Design of three-component essential oil extract mixture from Cymbopogon flexuosus, Carum carvi, and Acorus calamus with enhanced antioxidant activity.

The development of novel antioxidant compounds with high efficacy and low toxicity is of utmost importance in the medicine and food industries. Moreover, with increasing concerns about the safety of synthetic components, scientists are beginning to search for natural sources of antioxidants, especially essential oils (EOs). The combination of EOs may produce a higher scavenging profile than a single oil due to better chemical diversity in the mixture. Therefore, this exploratory study aims to assess the antioxidant activity of three EOs extracted from Cymbopogon flexuosus, Carum carvi, and Acorus calamus in individual and combined forms using the augmented-simplex design methodology. The in vitro antioxidant assays were performed using DPPH and ABTS radical scavenging approaches. The results of the Chromatography Gas-Mass spectrometry (CG-MS) characterization showed that citral (29.62%) and niral (27.32%) are the main components for C. flexuosus, while D-carvone (62.09%) and D-limonene (29.58%) are the most dominant substances in C. carvi. By contrast, ß-asarone (69.11%) was identified as the principal component of A. calamus (30.2%). The individual EO exhibits variable scavenging activities against ABTS and DPPH radicals. These effects were enhanced through the mixture of the three EOs. The optimal antioxidant formulation consisted of 20% C. flexuosus, 53% C. carvi, and 27% A. calamus for DPPHIC50. Whereas 17% C. flexuosus, 43% C. carvi, and 40% A. calamus is the best combination leading to the highest scavenging activity against ABTS radical. These findings suggest a new research avenue for EOs combinations to be developed as novel natural formulations useful in food and biopharmaceutical products.

Phytochemical, Morphological, and Physiological Variation in Different Ajowan (Trachyspermum ammi L.) Populations as Affected by Salt Stress, Genotype × Year Interaction and Pollination System.

In the present research, 28 populations of ajowan (Trachyspermum ammi L.) were evaluated for agro-morphological traits and essential oil yield in two consecutive years. Then, selected ajowan populations from these two years were used for further morphophysiological and biochemical studies under different salinity levels (control, 60, 90, and 120 mM NaCl). The main components of the oil were thymol (32.7-54.29%), γ-terpinene (21.71-32.81%), and p-cymene (18.74-26.16%). Salt stress caused an increase in essential oil content in the Esfahfo and Qazvin populations. The highest total phenolic and flavonoid contents were found in the Arak population grown in 60 mM NaCl (183.83 mg TAE g-1 DW) and the Yazd population grown in 90 mM NaCl (5.94 mg QE g-1 DW). Moreover, the Yazd population exhibited the strongest antioxidant activity based on DPPH (IC50 = 1566 µg/mL) under 60 mM NaCl and the highest reducing power (0.69 nm) under 120 mM NaCl. The results revealed that low and moderate salt stress improves the phytochemicals of ajowan seeds, which are useful for pharmaceutical and food applications. In this research, some morphological traits, as well as essential oil yield, were evaluated in open pollinated versus self-pollinated plants. As a result, plant height, number of flowering branches, and crown diameter significantly decreased in some populations, while a significant increase was obtained for number of flowers per umbel and seed numbers per umbel. Finally, self-pollination of ajowan might provide new insights for further breeding programs to increase oil or thymol content in ajowan.

The structural characterization and bioactivity assessment of nonspecific lipid transfer protein 1 (nsLTP1) from caraway (Carum carvi) seeds.

