Quantitative Analysis of Isopimpinellin from Ammi majus L. Fruits and Evaluation of Its Biological Effect on Selected Human Tumor Cells.

Ammi majus L. (Apiaceae) is a medicinal plant with a well-documented history in phytotherapy. The aim of the present work was to isolate isopimpinellin (5,8-methoxypsoralen; IsoP) from the fruit of this plant and evaluate its biological activity against selected tumor cell lines. The methanol extract obtained with the use of an accelerated solvent extraction (ASE) method was the most suitable for the quantitative analysis of coumarins in the A. majus fruit matrix. The coumarin content was estimated by RP-HPLC/DAD, and the amount of IsoP was found to be 404.14 mg/100 g dry wt., constituting 24.56% of the total coumarin fraction (1.65 g/100 g). This, along with the presence of xanthotoxin (368.04 mg/100 g, 22.36%) and bergapten (253.05 mg/100 g, 15.38%), confirmed A. majus fruits as an excellent source of these compounds. IsoP was isolated (99.8% purity) by combined liquid chromatography/centrifugal partition chromatography (LC/CPC) and tested for the first time on its antiproliferative activity against human colorectal adenocarcinoma (HT29, SW620), osteosarcoma (Saos-2, HOS), and multiple myeloma (RPMI8226, U266) cell lines. MTT assay results (96 h incubation) demonstrated a dose- and cell line-dependent decrease in cell proliferation/viability, with the strongest effect of IsoP against the Saos-2 cell line (IC50; 42.59 µM), medium effect against U266, HT-29, and RPMI8226 (IC50 = 84.14, 95.53, and 105.0 µM, respectively), and very weak activity against invasive HOS (IC50; 321.6 µM) and SW620 (IC50; 711.30 µM) cells, as well as normal human skin fibroblasts (HSFs), with IC50; 410.7 µM. The mechanistic study on the Saos-2 cell line showed that IsoP was able to reduce DNA synthesis and trigger apoptosis via caspase-3 activation. In general, IsoP was found to have more potency towards cancerous cells (except for HOS and SW620) than against healthy cells. The Selective Index (SI) was determined, underlining the higher selectivity of IsoP towards cancer cells compared to healthy cells (SI = 9.62 against Saos-2). All these results suggest that IsoP might be a promising molecule in the chemo-prevention and treatment of primary osteosarcoma.

Antibacterial effects of chitosan-based hydrogels containing Trachyspermum ammi essential oil on pathogens isolated from dogs with otitis externa.

BACKGROUND: Growing antibiotic resistance has made treating otitis externa (OE) increasingly challenging. On the other hand, local antimicrobial treatments, especially those that combine essential oils (EOs) with nanoparticles, tend to be preferred over systemic ones. It was investigated whether Ajwain (Trachyspermum ammi) EO, combined with chitosan nanoparticles modified by cholesterol, could inhibit the growth of bacterial pathogens isolated from OE cases in dogs. In total, 57 dogs with clinical signs of OE were examined and bacteriologically tested. Hydrogels of Chitosan were synthesized by self-assembly and investigated. EO was extracted (Clevenger machine), and its ingredients were checked (GC-MS analysis) and encapsulated in chitosan-cholesterol nanoparticles. Disc-diffusion and broth Micro-dilution (MIC and MBC) examined its antimicrobial and therapeutic properties. RESULTS: Staphylococcus pseudintermedius (49.3%) was the most common bacteria isolated from OE cases, followed by Pseudomonas aeruginosa (14.7%), Escherichia coli (13.3%), Streptococcus canis (9.3%), Corynebacterium auriscanis (6.7%), Klebsiella pneumoniae (2.7%), Proteus mirabilis (2.7%), and Bacillus cereus (1.3%). The investigation into the antimicrobial properties of Ajwain EO encapsulated in chitosan nanoparticles revealed that it exhibited a more pronounced antimicrobial effect against the pathogens responsible for OE. CONCLUSIONS: Using chitosan nanoparticles encapsulated with EO presents an effective treatment approach for dogs with OE that conventional antimicrobial treatments have not cured. This approach not only enhances antibacterial effects but also reduces the required dosage of antimicrobials, potentially preventing the emergence of antimicrobial resistance.

