the greatest threats to the persistence of coral reefs. While great efforts have been made to understand the metabolic costs of thermal acclimation, the exact roles of many secondary metabolites involved in the immediate response exhibited by soft corals remain largely unknown. Herein, an untargeted metabolomics approach using ultra-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC-MS) was employed to investigate thermal stress-induced modifications to the de novo synthesis of secondary metabolites in two soft coral species, Sarcophyton ehrenbergi and S. glaucum. Exposure to elevated temperature resulted in symbiont photoinhibition primarily via either damage to photosystem II (PSII) or the loss of algal symbionts during coral bleaching. This was suggested by a decrease in pulse amplitude modulated (PAM) measurements of corals incubated at different temperatures. Thermal stress was also found to impair the production of diterpenoid secondary metabolites in soft corals. Principally, reduction in the levels of a number of diterpenes, viz. sarcophytoxide and deoxysarcophytoxide, in heat stressed S. ehrenbergi and S. glaucum was observed indicative that thermal acclimation is energetically costly and will necessitate downstream changes in secondary metabolic pathways. Our data suggest that, while the host controls the production of ecologically important terpenes, when energetic contribution from the algal symbiont is reduced or absent as a result of a bleaching event, energy reserves may be insufficient to maintain the production of such energetically cost chemicals. This study provides for the first time a holistic assessment of secondary metabolite changes imposed in soft corals during exposure and acclimation to elevated temperatures.

Bakry, I. A., L. Yang, M. A. Farag, S. A. Korma, I. Khalifa, I. Cacciotti, N. I. Ziedan, J. Jin, Q. Jin, W. Wei, et al., "A Comprehensive Review of the Composition, Nutritional Value, and Functional Properties of Camel Milk Fat.", Foods (Basel, Switzerland), vol. 10, issue 9, 2021. Abstract

Recently, camel milk (CM) has been considered as a health-promoting icon due to its medicinal and nutritional benefits. CM fat globule membrane has numerous health-promoting properties, such as anti-adhesion and anti-bacterial properties, which are suitable for people who are allergic to cow's milk. CM contains milk fat globules with a small size, which accounts for their rapid digestion. Moreover, it also comprises lower amounts of cholesterol and saturated fatty acids concurrent with higher levels of essential fatty acids than cow milk, with an improved lipid profile manifested by reducing cholesterol levels in the blood. In addition, it is rich in phospholipids, especially plasmalogens and sphingomyelin, suggesting that CM fat may meet the daily nutritional requirements of adults and infants. Thus, CM and its dairy products have become more attractive for consumers. In view of this, we performed a comprehensive review of CM fat's composition and nutritional properties. The overall goal is to increase knowledge related to CM fat characteristics and modify its unfavorable perception. Future studies are expected to be directed toward a better understanding of CM fat, which appears to be promising in the design and formulation of new products with significant health-promoting benefits.

Ibrahim, H., A. Serag, and M. A. Farag, "Emerging analytical tools for the detection of the third gasotransmitter HS, a comprehensive review.", Journal of advanced research, vol. 27, pp. 137-153, 2021. Abstract

Background: Hydrogen sulfide (HS) is currently considered among the endogenously produced gaseous molecules that exert various signaling effects in mammalian species. It is the third physiological gasotransmitter discovered so far after NO and CO. HS was originally ranked among the toxic gases at elevated levels to humans. Currently, it is well-known that, in the cardiovascular system, HS exerts several cardioprotective effects including vasodilation, antioxidant regulation, inhibition of inflammation, and activation of anti-apoptosis. With an increasing interest in monitoring HS, the development of analysis methods should now follow.

Aim of review: This review stages special emphasis on the several analytical technologies used for its determination including spectroscopic, chromatographic, and electrochemical methods. Advantages and limitations with regards to the application of each technique are highlighted with special emphasis on its employment for HS measurement ., biofluids, tissues.

Key Scientific Concepts and important findings of Review: Fluorescence methods applied for HS measurement offer an attractive non-invasive and promising approach in addition to its selectivity, however they cannot be considered as H2S-specific probes. On the other hand, colorimetric assays are among the most common methods used for HS detection, albeit their employment HS measurement has not yet been possible . Separation techniques such as gas or liquid chromatography offer higher selectivity compared to direct spectrophotometric or fluorescence methods especially for suitable for endpoint HS measurements . plasma or tissue samples. Despite all the developed analytical procedures used for HS determination, the need for highly selective, much work should be devoted to resolve all the pitfalls of the current methods.