BACKGROUND: Carum carvi (caraway) of the Apiaceae family has been used in many cultures as a cooking spice and part of the folk medicine. Previous reports primarily focus on the medicinal properties of caraway seed essential oil and the whole seeds extract. However, no effort has been made to study caraway proteins and their potential pharmacological properties, including nonspecific lipid transfer protein (nsLTP), necessitating further research. The current study aimed to characterize nonspecific lipid transfer protein 1 (nsLTP1) from caraway seed, determine its three-dimensional structure, and analyze protein-ligand complex interactions through docking studies. We also evaluated nsLTP1 in vitro cytotoxic effect and antioxidant capacity. Additionally, nsLTP1 thermal- and pH- stability were investigated. METHODS: Caraway nsLTP1 was purified using two-dimensional chromatography. The complete amino acid sequence of nsLTP1 was achieved by intact protein sequence for the first 20 residues and the overlapping digested peptides. The three-dimensional structure was predicted using MODELLER. Autodock Vina software was employed for docking fatty acids against caraway nsLTP1. Assessment of nsLTP1 cytotoxic activity was achieved by MTS assay, and the Trolox equivalent antioxidant capacity (TAC) was determined. Thermal and pH stability of the nsLTP1 was examined by circular dichroism (CD) spectroscopy. RESULTS: Caraway nsLTP1 is composed of 91 residues and weighs 9652 Da. The three-dimensional structure of caraway nsLTP1 sequence was constructed based on searching known structures in the PDB. We chose nsLTP of Solanum melongena (PDB ID: 5TVI) as the modeling template with the highest identity among all other homologous proteins. Docking linolenic acid with caraway protein showed a maximum binding score of -3.6 kcal/mol. A preliminary screening of caraway nsLTP1 suppressed the proliferation of human breast cancer cell lines MDA-MB-231 and MCF-7 in a dose­dependent manner with an IC50 value of 52.93 and 44.76 µM, respectively. Also, nsLTP1 (41.4 µM) showed TAC up to 750.4 µM Trolox equivalent. Assessment of nsLTP1 demonstrated high thermal/pH stability. CONCLUSION: To the best of our knowledge, this is the first study carried out on nsLTP1 from caraway seeds. We hereby report the sequence of nsLTP1 from caraway seeds and its possible interaction with respective fatty acids using in silico approach. Our data indicated that the protein had anticancer and antioxidant activities and was thermally stable.

Effects of methyl jasmonate elicitation on the carvone and limonene contents, phenolic compounds and antioxidant activity in caraway (Carum carvi L.) callus cultures.

This study investigated the elicitation effects of several methyl jasmonate (MeJ) concentrations (0, 25, 50, and 100 µM) on various biochemical traits of caraway (Carum carvi L.) callus cultures. The 25 µM MeJ concentration yielded the highest callus growth rate (0.57 mm day-1), total flavonols content (2.58 mg QE g-1 FW) and total carotenoids content (0.04 µg g-1 FW), whereas the highest relative fresh weight (75.72%), total phenolics content (76.90 mg GAE g-1 FW), total flavonoids content (58.49 mg QE g-1 FW) and phenylalanine ammonia lyase activity (3.40 nmol cinnamic acid mg-1 h-1 FW) were obtained with the 50 µM MeJ concentration. The highest antioxidant activity through DPPH assay (8.37%) and malondialdehyde content (7.82 µmol g-1 FW) were observed at 100 µM MeJ. The HPLC conducted 21 days post-elicitation revealed a 5.55-fold increase of carvone accumulation (1.83 µg g-1 DW) at 50 µM MeJ and a 2.7-fold increase (0.62 µg g-1 DW) of limonene at 50 µM MeJ. The optimal dosages applied for carvone and limonene accumulation under MeJ elicitation could be used to scale up the economic production of these elite medicinal compounds through caraway callus cultures.

Morphology, physiology, and biochemistry of zinc-stressed caraway plants.

A greenhouse pot experiment was conducted to evaluate the impact of zinc supply (0, 1, and 2 mM Zn as ZnSO4) on morpho-physiological and biochemical parameters of caraway (Carum carvi L.). Exposure to different Zn concentrations for 12 weeks compromised severely all growth parameters (plant height, number of secondary branches, diameter of primary and secondary branches, fresh and dry weight of aerial parts and roots) yield and its components (number of umbels per primary branches and secondary branches; number of umbel per plant; number of seeds per plant; and the weight of 1000 seeds). These manifestations were intimately linked with excessive accumulation of Zn in roots and leaves, alteration of the content of photosynthetic pigments, and extended lipid peroxidation. A manifest increment of proline and soluble sugar content was also observed in response to Zn application. Lipid content in seeds was dropped in Zn-treated plants and the fatty acid profiles were profoundly affected as they were enriched with saturated fatty acids at the expense of unsaturated ones. While improving their oxidative stability as revealed by the reduced values calculated oxidizability and oxidative susceptibility, Zn treatment reduced the lipid nutritional quality of caraway seeds. Moreover, Zn treatment reduced the essential oil yield and its main component carvone while it enhanced the content of its precursor limonene. It also induced alteration of terpene metabolism as revealed in the redirection of the carbon flux to the shikimate/phenylpropanoid pathway resulting in the stimulation of the production of phenolic compounds and their subsequent antioxidant activities.