Assessment of Trachyspermum ammi essential oil against Aspergillus flavus, aflatoxin B1 contamination, and post-harvest quality of Sorghum bicolor.

The present investigation explored the antifungal effectiveness of Trachyspermum ammi essential oil (TAEO) against Aspergillus flavus, aflatoxin B1 (AFB1) contamination, and its mechanism of action using biochemical and computational approaches. The GC-MS result revealed the chemical diversity of TAEO with the highest percentage of γ-terpinene (39 %). The TAEO exhibited minimum inhibitory concentration against A. flavus growth (0.5 µL/mL) and AFB1 (0.4 µL/mL) with radical scavenging activity (IC50 = 2.13 µL/mL). The mechanism of action of TAEO was associated with the alteration in plasma membrane functioning, antioxidative defense, and carbon source catabolism. The molecular dynamic result shows the multi-regime binding of γ-terpinene with the target proteins (Nor1, Omt1, and Vbs) of AFB1 biosynthesis. Furthermore, TAEO exhibited remarkable in-situ protection of Sorghum bicolor seed samples against A. flavus and AFB1 contamination and protected the nutritional deterioration. Hence, the study recommends TAEO as a natural antifungal agent for food protection against A. flavus mediated biodeterioration.

Exploring the multifaceted effects of Ammi visnaga: subchronic toxicity, antioxidant capacity, immunomodulatory, and anti-inflammatory activities.

Ammi visnaga (A. visnaga) is an annual herb that has been used in traditional medicine to treat various ailments attributed to the presence of its bioactive compounds. The purpose of this study was to identify and examine the phytochemical properties of the hydroalcoholic extract of A. visnaga using in vitro and in vivo models. Our findings demonstrated that the extract contained a variety of beneficial components, including phenols, flavonoids, tannins, coumarins, saponins, khellin, and visnagin. The total polyphenolic content and total flavonoid content were 23.26 mg/GAE/g dry weight and 13.26 mg/GAE/g dry weight, respectively. In vitro tests demonstrated that the extract possessed antioxidant properties as evidenced by the ability to scavenge free radicals, including DPPH, ABTS, nitric oxide (NO), phosphomolybdate, and ferric-reducing antioxidant power (FRAP). Further, the extract was found to inhibit hydrogen peroxide (H2O2)-induced hemolysis. In a 90-d in vivo study, female Wistar rats were administered 1 g/kg of A. visnaga extract orally resulting in a significant increase in total white blood cell count. Although morphological changes were observed in the liver, no marked alterations were noted in kidneys and spleen. In a female Swiss albino mice model of acetic acid-induced vascular permeability, A. visnaga significantly inhibited extravasations of Evans blue at doses of 0.5 or 1 g/kg with inhibition percentages of 51 and 65%, respectively, blocking tissue necrosis. The extract also demonstrated potential immunomodulatory properties in mice by enhancing antibody production in response to antigens. In silico molecular docking studies demonstrated a strong affinity between khellin or visnagin and immunomodulatory proteins, NF-κB, p52, and TNF-α. These findings suggest that A. visnaga may be considered a beneficial antioxidant with immunomodulatory properties and might serve as a therapeutic agent to combat certain diseases.

Nanoliposomal Trachyspermum ammi (L) sprague essential oil for effective control of malaria mosquito larvae, Anopheles stephensi Liston.

Controlling mosquito vectors at immature stages using larvicides is a practical strategy to stave off mosquito-borne diseases such as malaria. Developing nanoliposomes bearing essential oil is a promising approach to improving the efficacy and stability of EOs-derived larvicides. The main aim of this investigation was to assess the efficacy of nanoliposome containing Trachyspermum ammi L. EO (TAEO-NL) as a new potential formulation to control Anopheles stephensi Liston (Diptera, Culicidae) mosquito larvae. The chemical constituents of T. ammi L. essential oil (TAEO) were first investigated using gas chromatography-mass spectrometry (GC-MS) analysis; its dominant component (48.22%) was thymol. TAEO-NL with a particle size of 54.6 ± 5 nm and zeta potential of -18 ± 0.5 mV were then prepared using the ethanol injection method. Besides, the successful loading of TAEO was confirmed using Attenuated Total Reflection-Fourier Transform Infra-Red (ATR-FTIR) spectroscopy analysis. A significant difference (P < 0.05) was observed in the efficacy of TAEO-NL and TAEO with lethal concentration 50% (LC50) values of 14.09 and 59.47 µg/mL against An. stephensi larvae. However, free nanoliposomes show negligible larvicidal effects (<5%). This nano-formulation could thus be suggested as a green product against insects to impede transmission of deadly infectious diseases with possible field applicability scope.