Ibrahim, H., A. Serag, and M. A. Farag, "Emerging analytical tools for the detection of the third gasotransmitter HS, a comprehensive review.", Journal of advanced research, vol. 27, pp. 137-153, 2021. Abstract

BACKGROUND: Hydrogen sulfide (HS) is currently considered among the endogenously produced gaseous molecules that exert various signaling effects in mammalian species. It is the third physiological gasotransmitter discovered so far after NO and CO. HS was originally ranked among the toxic gases at elevated levels to humans. Currently, it is well-known that, in the cardiovascular system, HS exerts several cardioprotective effects including vasodilation, antioxidant regulation, inhibition of inflammation, and activation of anti-apoptosis. With an increasing interest in monitoring HS, the development of analysis methods should now follow.

AIM OF REVIEW: This review stages special emphasis on the several analytical technologies used for its determination including spectroscopic, chromatographic, and electrochemical methods. Advantages and limitations with regards to the application of each technique are highlighted with special emphasis on its employment for HS measurement ., biofluids, tissues.

KEY SCIENTIFIC CONCEPTS AND IMPORTANT FINDINGS OF REVIEW: Fluorescence methods applied for HS measurement offer an attractive non-invasive and promising approach in addition to its selectivity, however they cannot be considered as H2S-specific probes. On the other hand, colorimetric assays are among the most common methods used for HS detection, albeit their employment HS measurement has not yet been possible . Separation techniques such as gas or liquid chromatography offer higher selectivity compared to direct spectrophotometric or fluorescence methods especially for suitable for endpoint HS measurements . plasma or tissue samples. Despite all the developed analytical procedures used for HS determination, the need for highly selective, much work should be devoted to resolve all the pitfalls of the current methods.

Yao, P., Y. Gao, J. Simal-Gandara, M. A. Farag, W. Chen, D. Yao, D. Delmas, Z. Chen, K. Liu, H. Hu, et al., "Litchi ( Sonn.): a comprehensive review of phytochemistry, medicinal properties, and product development.", Food & function, vol. 12, issue 20, pp. 9527-9548, 2021. Abstract

Since ancient times, litchi has been well recognized as a functional food for the management of various ailments. Many bioactives, including flavanoids, anthocyanins, phenolics, sesquiterpenes, triterpenes, and lignans, have been identified from litchi with a myriad of biological properties both and . In spite of the extensive research progress, systemic reviews regarding the bioactives of litchi are rather scarce. Therefore, it is crucial to comprehensively analyze the pharmacological activities and the structure-activity relationships of the abundant bioactives of litchi. Besides, more and more studies have focused on litchi preservation and development of its by-products, which is significant for enhancing the economic value of litchi. Based on the analysis of published articles and patents, this review aims to reveal the development trends of litchi in the healthcare field by providing a systematic summary of the pharmacological activities of its extracts, its phytochemical composition, and the nutritional and potential health benefits of litchi seed, pulp and pericarp with structure-activity relationship analysis. In addition, its by-products also exhibited promising development potential in the field of material science and environmental protection. Furthermore, this study also provides an overview of the strategies of the postharvest storage and processing of litchi.

Sheashea, M., J. Xiao, and M. A. Farag, "MUFA in metabolic syndrome and associated risk factors: is MUFA the opposite side of the PUFA coin?", Food & function, vol. 12, issue 24, pp. 12221-12234, 2021. Abstract

Omega-9 fatty acids represent some of the main mono-unsaturated fatty acids (MUFA) found in plant and animal sources. They can be synthesized endogenously in the human body, but they do not fully provide all the body's requirements. Consequently, they are considered as partially essential fatty acids. MUFA represent a healthier alternative to saturated animal fats and have several health benefits, including the prevention of metabolic syndrome (MetS) and its complications. This review concentrates on the major MUFA pharmacological activities in the context of MetS management, including alleviating cardiovascular disease (CVD) and dyslipidemia, central obesity, non-alcoholic fatty liver disease (NAFLD), and type 2 diabetes mellitus (T2DM). The beneficial effects of MUFA for CVD were found to be consistent with those of polyunsaturated fatty acids (PUFA) for the alleviation of systolic and diastolic blood pressure and high low density lipoprotein cholesterol (LDLc) and triacylglcerol (TAG) levels, albeit MUFA had a more favorable effect on decreasing night systolic blood pressure (SBP). To reduce the obesity profile, the use of MUFA was found to induce a higher oxidation rate with a higher energy expenditure, compared with PUFA. For NAFLD, PUFA was found to be a better potential drug candidate for the improvement of liver steatosis in children than MUFA. Any advantageous outcomes from using MUFA for diabetes and insulin resistance (IR) compared to using PUFA were found to be either non-significant or resulted from a small number of meta-analyses. Such an increase in the number of studies of the mechanisms of action require more clinical and epidemiological studies to confirm the beneficial outcomes, especially over a long-term treatment period.