Phytochemical constitutes and biological activities of essential oil extracted from irradiated caraway seeds (Carum carvi L.).

PURPOSE: Essential oils (EOs) obtained from spices, herbs, and medicinal plants are well known in traditional medicine and are an area of interest due to their various biological activities. Therefore, the present study investigates the chemical composition, phytochemical properties, as well as the biological activity of EOs, recovered from un-irradiated and irradiated (2.5, 5, and 10 kGy) caraway seeds. MATERIALS AND METHODS: Carum carvi L. seeds were irradiated with gamma irradiation at dose levels 2.5, 5, and 10 kGy, then EOs were recovered from all the samples. The chemical composition, phenols, and flavonoids content were evaluated. As well, antimicrobial and antitumor activities against the two cell lines [colorectal adenocarcinoma (Caco-2) and liver cancer (HepG-2)] were investigated. RESULTS: The results indicated the percentage of oil increased by radiation, especially a dose of 10 kGy, which gave the highest percentage (3.50%) compared to the control. Also, the Gas Chromatography/Mass Spectrometry (GC-MS) analysis revealed the presence of 26 compounds in the essential oil extracts. The main constituent of caraway seeds EOs was Carvone followed by Limonene. According to the results, there was an increase in the content of phenols and flavonoids by using gamma rays compared with control, the maximum increase was observed at dose level 10 kGy (13.70 and 7.38 mg/g oil, respectively) followed by 5 kGy (11.20 and 5.86 mg/g oil, respectively). The antioxidant properties of the caraway essential oils were increased by increasing the irradiation dose level (2.5-10 kGy) analyzed by DPPH radical and metal chelating activity. Caraway essential oils have an antimicrobial action against Gram-positive and Gram-negative bacteria as well as fungi. The antimicrobial activity was increased as the irradiation dose was raised and the10 kGy dose was more effective in suppressing the growth of bacteria and fungi. Additionally, the caraway essential oils have anticancer activity against the two cell lines studied; colorectal adenocarcinoma (Caco-2) and liver cancer cell line (HepG-2) as reduced the cell viability and density. CONCLUSION: The 10 kGy dose was more effective for oil yield, phenols, flavonoids, and antioxidant activity as well as antibacterial and antifungal activities. Furthermore, the caraway essential oils indicated anticancer activity against the two cell lines studied; colorectal adenocarcinoma (Caco-2) and liver cancer cell line (HepG-2) as reduced the cell viability and density. So caraway could be considered an important herb with multiple therapeutic uses.

Stimulation of Secondary Metabolites and γ-Terpinene Synthase by Silver Nanoparticles in Callus Cultures of Carum carvi.

Biotechnology and nanotechnology are important tools for understanding biochemical pathways. They can be used efficiently for stimulating and increasing the production of secondary metabolites in medicinal plants. The present study aimed to identify the γ-terpinene synthase gene (CcTPS2) as an effective contributor to the biosynthetic pathway of monoterpenes. The effects of silver nanoparticles (AgNPs; 50 and 100 mg l- 1) and time (24 and 48 h) were examined on secondary metabolites in cell suspension cultures of Carum carvi. This involved the identification, isolation, and sequencing of a partial sequence in the CcTPS2 gene of C. carvi. The genomic sequence of CcTPS2 comprised 292 bp which were organized into two exons (110 and 82 bp) and one intron (100 bp), while the cDNA was 192 bp. In the scale of nucleotides, the CcTPS2 gene showed 96% similarity with the TPS2 gene of Oliveria decumbens. We generated sequence data of the CcTPS2 gene for the first time in this species, thereby enabling further developments in understanding the molecular mechanisms responsible for terpene biosynthesis and other chemical derivatives in C. carvi. The results of GC/MS and GC/FID showed that AgNPs strongly affected the secondary metabolites in cell suspension cultures of C. carvi. According to the results, the AgNPs (50 mg l- 1) increased p-cymene and carvone contents in comparison with the control. The exposure of plants to 100 mg l- 1 AgNPs induced the production of thymol and carvacrol. The results of real-time PCR revealed that the exposure of plants to 100 mg l- 1 AgNPs caused a significant upregulation of CcTPS2 expression for 24 h. These cell suspension cultures were elicited by AgNPs, the application of which proved as an effective method to improve the production of secondary metabolites in vitro.

