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.

The effect of caraway oil-loaded bio-nanoemulsions on the growth and performance of barnyard grass and maize.

A proper formulation is crucial to improve the herbicidal effects of essential oils and their selectivity. In this study, we investigated the physicochemical properties of bio-based nanoemulsions (CNs) containing several concentrations of caraway (Carum carvi) essential oil stabilized with Eco Tween 80, as a surfactant, maintaining 1:1 proportions. Detailed physicochemical characteristics of the CNs revealed that their properties were most desired at 2% of the oil and surfactant, i.e., the smallest droplet size, polydispersity index, and viscosity. The CNs caused biochemical changes in maize and barnyard grass (Echinochloa crus-galli) seedlings, however, to a different extent. Barnyard grass has overall metabolism (measured as a thermal power) decreased by 39-82% when exposed to the CNs. The CNs triggered changes in the content and composition of carbohydrates in the endosperm of both species' seedlings in a dose-response manner. The foliar application of CNs caused significant damage to tissues of young maize and barnyard grass plants. The effective dose of the CN (ED50, causing a 50% damage) was 5% and 17.5% oil in CN for barnyard grass and maize tissues, respectively. Spraying CNs also decreased relative water content in leaves and affected the efficiency of photosynthesis by disturbing the electron transport chain. We found that barnyard grass was significantly more susceptible to the foliar application of CNs than maize, which could be used to selectively control this species in maize crops. However, further studies are needed to verify this hypothesis under field conditions.

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.

More complex than you think: Taxonomic and temporal patterns of plant-pollinator networks of caraway (Carum carvi L.).

Caraway (Carum carvi L.) is a crop species that is gaining in importance in Europe, especially as a condiment and medicinal plant. Here, we present the plant-pollinator network of caraway in a central European agricultural landscape, focusing on two diverse potential pollinator taxa, Diptera: Brachycera (= true flies) and Hymenoptera (sawflies, bees, and wasps). We specifically studied qualitative differences in interactions between the two insect taxa as well as the intraday and intraseasonal variability of the network. Insect and pollen plant species determination was done via morphological identification and DNA (meta)barcoding. In total, 121 species representing 33 families of Hymenoptera and Brachycera were found to carry caraway pollen. These taxa included many nonhoneybee and nonhoverfly species, showing a wide taxonomic breadth of potential pollinators and a higher network complexity than previously anticipated. There are distinct qualitative differences between Brachycera and Hymenoptera networks, suggesting complementary roles of both taxa in the pollination of native and crop plants. Strong intraday differences in potential pollinator diversity make it necessary to collect insects and pollen at different times of the day to compile complete plant-pollinator networks. Intraseasonal analyses of the plant-pollinator network of caraway show the potential of caraway as an important food source for insect species with an activity peak in late summer.

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.

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.

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.

Kummel's disease

Kummel's disease is a crush fracture of a vertebral body caused by a minor trauma, with the most accepted etiology being avascular necrosis. It is more frequent in individuals with risk factors such as osteoporosis or prolonged treatment with corticosteroids. Initially, it usually has normal radiological tests and an asymptomatic period, followed by a progressive onset of pain along with probable kyphosis and a sign of void or fluid abscess on radiological tests, which may create nerve/spinal involvement. The case is presented of a 76-year-old man, who was admitted to this center for the differential diagnosis of a single vertebral lesion. After imaging tests and biopsy, the definitive diagnosis of Kummel's disease was reached. The confirmatory diagnosis was reached by vertebral biopsy, but given its invasive nature, imaging techniques can play a significant role. As regards metabolic imaging tests, bone scintigraphy has shown to be one of the most sensitive tools to detect ischemia in earlier stages or to determine if it affects other locations. The whole body scan with diphosphonates shows an increase in activity in relation to bone remodeling activity in this condition. The 3-phase study makes it possible to differentiate whether it is an acute/subacute or chronic process, and can influence the therapeutic decision. Knowledge of this disease is important to make a differential diagnosis with tumour or infectious pathology, with emphasis on performing imaging tests in the event of persistent pain with a normal initial plaque.