Chen, Z., M. A. Farag, Z. Zhong, C. Zhang, Y. Yang, S. Wang, and Y. Wang, "Multifaceted role of phyto-derived polyphenols in nanodrug delivery systems.", Advanced drug delivery reviews, vol. 176, pp. 113870, 2021. Abstract

As naturally occurring bioactive products, several lines of evidence have shown the potential of polyphenols in the medical intervention of various diseases, including tumors, inflammatory diseases, and cardiovascular diseases. Notably, owing to the particular molecular structure, polyphenols can combine with proteins, metal ions, polymers, and nucleic acids providing better strategies for polyphenol-delivery strategies. This contributes to the inherent advantages of polyphenols as important functional components for other drug delivery strategies, e.g., protecting nanodrugs from oxidation as a protective layer, improving the physicochemical properties of carbohydrate polymer carriers, or being used to synthesize innovative functional delivery vehicles. Polyphenols have emerged as a multifaceted player in novel drug delivery systems, both as therapeutic agents delivered to intervene in disease progression and as essential components of drug carriers. Although an increasing number of studies have focused on polyphenol-based nanodrug delivery including epigallocatechin-3-gallate, curcumin, resveratrol, tannic acid, and polyphenol-related innovative preparations, these molecules are not without inherent shortcomings. The active biochemical characteristics of polyphenols constitute a prerequisite to their high-frequency use in drug delivery systems and likewise to provoke new challenges for the design and development of novel polyphenol drug delivery systems of improved efficacies. In this review, we focus on both the targeted delivery of polyphenols and the application of polyphenols as components of drug delivery carriers, and comprehensively elaborate on the application of polyphenols in new types of drug delivery systems. According to the different roles played by polyphenols in innovative drug delivery strategies, potential limitations and risks are discussed in detail including the influences on the physical and chemical properties of nanodrug delivery systems, and their influence on normal physiological functions inside the organism.

El-Shamy, S., and M. A. Farag, "Novel trends in extraction and optimization methods of bioactives recovery from pomegranate fruit biowastes: Valorization purposes for industrial applications.", Food chemistry, vol. 365, pp. 130465, 2021. Abstract

Pomegranate biowastes present potential economic value worldwide owing to their several health benefits mediated by a complex mixture of unique bioactives. The exploitation of these bioactives has motivated the exploration of eco-friendly, efficient, and cost-effective extraction techniques to maximize their recovery. The current review aims to provide updated technical information about bioactives extraction mechanisms from pomegranate wastes (seeds and peel), their advantages and disadvantages, and factors towards optimization. A comparative overview of the modern green extraction techniques viz., supercritical fluid extraction, ultrasound-assisted extraction, microwave-assisted extraction, pressurized liquid extraction, and eutectic solvent mixture as alternatives to conventional extraction methods for seeds and peel is presented. Approaches focused on biowastes modification for properties improvement are also discussed. Such comprehensive review shall provide the best valorization practices of pomegranate biowastes and its application in food and non-food areas focusing on original methods, innovation, protocols, and development to be considered for other fruit biowastes.

Elshahed, M. S., A. Miron, A. C. Aprotosoaie, and M. A. Farag, "Pectin in diet: Interactions with the human microbiome, role in gut homeostasis, and nutrient-drug interactions.", Carbohydrate polymers, vol. 255, pp. 117388, 2021. Abstract

Pectins are a part of daily diet as well as food additives that are indigestible polysaccharides by human enzymes, however, they can be easily degraded by gut bacteria with the production of short chain fatty acids (SCFAs). Knowledge of pectin gut homeostasis and further how pectin affect gut bacterial communities is insufficient and limited. This review focuses on providing the whole story of how pectin functions as prebiotics in the gut. Understanding the interplay between functional and immunological responses inside animal or human gut as influenced by pectin in diets is provided. The interaction between pectin and gut microbiota is presented from both sides, in terms of how pectin affects gut microbiome and or the fermentation products produced in response by gut bacteria. This knowledge can be used to define preferred dietary pectins, targeting beneficial bacteria, and favoring balanced microbiota communities in the gut to maximize pectins' health benefits.

Serag, A., Z. Shakkour, A. M. Halboup, F. Kobeissy, and M. A. Farag, "Sweat metabolome and proteome: Recent trends in analytical advances and potential biological functions.", Journal of proteomics, vol. 246, pp. 104310, 2021. Abstract

Metabolome and proteome profiling of biofluids, e.g., urine, plasma, has generated vast and ever-increasing amounts of knowledge over the last few decades. Paradoxically, omics analyses of sweat, one of the most readily available human biofluids, have lagged behind. This review capitalizes on the current knowledge and state of the art analytical advances of sweat metabolomics and proteomics. Moreover, current applications of sweat omics such as the discovery of disease biomarkers and monitoring athletic performance are also presented in this review. Another area of emerging knowledge that has been highlighted herein lies in the role of skin host-microbiome interactions in shaping the sweat metabolite-protein profiles. Discussion of future research directions describes the need to have a better grasp of sweat chemicals and to better understand how they function as aided by advances in omics tools. Overall, the role of sweat as an information-rich biofluid that could complement the exploration of the skin metabolome/proteome is emphasized.

