Feature-Based Molecular Networking for the Exploration of the Metabolome Diversity of Common Egyptian Centaurea Species in Relation to Their Cytotoxic Activity.

Centaurea is a genus compromising over 250 herbaceous flowering species and is used traditionally to treat several ailments. Among the Egyptian Centaurea species, C. lipii was reported to be cytotoxic against multidrug-resistant cancer cells. In this context, we aimed to explore the metabolome of C. lipii and compare it to other members of the genus in pursuance of identifying its bioactive principles. An LC-MS/MS analysis approach synchronized with feature-based molecular networks was adopted to offer a holistic overview of the metabolome diversity of the Egyptian Centaurea species. The studied plants included C. alexandrina, C. calcitrapa, C. eryngioides, C. glomerata, C. lipii, C. pallescens, C. pumilio, and C. scoparia. Their constitutive metabolome showed diverse chemical classes such as cinnamic acids, sesquiterpene lactones, flavonoids, and lignans. Linking the recorded metabolome to the previously reported cytotoxicity identified sesquiterpene lactones as the major contributors to this activity. To confirm our findings, bioassay-guided fractionation of C. lipii was adopted and led to the isolation of the sesquiterpene lactone cynaropicrin with an IC50 of 1.817 µM against the CCRF-CEM leukemia cell line. The adopted methodology highlighted the uniqueness of the constitutive metabolome of C. lipii and determined the sesquiterpene lactones to be the responsible cytotoxic metabolites.

Plant cell cultures: An enzymatic tool for polyphenolic and flavonoid transformations.

BACKGROUND: In the pharmaceutical sector, tissue culture techniques for large-scale production of natural chemicals can be a less expensive alternative to large-scale synthesis. Although recent biotransformation research have used plant cell cultures to target a wide range of bioactive compounds, more compiled information and synopses are needed to better understand metabolic pathways and improve biotransformation efficiencies. PURPOSE: This report reviews the biochemical transformation of phenolic natural products by plant cell cultures in order to identify potential novel biotechnological approaches for ensuring more homogeneous and stable phenolic production year-round under controlled environmental conditions. METHODS: Articles on the use of plant cell culture for polyphenolic and flavonoid transformations (1988 - 2021) were retrieved from SciFinder, PubMed, Scopus, and Web of Science through electronic and manual search in English. Following that, the authors chose the required papers based on the criteria they defined. The following keywords were used for the online search: biotransformation, Plant cell cultures, flavonoids, phenolics, and pharmaceutical products. RESULTS: The initial search found a total of 96 articles. However, only 70 of them were selected as they met the inclusion criteria defined by the authors. The analysis of these studies revealed that plant tissue culture is applicable for the large-scale production of plant secondary metabolites including the phenolics, which have high therapeutic value. CONCLUSION: Plant tissue cultures could be employed as an efficient technique for producing secondary metabolites including phenolics. Phenolics possess a wide range of therapeutic benefits, as anti-oxidant, anti-cancer, and anti-inflammatory properties. Callus culture, suspension cultures, transformation, and other procedures have been used to improve the synthesis of phenolics. Their production on a large scale is now achievable. More breakthroughs will lead to newer insights and, without a doubt, to a new era of phenolics-based pharmacological agents for the treatment of a variety of infectious and degenerative disorders.

Comparative Study on the Essential Oils from Five Wild Egyptian Centaurea Species: Effective Extraction Techniques, Antimicrobial Activity and In-Silico Analyses.

The genus Centaurea is recognized in folk medicine for anti-inflammatory, anti-itch, antitussive, purgative, astringent, and tonic activities. To study the chemical determinant for antimicrobial activity essential oils (EOs), five Centaurea species were analyzed including: C. scoparia, C. calcitrapa, C. glomerata, C. lipii and C. alexandrina. Conventional hydro-distillation (HD) and microwave-assisted extraction (MAE), as new green technologies, were compared for the extraction of essential oils. GC/MS analysis identified 120 EOs including mostly terpenoid except from C. lipii and C. alexandrina in which nonterpenoids were the major constituents. Major terpenoids included spathulenol, caryophyllene oxide and alloaromadendrene oxide-2. To probe antibacterial activity, potential EO inhibitors of a bacterial type II DNA topoisomerase, DNA gyrase B were screened via an in silico molecular docking approach. Spathulenol and alloaromadendrene oxide-2 possessed the best binding affinity in the ATP- binding pocket of Gyrase B enzyme. Principal component analysis and agglomerative hierarchical clustering were used for sample classification and revealed that sesquiterpenes contributed the most for accessions classification. In vitro antimicrobial activity against Staphylococcus aureus, Escherichia coli and Aspergillus niger for all EOs were also evaluated. EOs from C. lipii, C. glomerata and C. calcitrapa exhibited significant MIC against S. aureus with an MIC value of 31.25 µg/mL.