La enfermedad de Kummel es una fractura-aplastamiento de un cuerpo vertebral precedida por un traumatismo menor, cuya etiología más aceptada es la necrosis avascular. Es más frecuente en individuos con factores de riesgo como osteoporosis o tratamiento prolongado con corticoides. Inicialmente, suele presentar pruebas radiológicas normales y un periodo asintomático, con aparición progresiva del dolor junto a probable cifosis y signo del vacío o absceso líquido en las pruebas radiológicas, pudiendo llegar a crear compromiso nervioso/medular. Presentamos el caso de un varón de 76 arios que ingresa en nuestro centro para el diagnóstico diferencial de una lesión única vertebral; tras la realización de las pruebas de imagen y biopsia se llega al diagnóstico definitivo de enfermedad de Kummel. El diagnóstico de confirmación de esta enfermedad se alcanza mediante la biopsia vertebral, pero dada su naturaleza invasiva, las técnicas de imagen toman un papel relevante. En relación con las pruebas de imagen metabólicas, la gammagrafía ósea ha demostrado ser una de las herramientas más sensibles para detectar isquemia en fases más tempranas o para conocer si afecta a otras localizaciones. El rastreo corporal de cuerpo completo con difosfonatos muestra un aumento de actividad en relación con la actividad ósea remodelativa en este cuadro. El estudio de tres fases permite diferenciar si se trata de un proceso agudo/subagudo o crónico, lo que influye en la decisión terapéutica. Es importante el conocimiento de esta enfermedad para realizar diagnóstico diferencial con patología tumoral o infecciosa e insistir en la realización de pruebas de imagen ante la persistencia del dolor con una placa inicial normal.

Adsorption of Carvone and Limonene from Caraway essential oil onto Tunisian montmorillonite clay for pharmaceutical application.

To explore a novel kind of green composite material having excellent antibacterial, antifungal ability and specific-targeting capability for pharmaceutical uses, a novel kind of bio-composite was prepared using sodium purified clay as carrier of Caraway essential oil (CEO). Gas chromatography-mass spectroscopy (GC-MS) analyses of CEO reveals that Carvone (68.30%) and Limonene (22.54%) are the two major components with a minimum inhibitory concentration (MIC) value equal to 125 mg/mL against Staphylococcus (S) aureus bacteria and Candida albicans fungi. Clay from Zaghouan was purified and characterized by X-ray Photoelectron Spectroscopy (XPS), X-ray diffraction (XRD), Fourier transformed infrared spectroscopy (FT-IR) and N2 adsorption-desorption (BET method). Results obtained by chromatograph equipped with a flame ionization detector (GC-FID) show that the concentration of 130 mg/mL of essential oil and 5 h of contact with the purified clay are the optimal conditions for the bio-hybrid formation. The pseudo-second-order model can describe the kinetic study of the adsorption of Carvone and Limonene on sodium montmorillonite, and the adsorption isotherms have been established to the Langmuir type. Limonene registers a maximum adsorption value equal to 3.05 mg/g of clay however Carvone register the higher amount of adsorption (19.98 mg/g) according to its polarity and the abundance of this compound in the crude CEO. X-ray diffraction, Fourier transformed infrared spectroscopy, elemental analyses (CHN) and X-ray fluorescence characterization valid the success adsorption of CEO in sodium montmorillonite surface. The purified clay/CEO hybrid (purified clay/CEO) combined the advantages of both the clay and the essential oil used in exerting the antibacterial and antifungal activity, and thus, the composite has a double antibacterial and antifungal activity compared to the separately uses of inactive clay and CEO, suggesting the great potential application in pharmaceutical treatments.

Wound healing effect of Carum carvi L. on the incised skin wound in male rats: Histopathology, total protein and biomechanical evaluations.