Low-dose gamma irradiation and pectin biodegradable nanocomposite coating containing curcumin nanoparticles and ajowan (Carum copticum) essential oil nanoemulsion for storage of chilled lamb loins.

The current investigation assessed the effect of pectin (PE) biodegradable nanocomposite coating containing curcumin nanoparticles (CNP) and ajowan (Carum copticum) essential oil nanoemulsion (ANE) combined with low-dose gamma irradiation on microbial, physiochemical, and sensorial qualities of lamb loins during refrigeration conditions. Active coating combined with gamma irradiation reduced the count number of mesophilic and psychrotrophic bacteria, lactic acid bacteria, Enterobacteriaceae; and minimized lipid and protein oxidation changes, total volatile basic nitrogen content, met-myoglobin formation, and color deterioration in the loin samples. The increased shelf-life of lamb loins up to 25 days compared with 5 days assigned for the control group can be associated with the application of ionizing radiation and edible PE coating containing CNP and ANE, which might be due to the synergistic or additive effects of treatments. Overall, as an effective preservation technique, a combination of PE + CNP + ANE and irradiation can be recommended for prolonging the shelf-life of lamb loins during refrigerated storage.

Characterization and Physical and Biological Properties of Tissue Conditioner Incorporated with Carum copticum L.

AIM: One of the main problems in dentistry is the injury caused by the long-term application of an ill-fitting denture. The existence of multiple microorganisms along with the susceptibility of the tissue conditioners to colonize them can lead to denture stomatitis. This study is aimed at developing a tissue conditioner incorporated with Carum copticum L. (C. copticum L.) for the effective treatment of these injuries. MATERIALS AND METHODS: The Carum copticum L. essential oil composition was determined by gas chromatography-mass (GC-mass) spectrometry. The antimicrobial activity of the essential oil against the standard strains of bacterial and fungal species was determined by broth microdilution methods as suggested by the Clinical and Laboratory Standards Institute (CLSI). The physical and chemical properties of the prepared tissue conditioner were investigated by viscoelasticity, FTIR assays, and the release study performed. Furthermore, the antibiofilm activity of the Carum copticum L. essential oil-loaded tissue conditioner was evaluated by using the XTT reduction assay and scanning electron microscopy (SEM). RESULTS: The main component of the essential oil is thymol, which possesses high antimicrobial activity. The broth microdilution assay showed that the essential oil has broad activity as the minimum inhibitory concentration was in the range of 32-128 µg mL-1. The viscoelasticity test showed that the essential oil significantly diminished the viscoelastic modulus on the first day. The FTIR test showed that Carum copticum L. essential oil was preserved as an independent component in the tissue conditioner. The release study showed that the essential oil was released in 3 days following a sustained release and with an ultimate cumulative release of 81%. Finally, the Carum copticum L. essential oil exhibited significant activity in the inhibition of microbial biofilm formation in a dose-dependent manner. Indeed, the lowest and highest amounts of biofilm formation on the tissue conditioner disks are exhibited in the Streptococcus salivarius and Candida albicans by up to 22.4% and 71.4% at the 64 µg mL-1 concentration of C. copticum L. with a statistically significant difference (P < 0.05). CONCLUSION: The obtained results showed that the Carum copticum L. essential oil-loaded tissue conditioner possessed suitable physical, biological, and release properties for use as a novel treatment for denture stomatitis.

Microencapsulated Caraway Essential Oil Affects Initial Growth of Maize Cultivars.