Molecular Docking and Dynamics Simulations of Ammi visnaga L. Constituents as Antimelanogenic, Anti-Inflammatory and Anticoagulant Agents.

In this study, anti-melanogenic, anti-inflammatory and anti-coagulant potentials of eighteen selected constituents of Ammi visnaga L. were investigated by Induced Fit Docking (IFD) and molecular dynamic simulation with Schrödinger software. The binding free energies of the selected natural compounds were computed by means of ΔG MM-GBSA studies. Anti-melanogetic activity of the constituent against agaricus bisporus tyrosinase, Priestia megaterium tyrosinase and Homo sapiens tyrosinase were evaluated. The result showed that apiumetin had more negative binding free energy against three tyrosinase enzymes than cognate ligands, tropolone and kojic acid. Docking analysis was also performed to predict the constituents with anti-inflammatory activity against human Tumor necrosis factor, Cyclooxygenase-2, Prostaglandin D2 11-ketoreductase AKR1C3 and Prostaglandin reductase PTGR2. The results showed that pyranocoumarins (visnadin, dihydrosamidin, samidin) have more negative binding free energy against Cyclooxygenase-2 and Prostaglandin D2 11-ketoreductase receptors than cognate drugs, rofecoxib and indomethacin. In addition, docking analysis shows that pyranocoumarins, apiumetin and cimifugin have more negative binding free energy against Vitamin K epoxide reductase than S-warfarin drug, predicting that they have anticoagulant activity. Furthermore, the constituents and their cognate drugs were subjected to 100 ns MD Simulation to predict their stability at the active sites of the enzymes.

Biochemical and phytochemical responses of Ammi visnaga L. (Apiaceae) callus culture elicited by SiO2 and graphene Oxide-SiO2 nanoparticles.

Ammi visnaga L. is an enriched medicinal plant with medicinally important compounds. Two types of nanoparticles (NPs) including silica (SiO2) and graphene oxide bound with SiO2 (GO-SiO2) NPs at different concentrations (0, 15, 25 mg L-1) were used as elicitors to investigate their effects on callus morphology, H2O2 content, total phenolic content (TPC), total flavonoids content (TFC), ferric reducing/antioxidant power (FRAP), and few antioxidant enzymes such as catalase (CAT), guaiacol peroxidase (GPX), superoxide dismutase (SOD), ascorbate peroxidase (APX), and polyphenol oxidase (PPO) in the callus cultures of A. visnaga. The effects of elicitation of both NPs on calli were observed using a scanning electron microscope (SEM). The 15 mg L-1 concentration of GO-SiO2 NPs produced the highest TPC (193.3 mg GAE g-1 FW), CAT (13.1 U mg-1 Protein), GPX (0.0089 U mg-1 Protein), and APX (0.079 U mg-1 Protein). Whereas, the maximum content of H2O2 (0.68 µmol g-1 FW), FRAP (0.0092 µmol mg-1), and TFC (62.27 mg QE g-1 FW) was observed at 25 mg L-1 and 15 mg L-1 of SiO2 NPs, respectively. Conclusively, in the callus culture of A. visnaga, the 15 mg L-1 concentration of GO-SiO2 NPs was the most suitable dosage for enhancing the enzymatic antioxidant activities (CAT, GPX, APX) and TPC, rather than SiO2 NPs.

Thymol in Trachyspermum ammi seed extract exhibits neuroprotection, learning, and memory enhancement in scopolamine-induced Alzheimer's disease mouse model.