New antimicrobial metabolites from the medicinal herb Artemisia herba-Alba.

Artemisia herba-alba is widely used in traditional medicines for the treatment of several diseases. From the aerial parts organic extract of A. herba-alba, two new compounds, 1,3,8-trihydroxyeudesm-4-en-7α,11ßH-12,6α-olide (1) and 5-ß-​D-​glucopyranosyloxy​-​7-methoxy-​6H-​benzopyran-​2-​one (2), respectively, together with five known metabolites: 3α,8ß-dihydroxygermacr-4(15),9(10)-dien-7ß,11αH,12,6α-olide (3), 1ß,8α-dihydroxy-11α,13-dihydrobalchanin (4), 11-epiartapshin (5), tomenin (6) and benzoic acid, p-​(ß-​D-​glucopyranosyloxy)​-​methyl ester (7), were isolated and identified. The chemical structures were proven depending upon spectroscopic analysis, including 1 D/2D NMR as and ESI-MS. Compound 1 inhibited Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus. Compounds 2 and 3 exhibited antibacterial activity against both gram-positive and gram-negative bacteria.

Prevention of Infective Endocarditis of Oral Origin in Children: A Call for Pediatric Dentists-Pediatric Cardiologists' Collaboration in Egypt.

AIM: To assess knowledge and practices of some Egyptian pediatric dentists and pediatric cardiologists/cardiac surgeons regarding prevention of infective endocarditis (IE) from oral origin in children. MATERIALS AND METHODS: Pediatric dentists or pediatric cardiologists/cardiac surgeons having their practice in Egypt were conveniently selected. An online questionnaire was constructed for each specialty practitioner in English using a web-based platform. Each survey tool collected data about demographics, knowledge, and practices concerning the prevention of IE of oral origin. Surveys were disseminated to potential respondents using direct messaging or posting surveys' links on high-traffic areas (blogs, groups, or pages) of commonly known social media platforms. RESULTS: Two-hundred and thirty-nine pediatric dentists and 71 pediatric cardiologists/cardiac surgeons' responses were included in analysis. Some shortage in oral health knowledge and deviations in management protocol from known guidelines were evident in cardiologists' responses, whereas 66.2 and 64.8% reported encountering IE of oral origin or canceling a cardiac surgery due to oral infection, respectively. Most pediatric dentists (65.7%) rely on physician referral before managing children at risk of IE despite following recognized guidelines for IE prevention which may reflect difficulty in understanding such guidelines. Ninety percent of children with heart diseases visit a dentist due to oral complaint and not for checkup. CONCLUSION: Identification of disparities and pitfalls in management of children with heart diseases, if appropriately addressed by pediatric dentists and cardiologists, may reduce the risk of IE from an oral origin. CLINICAL SIGNIFICANCE: Health education of both specialists and development of national guidelines based on national epidemiology and clinical experience of cardiologists are strongly needed to decrease oral disease burden, allow for a consensus of patient management, and minimize the need for cross-referral, thus facilitating dental management without undue delays.

When the left atrium becomes a monster: a case report.

BACKGROUND: Congenital left atrium (LA) aneurysms are extremely rare entities in clinical practice and most frequently involve the atrial appendage and rarely arise from the body of LA, We report a case of giant LA aneurysm compressing heart and presenting in a very late stage. CASE SUMMARY: A 31-year-old male, who was diagnosed to have dextrocardia, rheumatic heart disease, and atrial fibrillation and was kept on medical treatment long time ago, presented with congestive heart failure symptoms and cardiogenic shock. Emergency transthoracic echocardiography was done revealing situs solitus with aneurysmally dilated LA pushing heart to the right side (dextro-posed heart), moderate mitral regurgitation, and severe pulmonary hypertension, however, pulmonary artery anatomy could not be properly visualized so computed tomography (CT) was preformed confirming diagnosis and revealing compressed pulmonary arterial tree by the dilated LA, unfortunately patient died before proceeding to surgical intervention. DISCUSSION: Congenital left atrial aneurysms are extremely rare anomaly and may be associated with significant morbidity. And, therefore, should be remembered as a potential anatomic cause of atrial arrhythmias or embolic phenomena, or both. The diagnosis may be easily established through non-invasive complementary techniques, such as echocardiography, CT, and cardiac magnetic resonance imaging. Symptomatic patients, those with large aneurysm or compelling indications for surgery should undergo surgical resection.