Caraway (Carum carvi L.) essential oil is a candidate for botanical herbicides. A hypothesis was formulated that the sand-applied maltodextrin-coated caraway oil (MCEO) does not affect the growth of maize (Zea mays L.). In the pot experiment, pre-emergence application of five doses of MCEO was tested on four maize cultivars up to the three-leaf growth stage. The morphological analyses were supported by the measurements of relative chlorophyll content (SPAD), two parameters of chlorophyll a fluorescence, e.g., Fv/Fm and Fv/F0, and fluorescence emission spectra. The analyzed MCEO contained 6.5% caraway EO with carvone and limonene as the main compounds, constituting 95% of the oil. The MCEO caused 7-day delays in maize emergence from the dose of 0.9 g per pot (equal to 96 g m-2). Maize development at the three-leaf growth stage, i.e., length of roots, length of leaves, and biomass of shoots and leaves, was significantly impaired already at the lowest dose of MCEO: 0.4 g per pot, equal to 44 g m-2. A significant drop of both chlorophyll a fluorescence parameters was noted, on average, from the dose of 0.7 g per pot, equal to 69 g m-2. Among the tested cultivars, cv. Rywal and Pomerania were less susceptible to the MCEO compared to the cv. Kurant and Podole. In summary, maize is susceptible to the pre-emergence, sand-applied MCEO from the dose of 44 g m-2.

Antimicrobial, Antioxidant, Anti-Acetylcholinesterase, Antidiabetic, and Pharmacokinetic Properties of Carum carvi L. and Coriandrum sativum L. Essential Oils Alone and in Combination.

Herbs and spices have been used since antiquity for their nutritional and health properties, as well as in traditional remedies for the prevention and treatment of many diseases. Therefore, this study aims to perform a chemical analysis of both essential oils (EOs) from the seeds of Carum carvi (C. carvi) and Coriandrum sativum (C. sativum) and evaluate their antioxidant, antimicrobial, anti-acetylcholinesterase, and antidiabetic activities alone and in combination. Results showed that the EOs mainly constitute monoterpenes with γ-terpinene (31.03%), ß-pinene (18.77%), p-cymene (17.16%), and carvone (12.20%) being the major components present in C. carvi EO and linalool (76.41%), γ-terpinene (5.35%), and α-pinene (4.44%) in C. sativum EO. In comparison to standards, statistical analysis revealed that C. carvi EO showed high and significantly different (p < 0.05) antioxidant activity than C. sativum EO, but lower than the mixture. Moreover, the mixture exhibited two-times greater ferric ion reducing antioxidant power (FRAP) (IC50 = 11.33 ± 1.53 mg/mL) and equipotent chelating power (IC50 = 31.33 ± 0.47 mg/mL) than the corresponding references, and also potent activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) (IC50 = 19.00 ± 1.00 mg/mL), ß-carotene (IC50 = 11.16 ± 0.84 mg/mL), and superoxide anion (IC50 = 10.33 ± 0.58 mg/mL) assays. Antimicrobial data revealed that single and mixture EOs were active against a panel of pathogenic microorganisms, and the mixture had the ability to kill more bacterial strains than each EO alone. Additionally, the anti-acetylcholinesterase and α-glucosidase inhibitory effect have been studied for the first time, highlighting the high inhibition effect of AChE by C. carvi (IC50 = 0.82 ± 0.05 mg/mL), and especially by C. sativum (IC50 = 0.68 ± 0.03 mg/mL), as well as the mixture (IC50 = 0.63 ± 0.02 mg/mL) compared to the reference drug, which are insignificantly different (p > 0.05). A high and equipotent antidiabetic activity was observed for the mixture (IC50 = 0.75 ± 0.15 mg/mL) when compared to the standard drug, acarbose, which is about nine times higher than each EO alone. Furthermore, pharmacokinetic analysis provides some useful insights into designing new drugs with favorable drug likeness and safety profiles based on a C. carvi and C. sativum EO mixture. In summary, the results of this study revealed that the combination of these EOs may be recommended for further food, therapeutic, and pharmaceutical applications, and can be utilized as medicine to inhibit several diseases.

Antispasmodic activity and mechanism of action of polyherbal formulation DCD-684 on rabbit jejunum.