Several reports have stated the neuroprotective and learning/memory effects of Tachyspermum ammi seed extract (TASE) and its principal component thymol; however, little is known about its underlying molecular mechanisms and neurogenesis potential. This study aimed to provide insights into TASE and a thymol-mediated multifactorial therapeutic approach in a scopolamine-induced Alzheimer's disease (AD) mouse model. TASE and thymol supplementation significantly reduced oxidative stress markers such as brain glutathione, hydrogen peroxide, and malondialdehyde in mouse whole brain homogenates. Tumor necrosis factor-alpha was significantly downregulated, whereas the elevation of brain-derived neurotrophic factor and phospho-glycogen synthase kinase-3 beta (serine 9) enhanced learning and memory in the TASE- and thymol-treated groups. A significant reduction in the accumulation of Aß 1-42 peptides was observed in the brains of TASE- and thymol-treated mice. Furthermore, TASE and thymol significantly promoted adult neurogenesis, with increased doublecortin positive neurons in the subgranular and polymorphic zones of the dentate gyrus in treated-mice. Collectively, TASE and thymol could  potentially act as natural therapeutic agents for the treatment of  neurodegenerative disorders, such as  AD.

Membrane reactor for production of biodiesel from nonedible seed oil of Trachyspermum ammi using heterogenous green nanocatalyst of manganese oxide.

Conventional homogeneous-based catalyzed transesterification for the production of biodiesel can be replaced with a membrane reactor that has an immobilized heterogeneous catalyst. Combining reaction with separation while utilizing membranes with a certain pore size might boost conversion process. this investigation to study the effectiveness of membrane reactor in combination with heterogeneous green nano catalysis of MnO2. Techniques such as XRD, EDX, FTIR, SEM, and TGA were used to characterize the synthesized MnO2 nano catalyst. The highest conversion of around 94% Trachyspermum ammi oil was obtained by MnO2. The optimum process variables for maximum conversion were catalyst loading of 0.26 (wt.%), 8:1 M ratio, 90 °C reaction temperature, and time 120 min. The green nano catalyst of MnO2 was reusable up to five cycles with minimum loss in conversion rate of about 75% in the fifth cycle. Nuclear magnetic resonance validated the synthesis of methyl esters. It was concluded that membrane reactor a promising technique to efficiently transesterify triglycerides into methyl esters and enable process intensification uses MnO2 as a catalyst.

An Insight into Biochemical Characterization and Explorations of Antioxidant, Antibacterial, Cytotoxic, and Antidiabetic Activities by Trachyspermum ammi Nanosuspensions.

BACKGROUND: Trachyspermum ammi is a frequently utilized traditional medicinal plant renowned for its pharmacological attributes, particularly in the realm of treating infectious diseases. This current study aims to comprehensively assess the in vitro properties of freshly prepared nanosuspensions derived from Trachyspermum ammi extracts, with a focus on their cost-effective potential in various areas, including antioxidant, antibacterial, cytotoxic, and antidiabetic activities. METHODS: Biochemical characterization of T. ammi nanosuspensions by high-performance liquid chromatography (HPLC) and Fourier-transform infrared spectroscopy (FTIR) analyses. RESULTS: HPLC analysis revealed the presence of kaempferol and sinapic acid in various amounts at 11.5 ppm and 12.3 ppm, respectively. FTIR analysis of T. ammi powder revealed the presence of alcohols and amines. The assessment of antioxidant activity was conducted using a DPPH scavenging assay, indicating that the nanosuspensions exhibited their highest free radical scavenging activity, reaching 14.9%. Nanosuspensions showed 3.75 ± 3.529.5% biofilm inhibition activity against Escherichia coli. The antidiabetic activity was accessed through antiglycation and α- amylase inhibition assays, while nanosuspension showed the maximum inhibition activity at 25.35 ± 0.912133% and 34.6 ± 1.3675%. Hemolytic activity was also evaluated, and T. ammi nanosuspension showed 22.73 ± 1.539% hemolysis. CONCLUSIONS: This nanotechnology approach has established a foundation to produce plant-based nanosuspensions, offering a promising avenue for the biopharmaceutical production of herbal nanomedicines. These nanosuspensions have the potential to enhance bioavailability and can serve as a viable alternative to synthetic formulations.

Ultrasound-assisted nanoemulsion of Trachyspermum ammi essential oil and its constituent thymol on toxicity and biochemical aspect of Aedes aegypti.