Terpenoid bio-transformations and applications via cell/organ cultures: a systematic review.

Structurally diverse natural products are valued for their targeted biological activity. The challenge of working with such metabolites is their low natural abundance and complex structure, often with multiple stereocenters, precludes large-scale or unsophisticated chemical synthesis. Since select plants contain the enzymatic machinery necessary to produce specialized compounds, tissue cultures can be used to achieve key transformations for large-scale chemical and/or pharmaceutical applications. In this context, plant tissue-culture bio-transformations have demonstrated great promise in the preparation of pharmaceutical products. This review describes the capacity of cultured plant cells to transform terpenoid natural products and the specific application of such transformations over the past three decades (1988-2019).

Cytotoxicity of 40 Egyptian plant extracts targeting mechanisms of drug-resistant cancer cells.

BACKGROUND: The multidrug resistance (MDR) phenotype encounters a major challenge to the success of established chemotherapy in cancer patients. We hypothesized that cytotoxic medicinal plants with novel phytochemicals can overcome MDR and kill MDR-cells with similar efficacy as drug sensitive cells. PURPOSE: We evaluated plant extracts from an unexplored ecosystem in Egypt with unusual climate and nutrient conditions for their activity against sensitive and multidrug-resistant cancer cell lines. MATERIAL AND METHODS/STUDY DESIGN: Methylene chloride: methanol (1:1) and methanol: H2O (7:3) extracts of 40 plants were prepared resulting in a sum of 76 fraction containing compounds with varying polarity. The resazurin reduction assay was employed to evaluate the cytotoxicity of these extracts on five matched pairs of drug-sensitive and their drug-resistant cell lines. Flow cytometry and Western blotting was used to determine cell cycle analyses, apoptosis, and autophagy. Reactive oxygen species (ROS) were measured spectrophotometrically. RESULTS: Extracts derived from Withania obtusifolia (WO), Jasonia candicans (JC), Centaurea lippii (CL), and Pulicaria undulata (PU) were the most active ones among 76 extracts from 40 Egyptian medicinal plants. They showed a significant reduction of cell viability on drug-sensitive CCRF-CEM leukemia cell line with IC50 values less than 7 µg/ml. Low cross-resistance degrees were observed in multidrug-resistant CEM/ADR5000 cells towards CL (1.82-fold) and JC (6.09-fold). All other drug-resistant cell lines did not reveal cross-resistance to the four extracts. Further mechanistic assessment have been studied for these four extracts. CONCLUSION: The methylene chloride: methanol (1:1) fractions of WO, JC, CL, and PU are promising cytotoxic extracts that could be used to combat MDR cancer cells through different cell death pathways.

Immediate and short-term effect of balloon mitral valvuloplasty on global and regional biventricular function: a two-dimensional strain echocardiographic study.

AIM: To assess the effect of balloon mitral valvuloplasty (BMV) on global and regional ventricular functions using 2D strain. METHODS AND RESULTS: Thirty-two patients with mitral stenosis (MS) and 30 healthy subjects underwent full echocardiographic examinations, including left ventricle (LV) and right ventricle (RV) regional and global longitudinal strain (GLS) measurements. In MS patients, measurements were repeated within 24 h and 3 months after BMV. Patients with MS had lower LV and RV GLS compared with control group (-16.5 ± 2.7% vs. -21.0 ± 1.5) and (-18.3 ± 4.7 vs. -19.8 ± 1.3), respectively. Significant decrease was noted in the basal and septal segments compared with the apical LV segments and RV free wall. BMV resulted in significant improvement in LV and RV GLS within 24 h post-BMV compared with baseline values (P = 0.0001 and 0.0002, respectively), an improvement which was maintained after 3 months. There was significant positive correlation between both LV and RV GLS at baseline and mitral valve mean pressure gradient and RV systolic pressure and significant inverse correlation between LV GLS and MVA. CONCLUSION: MS patients have subclinical LV and RV systolic dysfunction by GLS despite normal ejection fraction and fractional area change. BMV results in marked improvement in LV and RV GLS immediately post-BMV with trend towards normalization at follow-up after 3 months. A mixed aetiology theory involving a myocardial as well as a haemodynamic factor is believed to be the cause for this subclinical biventricular dysfunction and its improvement at short-term follow-up post-BMV.

Evaluation of the essential oil of Foeniculum vulgare Mill (fennel) fruits extracted by three different extraction methods by GC/MS.