Digas colic drops (DCD-684) is a polyherbal formulation containing decoctions of five medicinal plants namely Carum carvi L., Foeniculum vulgare Mill, Mentha arvensis L., Mentha piperita L. and Zingiber officinale Roscoe. These plants have been extensively used in traditional medicine for the treatment of various gastrointestinal diseases including abdominal colic. This study was conducted to determine the spasmolytic effect of DCD-684 (100% v/v) and its individual plant components on isolated rabbit jejunum (in vitro) and their possible mechanism of action. The effects were evaluated on spontaneous and pre-contracted tissues using KCl (80mM) and other contractile agonists including acetylcholine (0.3µM), carbamylcholine (0.3µM), serotonin (10 µM) and histamine (100µM) in the presence and absence of DCD-684. The various concentrations of DCD-684 (0.1-3% v/v) demonstrated spasmolytic effects on both spontaneous (IC50=0.75%) and KCl-induced contractions (IC50=1.6%), respectively. It also inhibited the contractions induced by acetylcholine (IC50=0.45%), carbamylcholine (IC50=0.95%), serotonin (IC50=0.95%) and histamine (IC50=0.87%). The DCD-684 exhibited synergistic effect due to its five plant components suggesting that spasmolytic cascade is probably governed by muscarinic and/or nicotinic receptors, serotonergic histaminergic, as well as calcium channel blocking mechanisms. Thereby, providing the pharmacological basis of its therapeutic use in the gastrointestinal motility disorders and related inflammatory ailments.

Bacterial and oxidative control of local butter with smart/active film based on pectin/nanoclay/Carum copticum essential oils/ß-carotene.

In this study, Pectin/Nanoclay (montmorillonite)/Carum copticum Essential oils/ß-Carotene (Pec/Clay/CCE/ßC) composite film was prepared. The effect of ß-Carotene (ßC) and Carum copticum Essential oils (CCE) concentration on the antioxidant activity and color/mechanical properties of Pec/Clay/CCE/ßC film was studied. The morphology and antibacterial activity of film were studied. The films containing maximum essential oil and ß-carotene showed the best antibacterial activity, antioxidant activity, flexibility and firmness. So the Pec/Clay/CCE0.5%/BC0.03% film as the optimum film was used for packaging of local butter. Oxidative stability, microbial count, and color properties (L*, a* and b*) of butter packaged with active film (Pec/Clay/CCE/ßC) were investigated. Results showed that Pec/Clay/CCE/ßC film was effective against Bacillus cereus (B. cereus) more than Escherichia coli (E. coli). According to the results active packaged butter had the highest oxidative stability, lowest microbial load, and the least color change during storage. In the packaging process, Pec/Clay/CCE/ßC film color was changed from orange to light yellow and this color change was used as a smart color indicator to detect the oxidation of butter and expiration time of butter.

Rapid Spectrophotometric Method for Assessing Hydroperoxide Formation from Terpenes in Essential Oils upon Oxidative Conditions.

Essential oils are widely used in the food and cosmetics industry as natural flavoring and fragrance substances. For this reason, a thorough quality control applying selected analytical methods is required. Oxidation along with hydroperoxide formation is an important drawback during production and storage of essential oils. Hydroperoxides constitute the main products formed upon photo-oxidation of essential oils. Due to hydroperoxide instability, gas chromatography (GC) and high-performance liquid chromatography (HPLC) analyses are required. According to the European Pharmacopoeia, titration is the official method for oxidation assessment. However, this analysis is time-consuming, and large sample quantities are required. Here, we present a simple and accurate spectrophotometric method for the detection of peroxide trace amounts in essential oils and terpenes. The principle is based on the formation of Wurster's red, which is enforced by the peroxide-driven oxidation of N,N-dimethyl-p-phenylenediamine dihydrochloride (DMPD). The method was validated using dibenzoyl peroxide (DBP) and cumene hydroperoxide (CHP). To demonstrate the suitability of the method for routine analysis, various oxidized terpenes and essential oils were chosen. Moreover, photo- and thermal oxidation experiments were compared and evaluated using gas chromatography/mass spectrometry (GC/MS) and a synthesized limonene-2-hydroperoxide (Lim-2-OOH) reference standard to gather detailed information on the structural changes of the respective terpenes.

Comparative Effects of Carum copticum Essential Oil on Bacterial Growth and Shiga-Toxin Gene Expression of Escherichia coli O157:H7 at Abused Refrigerated Temperatures.