Aedes aegypti is the main vector of yellow fever, chikungunya, Zika, and dengue worldwide and is managed by using chemical insecticides. Though effective, their indiscriminate use brings in associated problems on safety to non-target and the environment. This supports the use of plant-based essential oil (EO) formulations as they are safe to use with limited effect on non-target organisms. Quick volatility and degradation of EO are a hurdle in its use; the present study attempts to develop nanoemulsions (NE) of Trachyspermum ammi EO and its constituent thymol using Tween 80 as surfactant by ultrasonication method. The NE of EO had droplet size ranging from 65 ± 0.7 to 83 ± 0.09 nm and a poly dispersity index (PDI) value of 0.18 ± 0.003 to 0.20 ± 0.07 from 1 to 60 days of storage. The NE of thymol showed a droplet size ranging from 167 ± 1 to 230 ± 1 nm and PDI value of 0.30 ± 0.03 to 0.40 ± 0.008 from 1 to 60 days of storage. The droplet shape of both NEs appeared spherical under a transmission electron microscope (TEM). The larvicidal effect of NEs of EO and thymol was better than BEs (Bulk emulsion) of EO and thymol against Ae. aegypti. Among the NEs, thymol (LC50 34.89 ppm) had better larvicidal action than EO (LC50 46.73 ppm). Exposure to NEs of EO and thymol causes the shrinkage of the larval cuticle and inhibited the acetylcholinesterase (AChE) activity in Ae. aegypti. Our findings show the enhanced effect of NEs over BEs which facilitate its use as an alternative control measure for Ae. aegypti.

Plants Used in Mexican Traditional Medicine for the Management of Urolithiasis: A Review of Preclinical Evidence, Bioactive Compounds, and Molecular Mechanisms.

Urolithiasis (UL) involves the formation of stones in different parts of the urinary tract. UL is a health problem, and its prevalence has increased considerably in developing countries. Several regions use plants in traditional medicine as an alternative in the treatment or prevention of UL. Mexico has known about the role of traditional medicine in the management of urinary stones. Mexican traditional medicine uses plants such as Argemone mexicana L., Berberis trifoliata Hartw. ex Lindl., Costus mexicanus Liebm, Chenopodium album L., Ammi visnaga (L.) Lam., Eysenhardtia polystachya (Ortega) Sarg., Selaginella lepidophylla (Hook. & Grev.) Spring, and Taraxacum officinale L. These plants contain different bioactive compounds, including polyphenols, flavonoids, phytosterols, saponins, furanochromones, alkaloids, and terpenoids, which could be effective in preventing the process of stone formation. Evidence suggests that their beneficial effects might be associated with litholytic, antispasmodic, and diuretic activities, as well as an inhibitory effect on crystallization, nucleation, and aggregation of crystals. The molecular mechanisms involving these effects could be related to antioxidant, anti-inflammatory, and antimicrobial properties. Thus, the review aims to summarize the preclinical evidence, bioactive compounds, and molecular mechanisms of the plants used in Mexican traditional medicine for the management of UL.

Potential Anticancer Activity of the Furanocoumarin Derivative Xanthotoxin Isolated from Ammi majus L. Fruits: In Vitro and In Silico Studies.

Ammi majus L., an indigenous plant in Egypt, is widely used in traditional medicine due to its various pharmacological properties. We aimed to evaluate the anticancer properties of Ammi majus fruit methanol extract (AME) against liver cancer and to elucidate the active compound(s) and their mechanisms of action. Three fractions from AME (Hexane, CH2Cl2, and EtOAc) were tested for their anticancer activities against HepG2 cell line in vitro (cytotoxicity assay, cell cycle analysis, annexin V-FITC apoptosis assay, and autophagy efflux assay) and in silico (molecular docking). Among the AME fractions, CH2Cl2 fraction revealed the most potent cytotoxic activity. The structures of compounds isolated from the CH2Cl2 fraction were elucidated using 1H- and 13C-NMR and found that Compound 1 (xanthotoxin) has the strongest cytotoxic activity against HepG2 cells (IC50 6.9 ± 1.07 µg/mL). Treating HepG2 cells with 6.9 µg/mL of xanthotoxin induced significant changes in the DNA-cell cycle (increases in apoptotic pre-G1 and G2/M phases and a decrease in the S-phase). Xanthotoxin induced significant increase in Annexin-V-positive HepG2 cells both at the early and late stages of apoptosis, as well as a significant decrease in autophagic flux in cancer compared with control cells. In silico analysis of xanthotoxin against the DNA-relaxing enzyme topoisomease II (PDB code: 3QX3) revealed strong interaction with the key amino acid Asp479 in a similar fashion to that of the co-crystallized inhibitor (etoposide), implying that xanthotoxin has a potential of a broad-spectrum anticancer activity. Our results indicate that xanthotoxin exhibits anticancer effects with good biocompatibility toward normal human cells. Further studies are needed to optimize its antitumor efficacy, toxicity, solubility, and pharmacokinetics.