BACKGROUND: Hydrodistillation (HD) and steam-distillation, or solvent extraction methods of essential oils have some disadvantages like thermal decomposition of extracts, its contamination with solvent or solvent residues and the pollution of residual vegetal material with solvent which can be also an environmental problem. Thus, new green techniques, such as supercritical fluid extraction and microwave assisted techniques, are potential solutions to overcome these disadvantages. MATERIALS AND METHODS: The aim of this study was to evaluate the essential oil of Foeniculum vulgare subsp. Piperitum fruits extracted by three different extraction methods viz. Supercritical fluid extraction (SFE) using CO2, microwave-assisted extraction (MAE) and hydro-distillation (HD) using gas chromatography-mass spectrometry (GC/MS). RESULTS: The results revealed that both MAE and SFE enhanced the extraction efficiency of the interested components. MAE gave the highest yield of oil as well as higher percentage of Fenchone (28%), whereas SFE gave the highest percentage of anethol (72%). CONCLUSION: Microwave-assisted extraction (MAE) and supercritical fluid extraction (SFE) not only enhanced the essential oil extraction but also saved time, reduced the solvents use and produced, ecologically, green technologies.

Lipophilic Constituents of Rumex vesicarius L. and Rumex dentatus L.

Rumex dentatus L. and Rumex vesicarius L., of the family Polygonaceae, are edible herbs growing wild in Egypt. Their lipoid constituents were examined by both liquid chromatography/mass spectrometry (LC/MS) and by gas chromatography/mass spectrometry (GC/MS). Their essential oil compositions consisted mainly of thujene, limonene, fenchon, estragole, and anethole but at largely different concentration. Fatty acid compositions were similar among the two species and consisting of palmitic, oleic, linoleic and linolenic acids, with R. vesicarius containing much higher level of omega-3-fatty acids. Both of the crude lipid extracts of the two species showed strong antioxidant activity as a radical quenching agent against 2,2-diphenyl-1-picrylhydrazyl (DPPH) systems. Antioxidant activities were mostly associated with the polar lipid fractions. High performance thin layer chromatography (HPTLC), both in the normal and reversed phase,as well as liquid chromatography/mass spectrometry (LC/MS) in the positive and negative electrospray ionization (ESI), showed unique chemical profile for each species that can be useful for species identification and quality control of herbal drug formulations. R. vesicarius was characterized by abundances of flavonoids and R. dentatus was abundant in anthraquinones and chromones.

Isolation and characterization of antineoplastic alkaloids from Catharanthus roseus L. Don. cultivated in Egypt.

Vinblastine and vincristine (the antileukemic agents) were isolated, in a pure form, from Catharanthus roseus L. Don., cultivated in Egypt, by several chromatographic techniques. Five modified methods for the preparation of total alkaloids were carried out. All the isolated mixtures were evaluated by HPLC and HPTLC analyses. The antineoplastic alkaloids; vinblastine and vincristine, were isolated by the use of vacuum liquid chromatographic column on silica gel : aluminium oxide (1:1) mixed bed vacuum liquid chromatography (VLC), Charcoal column, and finally purified by centrifugally accelerated radial chromatography (Chromatotrone).

Lipid fraction constituents and evaluation of anti-anaphylactic activity of Prunus mahaleb L. kernels.

The lipid fraction constituents as well as evaluation of anti-anaphylactic activity of Prunus mahaleb L. kernels were studied. Prunus mahaleb L. kernels were obtained from the local market in Cairo, Egypt. Investigation of the fatty acids revealed that oleic and linoleic acids are the major constituents. 12 compounds were identified from the hydrocarbon fraction. The sterol fraction comprises of cholesterol, stigmasterol, beta-sitosterol and campesterol. The pharmacotoxicity studies were carried out on total and defatted ethanolic extracts as well as the oil fraction. The oil fraction proved to be extremely safe and free from any acute lethal toxicity in intraperitoneal (i.p.) and oral doses up to 100 ml/kg. In vivo assessment of prophylactic efficacy was afforded by 7 days course of daily medication schedule of sensitized adult male guinea pigs against ovalbumin bronchospasm. The prophylactic anti-inflammatory activity of the total ethanolic extract was higher than that of the defatted ethanolic extract. In addition, the lipid fraction of Prunus mahaleb L. kernels evoked complete anti-inflammatory efficacy among the survival animals receiving low and medium doses.

Naphthaquinones of Alkanna orientalis (L.) Boiss.

The roots of Alkanna orientalis (L.) Boiss yielded alpha-methyl-n-butyl alkannin (compound 1) and alkannin acetate (compound 2). The compounds were identified by UV, MS, (1)H NMR and (13)C NMR. Quantitative determination of alpha-methyl-n-butyl alkannin and alkannin acetate in Alkanna orientalis (L) Boiss roots was established by TLC densitometry.