Abused refrigerated temperatures are described as unacceptable deviations from the optimal temperature, occurring frequently during transportation of food products. Escherichia coli O157:H7 is a serious contaminant of meats and meat products due to its ability to grow at abused temperatures (> 10 °C). The aim of this study was to evaluate the antibacterial activity of Carum copticum essential oil for the control of Escherichia coli O157:H7 using laboratory media and minced beef at severe abused refrigerated temperature (15 °C). A comparative quantitative reverse transcription real-time PCR was used to assess effects of temperature and Carum copticum essential oil at sub-minimum inhibitory concentrations on bacterial growth and Shiga-toxin gene (stx1A and stx2A) expression. Results indicated that Carum copticum essential oil inhibited growth of E. coli O157:H7 in tryptone soy broth (TSB) media at all sub-MIC values until Hour 48. However, bacterial population increased progressively until Hour 72 at essential oil concentration of 0.75% (ml g-1) and reached 8.6 log CFU g-1 in minced beef. The essential oil at concentration of 0.005% (ml g-1) increased stx gene expression at all times, but increased stx gene expression (0.015%) at Hour 24 in TSB media. The expression rate of stx1A in minced beef decreased progressively (10.39 and 7.67 folds for 0.5 and 0.75%, respectively) and expression of stx2A was variable in minced beef during storage. In conclusion, results from this study have shown that effects of Carum copticum essential oil on growth and virulence gene expression are not necessarily correlated and temperature, essential oil concentration, investigated gene type, and bacterial growth environment (in vivo or in vitro) are effective as well.

Plasticizers and BPA in spices and aromatic herbs of Mediterranean areas.

This research is carried out in order to characterize the actual contamination by two ubiquitous environmental pollutants, plasticizers and Bisphenol A, in spices (black pepper, caraway and coriander) and aromatic herbs (fennel, laurel, mint, oregano, rosemary, thyme and verbena) from Algeria (n = 26), Tunisia (n = 65) and Italy (n = 53). Algerian samples seem to contain fewer residues than Italian and Tunisian samples. Among the Italian samples, only aromatic herbs, precisely mint, oregano, and laurel, were contaminated. In general, all Tunisian samples showed five plasticizers residues: the caraway, among the spices, and the rosemary, among the aromatic herbs, are found to contain more residues. Also, dietary intake of these contaminants by spices and aromatic herbs under analysis seems not to constitute a risk to the consumers.

Changes in beneficial bioactive compounds in eight traditional herbal liqueurs during a one-month maceration process.

BACKGROUND: Herbs are rich in various beneficial bioactive compounds and they can be used for many different purposes. One of the most common is maceration of herbs in alcohol. Different substances respond differently to extraction in prepared solutions. It is very important to enhance the highest proportion of beneficial ingredients during the maceration process in a herbal liqueur. A comparative analysis of numerous different phenolic compounds from eight of the most frequently used herbs for making aperitifs in Europe was performed. RESULTS: In the comparison among all studied herbs, the highest phenolic content was found with Artemisia absinthium (wormwood) (14.61 mg g-1 ) and Mentha piperita (peppermint) (13.89 mg g-1 ), while the lowest content was found with Centaurium erythraea (common centaury) (3.96 mg g-1 ). Salvianolic acid isomers and caffeic acid derivatives were the greatest contributors to the total phenolic content in lemon balm, wormwood, peppermint, fennel and sage. These compounds contain more hydroxyl groups and they were better extracted at the beginning of the maceration process. Caraway and common centaury contain more flavonols (quercetin and kaempferol derivatives), with higher chemical stability and fewer hydroxyl groups in their structure. CONCLUSION: The compositions of eight herbal liqueurs were highly related to the presence of different herbal ingredients and their solubility in the extract over time. Most flavonol derivatives were extracted over a longer time and the two liqueurs (common centaury and caraway) therefore had the highest phenolic contents after 3 weeks of maceration. © 2019 Society of Chemical Industry.

Bioactive extracts of Carum copticum and thymol inhibit biofilm development by multidrug-resistant extended spectrum ß-lactamase producing enteric bacteria.