Macroporous resin-assisted enrichment, characterizations, antioxidant and anticandidal potential of phytochemicals from Trachyspermum ammi.

Extract of de-oiled seeds of Trachyspermum ammi was purified using macroporous resins and the performance of three resins were evaluated to enrich major phytochemical component. A HPLC method has been developed to separate major phytochemical constituents in the crude (CTAE) and partially purified extracts (PTAE). Macroporous resin assisted enrichment and purification suggested XAD-16 as the most efficient (yield 29.8%) followed by XAD-7HP and Diaion HP-20. Concentrated PTAE was subjected to multiple preparative-TLC to afford three compounds, namely, rosmarinic acid-3-O-glucopyranoside (TA-1), kaempferol-(coumaroyl-glucosyl)-rhamnoside (TA-2) and quercetin-3-O-galactoside (TA-3). The structure of these compounds was elucidated from their corresponding spectroscopic characterizations in FT-IR, HR-MS, and partially by 1 H NMR. Total phenolic and flavonoid contents of the extracts were determined. Antioxidant activity by DPPH and ABTS radical scavenging, CUPRAC assays indicated the highest antioxidant potential of CTAE. Among the compounds, TA-1 exhibited the highest scavenging activity in ABTS (IC50 33.41 µg/ml) and DPPH (IC50 69.23 µg/ml), however, relatively lower than CTAE. In vitro anti-candidal activity against virulent strains of Candida spp. revealed C. albicans 4718 as the most susceptible (23.9 µg/ml) to PTAE. PRACTICAL APPLICATIONS: Seeds of Trachyspermum ammi has been extensively investigated for volatile aromatic components of the essential oil. However, the de-oiled seeds have rarely been exploited for potential bioactive phenolics. The present investigation envisaged possible utilization of the de-oiled Trachyspermum seeds for its phenolic constituents, which could be used as natural antioxidant with additional benefits of anticandidal properties. Indeed, macroporous resin assisted enrichment and purification of extracts of T. ammi seeds generate valuable reference compounds, rosmarinic acid-3-O-glucopyranoside, kaempferol-(coumaroyl glucosyl)-rhamnoside, and quercetin-3-O-galactoside.

Anti-Cancer and Anti-Inflammatory Potential of Furanocoumarins from Ammi majus L.

Secondary metabolites have potential benefits to human being. They are used in the food, agricultural and pharmaceutical industries. The secondary metabolite of furanocoumarins from different plant sources is essential in various skin-related ailments. Biologically, these chemicals are isolated from different plants in the Apiaceae, Fabaceae, Rutaceae and Moraceae families. Ammi majus L. is one of the most common plants in the family of Apiaceae with a large quantity of derivatives. The furanocoumarin derivatives defend the plant by fighting external enemies by Systemic Acquired Resistance (SAR). Via suppressing or retarding microbial growth in infected parts, these derivatives, along with SAR, help to alleviate inflammation in the human body. Latest evidence of these compounds has been established in the treatment of cancer, but the mechanism that needs to be elaborated is not yet understood. Recent studies have shown that furanocoumarin derivatives bind to DNA base pairs and block DNA replication. This may be a potential pathway that helps to regulate the growth of cancerous cells. This article reflects on the pharmaceutical data of furanocoumarins and their different mechanisms in these cases.

Combination effect of red light irradiation and Traychspermum ammi essential oil on colorectal cancer cells (SW480).