Zayed, A., M. T. Badawy, and M. A. Farag, "Valorization and extraction optimization of Citrus seeds for food and functional food applications.", Food chemistry, vol. 355, pp. 129609, 2021. Abstract

Valorization of food byproducts has attracted recently considerable attention. Citrus fruits provide considerable non-edible residues reach 80% in juice production. They are considered agri-wastes to comprise peel, pulp and seeds. Previous investigations have focused on peel and pulp to recover value-added products. The review presents for the first-time phytochemical composition of Citrus seeds' products, i.e., oil and extracts. Fatty acids, phytosterols and tocopherols amounted as the major bioactives in Citrus seeds, in addition to limonoids, dietary fibers and flavonoids. Besides their nutritional values, these chemicals have promising applications including production of biodiesel, food enhancers and antioxidants, especially from mandarin and grapefruit seeds. Optimum conditions of the different Citrus seeds' valorization are discussed to improve extraction yield and lessen environmental hazards of solvent extraction. This review presents the best utilization practices for one of the largest cultivated fruit seeds worldwide and its different applications.

Emad, A. M., S. F. Ali, E. A. Abdel-Rahman, M. R. Meselhy, M. A. Farag, S. S. Ali, and E. A. Abdel-Sattar, "Anti-inflammatory and antioxidant effects of Apium graveolens L. extracts mitigate against fatal acetaminophen-induced acute liver toxicity.", Journal of food biochemistry, pp. e13399, 2020. Abstract

In the present work, antioxidant activity, total phenolics (TP), and total flavonoids (TF) contents of aqueous and methanol extracts of celery were determined, in addition to untargeted metabolites profiling its methanol celery root extract (MCRE) via UPLC-MS. Although MCRE exhibited the lowest TPC and TFC levels, it presented the most potential hydroxyl radical quenching effect using electron paramagnetic resonance spin trapping technique. Treatment of Acetaminophen-induced hepatotoxicity (AAH) rats with MCRE lowered serum levels of AST, ALT, ALP, TNF-α, and IL-1β significantly. Additionally, MCRE significantly increased total antioxidant capacity (TAC) and glutathione (GSH) levels relative to AAH rats. Strikingly, Kaplan-Meier survival analysis of all groups revealed a 100% prevention of acetaminophen-induced mortality of rats by MCRE pretreatment (100 mg/kg/day). MCRE prevented AAH-associated severe weight loss and elicited normal behavior in the rescued rats. Our results suggest that pretreatment with MCRE can mitigate against overdosed acetaminophen-induced acute liver failure and warrant further investigations on the potential of postinjury intervention. PRACTICAL APPLICATIONS: Acetaminophen-induced hepatotoxicity (AAH) accounts for alerting numbers of overdose-related acute liver failure and liver transplant cases with increased morbidity and mortality rates. Currently proposed mechanisms implicate mitochondria-mediated oxidative stress and inflammation in the pathogenesis of AAH, which underline current interventions employing antioxidants to combat liver damage by over-dosed acetaminophen. The present work uncovers potent protective effects of some celery extracts (and their fractions) against acetaminophen-induced oxidative stress and inflammation. Treatment of rats with fatal liver injury with methanol extract of celery root significantly reduced secretion of liver enzymes and markedly decreased inflammatory as well as oxidative stress markers in these animals. This, in turn, rescued challenged rats exposed to fatal doses of acetaminophen completely, which establishes methanol extracts of celery roots as effective therapeutic intervention against AAH. The antioxidant capacity of the extracts was determined using EPR technique, and the secondary metabolites related to antioxidant activity were characterized via UPLC-MS.

Farag, M. A., N. Hegazi, E. Dokhalahy, and A. R. Khattab, "Chemometrics based GC-MS aroma profiling for revealing freshness, origin and roasting indices in saffron spice and its adulteration.", Food chemistry, vol. 331, pp. 127358, 2020. Abstract

Saffron, stigmas of Crocus sativus, is one of the most precious spices used as food colorant and flavoring agent. Due to its scarce source and high cost, it is liable to fraudulent admixture with allied plants "safflower and calendula". In this study, gas chromatography-mass spectrometry (GC-MS) was employed to determine authenticity, adulterants detection, and to assess the roasting impact on its aroma. A total of 93 volatiles were identified belonging to different classes viz. aldehydes, alcohols, ketones, aliphatic hydrocarbons, aromatics, mono-and sesquiterpenes, oxides/ethers and pyrans/furans. Principle component analysis (PCA) identified safranal and 2-caren-10-al as discriminatory volatile markers of saffron from its allied flowers, later found enriched in estragole, β-caryophyllene and eugenol. PCA model also revealed markers for freshly dried versus long-stored saffron, with ketoisophorone as freshness marker versus safranal as an ageing indicator. Safranal was further identified as a marker to distinguish saffron from safflower, whereas calendula aroma was predominated by monoterpene hydrocarbons.