The emergence and spread of multidrug-resistant (MDR) pathogenic bacteria is a clinical problem that requires novel anti-infective agents. Targeting pathogenic biofilms is considered a promising strategy to control bacterial infections. In this study, bioactive extracts of Carum copticum were investigated for their anti-biofilm efficacy against extended spectrum ß-lactamase (ESßL) producing MDR enteric bacteria. Thymol was also tested for its anti-biofilm properties, as gas chromatography-mass spectrometry revealed a high content (65.8%) of this phytochemical in the C. copticum methanolic extract. Biofilm inhibition was assessed in microtitre plates and further validated by light, electron and confocal laser microscopy. Sub-inhibitory concentrations of bioactive extracts of C. copticum and thymol significantly prevented biofilm development, ranging from 78.6 to 83.9% reductions. Microscopic analysis revealed that biofilms made by ESßL producing MDR enteric bacteria had a weakened structure, scattered microcolonies, and reduced cell density and thickness after exposure to the bioactive extracts and thymol.

Synthesis of Carum Carvi essential oil nanoemulsion, the cytotoxic effect, and expression of caspase 3 gene.

Scientists are attempting to find novel methods to overcome cancers. Nanoemulsion systems as the novel drug delivery tools have been widely used in cancer therapy. In this study, the Carum Carvi oil nanoemulsions (CCONE) were prepared and its cytotoxic activity was studied on human colon cancer HT-29 cells using MTT assay. Flow cytometry and Real-time qPCR were triggered to evaluate the nanoemulsions' apoptotic properties. The results showed a significant negative association between the HT-29 cancer cell viability and CCONE doses of treatments compared with Huvec normal cells (p value < 0.001). The IC50 values were estimated 12.5 µg/ml and 50 µg/ml for HT-29 and Huvec, respectively. Moreover, we observed that increasing concentrations of nanoemulsions significantly upregulate Caspase-3 gene expression. The results showed the CCONE is an efficient novel apoptosis inducer for human colon cancer cells without any undesirable side effects. However, further in vitro and in vivo researches are required. PRACTICAL APPLICATIONS: Cancer is a complex and usually untreatable disorder. Several types of cancer therapy strategies have been applied widely to overcome cancers. Chemotherapy has been used in various types of cancers. In most cases, not only it had not been effective on cancer cells but also been distractive within normal tissues. According to results, Carum Carvi essential oil nanoemulsions have apoptotic and cytotoxic effects on colon cancer cells (HT-29). When it comes to cancer of any kind, it's important to realize that no dietary supplement can fully treat, cure, or prevent cancer. However, there are some supplements that can potentially decrease the risk of cancer. Nanoemulsions present several advantages including the ability to incorporate hydrophilic, amphiphilic, and lipophilic excipient ingredients, high physical stability, and rapid gastrointestinal digestibility. The Carum Carvi essential oil nanoemulsion can also be applied as an effective food supplement due to its potent apoptotic activity.

para-Menthane as a Stable Terpene Derived from Orange By-Products as a Novel Solvent for Green Extraction and Solubilization of Natural Substances.

This study aims at investigating p-menthane, a novel bio-based solvent resulting from the hydrogenation of d-limonene, as a green alternative to n-hexane or toluene for the extraction and solubilization of natural substances. First, conductor-like combination of quantum chemistry (COSMO) coupled with statistical thermodynamics (RS) calculations show a comparable solubilization profile of p-menthane and n-hexane for carotene, volatile monoterpenes such as carvone and limonene, and model triglycerides. Other data obtained experimentally in solid/liquid extraction conditions further indicate that p-menthane showed similar performances to n-hexane for extracting carotenes from carrots, aromas from caraway seeds, and oils from rapeseeds, as these products showed a comparable composition. p-Menthane was also tested using common analytical extraction procedures such as Soxhlet for determination of oil content via multiple extraction stages, and Dean-Stark for determination of water content via azeotropic distillation. For both systems, yields were comparable, but for Dean-Stark, the distillation curve slope was higher when using p-menthane, and the time needed to attain 100% water recovery was 55% shorter than for toluene. Taken together, these results reveal the potential of p-menthane as a green replacer for petroleum-based solvents such as n-hexane or toluene.