Colon cancer is the third significant reason for death related to cancers in the world. There are various treatments for colon cancer, which have several side effects. Polyphenol agents are a type of antioxidant in plants that have diverse biological properties, such as anti-cancer effects. Here, we investigate the effect of Trachyspermum ammi essential oil (TEO) and red light irradiation on the colorectal cancer cell line (SW 480). The colorectal cancer cell lines were irradiated at 660 nm for 90 s and then the cells were incubated with different TEO concentrations. In another study, cells initially were treated with various TEO concentrations and then irradiation for 90 s. Effect of TEO and the red light irradiation on viability of the cell, ROS generation, and cell cycle was assessed by MTT and flow cytometry, respectively. The findings demonstrated that early incubation with TEO and then irradiation decreased the SW 480 cells survival more than the early irradiation at 660 nm and then essential oil. In addition, TEO treatment at IC50 concentration in combination with low-level laser irradiation induces ROS generation in SW 480 cells as compared to the dark group. In addition, TEO treatment at IC50 in combination with low-level laser irradiation induces G0/G1 arrest of the cell cycle in SW 480 cells in comparison to the dark group. This study revealed that the Trachyspermum ammi essential oil in combination with low-level laser results in more reduction in survival which leads to ROS generation and cell cycle arrest in SW 480 colorectal cancer cells.

Trachyspermum ammi and Cinnamomum verum as nutraceuticals: Spices rich in therapeutically significant protein tyrosine phosphatases.

Nutraceuticals need special attention as preventive molecules to create a natural barrier against various dreadful diseases like cancer and to regulate metabolism. In the present study, two spices, Trachyspermum ammi and Cinnamomum verum, been identified as excellent Protein Tyrosine Phosphatases (PTPases) sources that play significant role in the regulation of cell signal transduction and developmental processes in plants as well as animals, being lucrative and potential targets for pharmacological modulation. PTPases from both cases were partially purified into 0%-40% and 40%-80% fractions based on ammonium sulfate saturation levels. Fraction (40%-80%) exhibited a purification level of 4.44-fold and 2.86-fold with specific activity of 44.06 and 23.33 U/mg for PTPases from T. ammi and C. verum, respectively. PTPases being found to be thermally stable up to 70°C imply their industrial significance. Kinetic studies showed Km values to be 7.14 and 8.33 mM, whereas the activation energy (Ea ) values were 25.89 and 29.13 kJ/mol, respectively. Divalent cations: Cu2+ , Zn2+ , and Mn2+ acted as inhibitors of PTPases, from both sources. The Ki values of inhibitors varied from 0.014-0.125 mM in the descending order Cu2+  > Zn2+  > Mn2+ and Mn2+  > Cu2+  > Zn2+ for PTPases from T. ammi and C. verum, respectively. The inhibitory effect of sodium metavanadate aligns with prominent PTPase characteristics. In addition to these properties, the thermostability of PTPases from two spices enhances their significance in industries with therapeutically vital products. Although the source of PTPases is culinary spices, further studies are required to establish the utilization of PTPases as nutraceuticals and in therapeutic formulations. PRACTICAL APPLICATIONS: For a healthy lifestyle, awareness needs to be created by humankind towards food habits to minimize illnesses. Numerous studies have explored the consumption of nutraceutical products acts as a natural barrier and immune booster for various human ailments including SARS-COV-2. PTPases play important roles in regulating intracellular signaling and, ultimately, biological function along with their structural features. The importance of PTPases and their inhibitors has been implicated in various diseases like cancer, diabetes, and obesity. Further investigations need to be undertaken to explore the therapeutic properties of PTPases in both in vivo and in vitro for their clinical significance.

Assessment of Conventional Solvent Extraction vs. Supercritical Fluid Extraction of Khella (Ammi visnaga L.) Furanochromones and Their Cytotoxicity.