Khattab, A. R., and M. A. Farag, "Current status and perspectives of xanthones production using cultured plant biocatalyst models aided by tools for its optimization.", Critical reviews in biotechnology, vol. 40, issue 3, pp. 415-431, 2020. Abstract

With an increasing interest in exploring the biocatalyst potential for the production of natural products (NP), research in NP biotechnology now follows. Xanthones constitute an important class of phytochemicals exhibiting a myriad of bioactivities and potential for interacting with multimolecular targets, and xanthones are considered to be privileged phytochemical scaffolds in medicinal chemistry. Recent advances in xanthones biosynthetic pathways are outlined and their metabolic engineering. This review summarizes the state-of-the-art processes for the biotechnological production of xanthones, its limitations, and potential improvements. The tissue culturing methods employed for the production of xanthones from different species of plant are reviewed addressing differences in xanthone classes. This review sheds the light of computational approaches, , response surface methodology (RSM) and artificial neural networks (ANN), for optimizing the culture performance and improving xanthones production.

Shakour, Z. A. T., N. m Fayek, and M. A. Farag, "How do biocatalysis and biotransformation affect dietary flavonoids chemistry and bioactivity? A review.", Critical reviews in biotechnology, vol. 40, issue 5, pp. 689-714, 2020. Abstract

Flavonoids, especially flavanones, flavones and polymethoxyflavones (PMFs) are distinctive bioactive compounds of fruits. peel and juice by-products potentially represent rich sources of these flavonoids that exhibit a myriad of biological activities both in and systems. Recently, much attention has been made toward biotransformation processes as a promising tool for the structural modification of natural products to be used in the drug and food industries along with its role in solving pollution problems related to by-products disposal. In this article, we present a state of the art review on both and biotransformation processes of juice and waste carried out by microorganisms, plant cell cultures, animal and human liver microsomes targeting its flavonoids composition. Such review highlights the main metabolic pathways for enzymatic and microbial reactions involved in these processes and suggest for reactions that need to be more capitalized for a wider application in industrial bioprocesses of by-products. Biotransformation and biocatalysis applications included employment of citrus by-products enriched in flavonoids as a low-cost, economical and natural sources of sugar substitutes, antifungal, anticancer drugs, hydrolyzable enzymes, probiotics and flavonoid aglycones. Further, biological effects of the biotransformed metabolites are discussed in relation to its parent compound highlighting potentials and or any limitations for each reaction type. Applications covered in biotransformation include for nutraceutical/food and cosmetics industries.

Farag, M. A., S. A. Jomaa, A. Abd El-Wahed, and A. H. R. El-Seedi, "The Many Faces of Kefir Fermented Dairy Products: Quality Characteristics, Flavour Chemistry, Nutritional Value, Health Benefits, and Safety.", Nutrients, vol. 12, issue 2, 2020. Abstract

Kefir is a dairy product that can be prepared from different milk types, such as goat, buffalo, sheep, camel, or cow via microbial fermentation (inoculating milk with kefir grains). As such, kefir contains various bacteria and yeasts which influence its chemical and sensory characteristics. A mixture of two kinds of milk promotes kefir sensory and rheological properties aside from improving its nutritional value. Additives such as inulin can also enrich kefir's health qualities and organoleptic characters. Several metabolic products are generated during kefir production and account for its distinct flavour and aroma: Lactic acid, ethanol, carbon dioxide, and aroma compounds such as acetoin and acetaldehyde. During the storage process, microbiological, physicochemical, and sensory characteristics of kefir can further undergo changes, some of which improve its shelf life. Kefir exhibits many health benefits owing to its antimicrobial, anticancer, gastrointestinal tract effects, gut microbiota modulation and anti-diabetic effects. The current review presents the state of the art relating to the role of probiotics, prebiotics, additives, and different manufacturing practices in the context of kefir's physicochemical, sensory, and chemical properties. A review of kefir's many nutritional and health benefits, underlying chemistry and limitations for usage is presented.