BACKGROUND: Khella (Ammi visnaga Lam.) fruits (Apiaceae) are rich in furanochromones, mainly khellin and visnagin, and are thus incorporated in several pharmaceutical products used mainly for treatment of renal stones. METHODS: The objective of this study was to compare the yield of khellin and visnagin obtained using different conventional solvents and supercritical fluid extraction (SCFE) with carbon dioxide (containing 5% methanol as co-solvent). Water, acetone and ethanol (30% and 95%) were selected as conventional solvents. RESULTS: Highest extract yield was obtained from 30% ethanol (15.44%), while SCFE gave the lowest yield (4.50%). However, the percentage of furanochromones were highest in SCFE (30.1%), and lowest in boiling water extract (5.95%). HPLC analysis of conventional solvent extracts showed other coumarins that did not appear in supercritical fluid extraction chromatogram due to non-selectivity of solvent extraction. Ammi visnaga extracts as well as standard khellin and visnagin were tested for their cytotoxic activity using sulforhodamine B assay on breast cancer (MCF-7) and hepatocellular carcinoma (Hep G2) cell lines. Results revealed a strong cytotoxic activity (IC50 < 20 µg/mL) for the SCFE and standard compounds (khellin and visnagin) (IC50 ranging between 12.54 ± 0.57 and 17.53 ± 1.03 µg/mL). However, ethanol and acetone extracts had moderate cytotoxic activity (IC50 20-90 µg/mL) and aqueous extract had a weak activity (IC50 > 90 µg/mL). CONCLUSIONS: Thus, supercritical fluid extraction is an efficient, relatively safe, and cheap technique that yielded a more selective purified extract with better cytotoxic activity.

PLGA-Based Nano-Encapsulation of Trachyspermum Ammi Seed Essential Oil (TSEO-PNP) as a Safe, Natural, Efficient, Anticancer Compound in Human HT-29 Colon Cancer Cell Line.

Colorectal cancer is a lethal and commonly diagnosed cancer worldwide. To halt its burden more efficient targeted strategies are needed. Trachyspermum ammi seed essential oil (TSEO) contains several anticancer phytochemicals that maybe more effective via PLGA-based nano-encapsulation. TSEO-PNP nanoparticles were synthesized utilizing evaporation and ultra-sonication-based emulsification methods. Their size, morphology, and stability were defined by DLS, SEM, and surface zeta-potential data, respectively. The TSEO-PNP antioxidant apoptotic, cytotoxic, and antiangiogenic impacts on both cell lines (HT-29 and HUVEC) were studied by FRAP/ABTS, Q-PCR, MTT, and CAM assays, respectively. Moreover, further confirmatory measurements such as AO/EB fluorescent staining and flow cytometry analysis were performed to verify apoptosis. Stable (-32.42 mV) 206.21-nm TSEO-PNP induced apoptosis in the HT-29 cells. Apoptosis was confirmed by significant overexpression of apoptotic genes (Cas-9 and BAX), down-regulation of the anti-apoptotic (BCL-2) gene, fluorescent AO/EB staining, and flow cytometry data following increased TSEO-PNP treatment doses. TSEO-PNP exhibited a meaningful dose- and time-dependent cancer-specific cytotoxic impact on HT-29 cells. The TSEO-PNP has three main anticancer activities on HT-29 colon cancer cells including oxidant reduction, apoptosis induction, and angiogenesis suppression.

Responses of ajowan (Trachyspermum ammi L.) to exogenous salicylic acid and iron oxide nanoparticles under salt stress.

This research with a factorial arrangement was undertaken to investigate physiological responses of ajowan plants to foliar treatment of salicylic acid (1 mM) and nano-Fe2O3 (3 mM) under various salinity levels (0, 4, 8, 12 dS m-1 NaCl, respectively). Rising salinity enhanced sodium and endogenous SA contents, soluble sugars, protein, glycine betaine, proline, antioxidant enzymes activities, ROS generation, and lipid peroxidation, while reduced potassium and iron contents, membrane stability index, leaf water content, leaf pigments, root and shoot biomasses, and seed yield. Application of particularly SA and SA+nano-Fe2O3 alleviated salt toxicity via enhancing K+ uptake, K+/Na+ ratio, Fe content, endogenous level of SA, the activities of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, and polyphenol oxidase), and most of the osmolytes. These changes were resulted in improving membrane stability index, leaf water content, leaf pigments, root and shoot growth, and finally seed yield of plants under moderate and severe salinities. Therefore, these treatments can additively enhance salt tolerance and physiological performance of ajowan through increasing antioxidant capacity, osmolytes, and photosynthetic pigments. Graphical Abstract .