El-Sabagh, O. A., S. A. El-Toumy, R. Mounir, M. A. Farag, and E. A. Mahrous, "Metabolite profiles of Pulicaria crispa and P. incisa in relation to their in-vitro/ in-vivo antioxidant activity and hepatoprotective effect: A comparative mass spectrometry-based metabolomics.", Journal of pharmaceutical and biomedical analysis, pp. 113804, 2020. Abstract

Plants of the genus Pulicaria (family: Asteraceae) are widely used in central Asia and the Middle East for treatment of different human diseases. Ultra performance liquid chromatography coupled to high resolution mass spectrometry (UPLC/MS) was utilized to establish the metabolic profiles of two Pulicaria species: P. crispa and P. incisa. 122 metabolites were identified including flavonoids (37), phenolic acids (22), sesquiterpenes (17), diterpenes (7), and fatty acids (27), with enrichment in methoxylated flavonoids (20), caffeoylquinic acid conjugates (14) xanthane sesquiterpenes (9) and hydroxylated fatty acids (20) in both Pulicaria species. The metabolite profile of P. incisa was characterized by the presence of tri- and tetra-methoxylated flavonoids while xanthane sesquiterpenes were the main chemical markers of P. crispa. Additionally, a novel sesquiterpene acid (dihydropulicaric acid) was annotated in both species based on its MS fragments. Antioxidant activity for P. crispa and P. incisa methanol extracts was assessed in vitro based on DPPH and ABTS assays and further in vivo using chlorpromazine intoxicated rat model. Results revealed that P. incisa extract was more effective in inhibiting both DPPH and ABTS free radicals (IC 0.36 and 0.52 mg/mL, respectively) than P. crispa (IC 0.51 and 0.73 mg/mL). In the animal model, antioxidant activity of P. incisa (20 mg/kg/day) was also slightly higher causing a 55 % reduction in MDA levels and 65 % increase in GSH activity compared to untreated animals. Furthermore, both extracts showed a hepatoprotective effect as revealed by improvement in levels of serum biomarkers of liver functions: total bilirubin, alanine transaminase (ALT) and aspartate transaminase (AST) comparable to silymarin at 25 mg/kg/day. These findings were also supported by the preserved integrity of the hepatic tissues of animals receiving either extracts at a dose of 20 mg/kg b.wt. The present study reveals for the potential antioxidant and hepatoprotective effects for Pulicaria in relation to its bioactive metabolites.

Baky, M. H., M. A. Farag, and D. M. Rasheed, "Metabolome-Based Analysis of Herbal Cough Preparations Headspace Solid-Phase Microextraction GC/MS and Multivariate Data Analyses: A Prospect for Its Essential Oil Equivalency.", ACS omega, vol. 5, issue 48, pp. 31370-31380, 2020. Abstract

Liquid cough preparations containing essential oils pose a challenge for isolating and quantifying their volatile components from such a complex matrix enriched with nonvolatile constituents and excipients. This study aims to develop a strategy integrating QC analysis of seven natural cough preparations in the Egyptian market and to assess volatile variation among the preparations using multivariate data analyses. Cough preparations were subjected to headspace solid-phase microextraction (HS-SPME) for determination of their essential oil composition mediating for their actions and to assess volatile differences among them. HS-SPME is a suitable technique for sample preparation that allows for extraction and enrichment of volatiles from complex nonvolatile matrices and their direct desorption into the gas chromatography analytical system. A total of 88 volatile components were identified belonging to seven classes, . aromatics, aliphatic hydrocarbons, mono/sesquiterpene hydrocarbons, and oxygenated mono/sesquiterpenes. Oxygenated monoterpenes, ., menthol, cineole, thymol, and ()-anethole, were the major volatiles identified in five cough preparations (79.5-98.6%), whereas aromatics, chiefly cinnamate derivatives, constituted the second class amounting for 50.5 and 27.4% in the other two cough preparations. Meaningful results regarding the products' efficacy and safety were extrapolated from this analytical procedure, where artificial preservatives (parabens) were detected in five cough preparations. This study established an efficient strategy for exploring volatile profiling and defining different markers among the different cough preparations. Additionally, authenticity of listed herbal ingredients in the cough preparations was also confirmed in certain preparations, while other formulations failed to show representative volatile components. Volatile variation among preparations was assessed using multivariate data analyses in an attempt to prioritize cough preparations for usage, suggesting the preference of Bronchicum and Babetone among examined cough products.

Farag, M. A., A. AbdelWareth, I. E. Sallam, M. El Shorbagi, N. Jehmlich, K. Fritz-Wallace, S. Serena Schäpe, U. Rolle-Kampczyk, A. Ehrlich, L. A. Wessjohann, et al., "Metabolomics reveals impact of seven functional foods on metabolic pathways in a gut microbiota model.", Journal of advanced research, vol. 23, pp. 47-59, 2020. Abstract

Functional food defined as dietary supplements that in addition to their nutritional values, can beneficially modulate body functions becomes more and more popular but the reaction of the intestinal microbiota to it is largely unknown. In order to analyse the impact of functional food on the microbiota itself it is necessary to focus on the physiology of the microbiota, which can be assessed in a whole by untargeted metabolomics. Obtaining a detailed description of the gut microbiota reaction to food ingredients can be a key to understand how these organisms regulate and bioprocess many of these food components. Extracts prepared from seven chief functional foods, namely green tea, black tea, (prickly pear, cactus pear), black coffee, green coffee, pomegranate, and sumac were administered to a gut consortium culture encompassing 8 microbes which are resembling, to a large extent, the metabolic activities found in the human gut. Samples were harvested at 0.5 and 24 h post addition of functional food extract and from blank culture in parallel and analysed for its metabolites composition using gas chromatography coupled to mass spectrometry detection (GC-MS). A total of 131 metabolites were identified belonging to organic acids, alcohols, amino acids, fatty acids, inorganic compounds, nitrogenous compounds, nucleic acids, phenolics, steroids and sugars, with amino acids as the most abundant class in cultures. Considering the complexity of such datasets, multivariate data analyses were employed to classify samples and investigate how functional foods influence gut microbiota metabolisms. Results from this study provided a first insights regarding how functional foods alter gut metabolism through either induction or inhibition of certain metabolic pathways, i.e. GABA production in the presence of higher acidity induced by functional food metabolites such as polyphenols. Likewise, functional food metabolites ., purine alkaloids acted themselves as direct substrate in microbiota metabolism.

Farag, M. A., N. M. Hegazi, and M. S. Donia, "Molecular networking based LC/MS reveals novel biotransformation products of green coffee by ex vivo cultures of the human gut microbiome.", Metabolomics : Official journal of the Metabolomic Society, vol. 16, issue 8, pp. 86, 2020. Abstract

INTRODUCTION: Unroasted green coffee bean is an increasingly popular beverage and weight loss supplement that contains higher levels of chlorogenic acid derivatives and lower alkaloid levels than roasted beans. Nonetheless, how the gut microbiome metabolizes green coffee constituents has not been studied.

OBJECTIVES: To identify possible biotransformation products of green coffee extract by the human gut microbiome, and the potential implications of this process on its biological effects or fate inside the body.

METHODS: Molecular networking via the GNPS platform was employed for the visualization of green coffee metabolite profiles acquired using LC-tandem mass spectrometry post-incubation with an ex vivo culture of the human gut microbiome.

RESULTS: 36 Metabolites were annotated including four unreported alkyl cinnamate esters in green coffee along with six novel biotransformation products.

CONCLUSION: Our finding reveals new biotransformation products of cinnamate esters by the gut microbiome mediated via oxidative reactions such as dehydrogenation and hydroxylation, along with methylation, decarboxylation, and deglycosylation. These findings reveal potential interactions between the gut microbiome and green coffee constituents, and paves the way towards studying the effects of these interactions on both microbiome and the human host.

Ramadan, N. S., L. A. Wessjohann, A. Mocan, D. C. Vodnar, N. H. El-Sayed, S. A. El-Toumy, D. A. Mohamed, Z. A. Aziz, A. Ehrlich, and M. A. Farag, "Nutrient and Sensory Metabolites Profiling of L. (Starfruit) in the Context of Its Origin and Ripening Stage by GC/MS and Chemometric Analysis.", Molecules (Basel, Switzerland), vol. 25, issue 10, 2020. Abstract

L. is a tropical tree with edible fruit that grows at different climatic conditions. Despite its nutritive value and reported health benefits, it is a controversial fruit owing to its rich oxalate content. The present study aimed at investigating aroma and nutrient primary metabolites distribution in fruits grown in Indonesia, Malaysia (its endemic origin) versus Egypt, and at different ripening stages. Two techniques were employed to assess volatile and non-volatile metabolites including headspace solid-phase micro-extraction (HS-SPME) joined with gas chromatography coupled with mass-spectrometry (GC-MS) and GC-MS post silylation, respectively. Twenty-four volatiles were detected, with esters amounting for the major class of volatiles in Egyptian fruit at ca. 66%, with methyl caproate as the major component, distinguishing it from other origins. In contrast, aldehydes predominated tropically grown fruits with the ether myristicin found exclusively in these. Primary metabolites profiling led to the identification of 117 metabolites viz. sugars, polyols and organic acids. Fructose (38-48%) and glucose (21-25%) predominated sugar compositions in ripe fruits, whereas sorbitol was the major sugar alcohol (2.4-10.5%) in ripe fruits as well. Oxalic acid, an anti-nutrient with potential health risks, was the major organic acid detected in all the studied fruits (1.7-2.7%), except the Malaysian one (0.07%). It increases upon fruit ripening, including considerable amounts of volatile oxalate esters detected via SPME, and which must not be omitted in total oxalate determinations for safety assessments.

Ismail, M. M., R. Samir, F. R. Saber, S. R. Ahmed, and M. A. Farag, "Oil as A Potential Treatment for Wound Infection: In Vitro and In Vivo Bioassays in Relation to Its Chemical Composition.", Antibiotics (Basel, Switzerland), vol. 9, issue 10, 2020. Abstract

Bacterial biofilm contributes to antibiotic resistance. Developing antibiofilm agents, more favored from natural origin, is a potential method for treatment of highly virulent multidrug resistant (MDR) bacterial strains; The potential of and essential oils (E.Os) antibacterial and antibiofilm activities in relation to their chemical composition, in addition to their ability to treat wound infection in mice model were investigated; leaf E.O at 0.05 µg·mL efficiently inhibited and eradicated biofilm formed by by 85% and 34%, respectively. Both and leaf E.Os showed a bactericidal action against within 6h at 2.08 µg·mL. In addition, a significant reduction of microbial load in mice wound infection model was found. Furthermore, gas chromatography mass spectrometry analysis revealed qualitative and quantitative differences among and leaf and berry E.Os. Monoterpene hydrocarbons, oxygenated monoterpenes, and phenolics were the major detected classes. β-Myrcene, limonene, 1,8-cineole, and eugenol were the most abundant volatiles. While, sesquiterpenes were found as minor components in berries E.O; Our finding suggests the potential antimicrobial activity of leaf E.O against MDR wound infections and their underlying mechanism and to be further tested clinically as treatment for MDR infections.

Fahmy, H., N. Hegazi, S. El-Shamy, and M. A. Farag, "Pomegranate juice as a functional food: a comprehensive review of its polyphenols, therapeutic merits, and recent patents.", Food & function, vol. 11, issue 7, pp. 5768-5781, 2020. Abstract

Pomegranate (Punica granatum) is an ancient perennial plant species of the Punicaceae family and is regarded as the 'miracle fruit' for its seeds being consumed as food, juice and as a functional food. Significant modern pharmacological and clinical evidence has highlighted the wide medicinal applications of pomegranate fruit parts and its juice. Pomegranate juice (PJ) that is superior to other fruit juices is a fortified source of dietary polyphenols with potential antioxidant capacity. Polyphenols of PJ include tannins, anthocyanins, and flavonoids. The presence of these beneficial phytochemicals is directly linked to its favourable health benefits viz., obesity and diabetes management and anti-inflammatory effects. This comprehensive review capitalizes on PJ with emphasis on the interrelationship between its holistic chemical composition, metabolism and biological effects. Moreover, the review recapitulates on the diverse health benefits of PJ and related patents in the field of PJ production to ensure the best produced juice quality.

Abdallah, H. M. I., N. M. Ammar, M. F. Abdelhameed, A. E. - N. E. G. Gendy, T. I. M. Ragab, A. M. Abd-ElGawad, M. A. Farag, M. S. Alwahibi, and A. I. Elshamy, "Protective Mechanism of Butanol Extract and Its Nano-Formulations against Ulcerative Colitis in Rats as Revealed via Biochemical and Metabolomic Assays.", Biology, vol. 9, issue 8, 2020. Abstract

Ulcerative colitis (UC) is a relapsing inflammatory disease of unknown etiology. The increased risk of cancer in UC patients warrants for the development of novel drug treatments. Herein, this work concerns with the investigation of the protective effects of butanol extract (ASBE) and its nanoformulations on UC in a rat model and its underlying mechanism. Colitis was induced by slow intrarectal infusion of 2 mL of 4% (v/v in 0.9% saline) acetic acid. Colon samples were evaluated macroscopically, microscopically, and assayed for pro-inflammatory cytokine levels. To monitor associated metabolic changes in acetic acid-induced UC model, serum samples were analyzed for primary metabolites using GC-MS followed by multivariate data analyses. Treatment with ASBE attenuated acetic acid-induced UC as revealed by reduction of colon weight, ulcer area, and ulcer index. ASBE treatment also reduced Cyclooxygenase-2 (COX-2), Prostaglandin E2 (PGE2) & Interleukin-1 (IL-1) levels in the inflamed colon. The nano-formulation of ASBE showed better protection than the crude extract against ulcer indices, increased PGE2 production, and histopathological alterations such as intestinal mucosal lesions and inflammatory infiltration. Distinct metabolite changes were recorded in colitis rats including a decrease in oleamide and arachidonic acid along with increased levels of lactic acid, fructose, and pyroglutamic acid. Treatment with nano extract restored metabolite levels to normal and suggests that cytokine levels were regulated by nano extract in UC. Conclusion: ASBE nano extract mitigated against acetic acid-induced colitis in rats, and the underlying mechanism could be attributed to the modulatory effects of ASBE on the inflammatory cascades. The applicability of metabolomics developed in this rat model seems to be crucial for evaluating the anti-inflammatory mechanisms of new therapeutics for acute colitis.