BACKGROUND: The skin has several functions, one of the most important of which is to protect the internal organs from external damage and the entry of germs. Since skin and wound healing is one of the mostly concerned issues worldwide, the development of wound healing remedies is one of the main fields in modern medical research. OBJECTIVES: To evaluate the effect of the hydroalcoholic extract of Cumin carvi L. seed as one of the traditional medicinal plants used for wound healing through an in vivo model. METHODS: Wide circular skin wounds (2 × 2 cm) were created on the dorsal area of 50 Sprague-Dawley male rats ​​following ethical principles. The animals were divided into five groups including no treatment, base gel, tetracycline treatment, 10% v/v hydroalcoholic extract of Cumin carvi L. seed treatment and 20% v/v hydroalcoholic extract of Cumin carvi L. seed treatment group. Treatment was performed within 20 days. On days 1, 3, 5, 7 and 10, photographs were taken, and the percentage of wound healing was calculated. Also, on the 10th day, the skin area was sampled for histopathology and on the 20th day, the skin was sampled for biomechanical and total protein assessments. RESULTS: The results of wound healing percentage showed that from day 3 onwards, there was a significant improvement between the group treated with 20% v/v hydroalcoholic extract of Cumin carvi L. and the negative control and basal gel groups (p < 0.05). According to histopathological and total protein content evaluations, the amount of collagen production and inflammation score in the Cumin carvi-treated groups confirmed the healing process compared to other groups. CONCLUSIONS: According to the results of this project, 20% v/v ethanolic extract of Cumin carvi L. has potential therapeutic effects in the treatment of skin wounds. Wounds are one of the health concerns, and the economic burden of wound care and healing has continued to increase over the past years.  The healing effects of C. carvi. ethanolic extract on the wound were investigated. The histopathological and macroscopical evaluations, as well as total protein content, were measured to investigate its wound healing properties.

Effect of Caraway Seed Extract on the Blood Biochemistry and Antioxidant Capacity among the Hyperoxidative Stress-Induced Rats.

Carum carvi (Carium) or caraway is traditionally used for the treatment of several metabolic and non-metabolic disorders. In the current study, extracted oil, flavonoids, and alkaloids from the Carium were used to evaluate the effects of these components on blood lipid profile and heart regeneration from oxidative damages caused by hydrogen peroxide consumption. A total of 50 male BALB/c mice were used in this study with a body weight of 23-32 g. The animals were randomly divided into 5 groups (n=10). Group 1: The animals in this group were considered the control group and fed with a normal diet. Group 2: Hyperoxidative stress was induced in this group by giving hydrogen peroxide at a concentration of 1% into the drinking water for 6 weeks. After this period, they did not receive any treatments and only received saline solution by intraperitoneal (IP) injection once a day for 4 weeks. Group 3: Hyperoxidative stress was induced by hydrogen peroxide at a concentration of 1% for 6 weeks. All the animals in this group received 1.25 mg/kg body weight (B.W.) extracted oil from Caraway seeds for 4 weeks by IP injection once a day each week. Group 4: Hyperoxidative stress was induced by hydrogen peroxide at a concentration of 1% for 6 weeks. All the animals in this group received 61.28 mg/kg B.W. extracted flavonoids from Caraway seeds for 4 weeks by IP injection once a day each week. Group 5: Hyperoxidative stress was induced by hydrogen peroxide at a concentration of 1% for 6 weeks. All the animals in this group received 7.8 mg/kg B.W. extracted alkaloids from Caraway seeds for 4 weeks by IP injection once a day each week. The levels of glutathione and malondialdehyde were estimated in the liver and kidneys in the animals with cardiovascular disorders induced by hydrogen peroxide at a concentration of 1%. The results of the current study showed that the alkaloids had the greatest effect in reducing harmful total cholesterol and a complete recovery of the heart and aorta from atherosclerotic lesions through viewing the tissue sections.

Ion contents, physiological characteristics and growth of Carum copticum as influenced by salinity and alkalinity stresses.

A controlled experiment was conducted to investigate the effect of salinity and alkalinity stresses on the growth and physiological characteristics of Carum copticum L. The treatments included four salinity levels: 0, 50, 100 and 150 mM NaCl, and four alkalinity levels 0, 20, 40 and 60 mM NaHCO3. The results indicated that root dry weight and magnesium concentration were decreased and catalase and peroxidase activity, proline, malondialdehyde, Na+ and Ca2+ concentration were increased in plants simultaneously subjected to both salinity and alkalinity stresses. In all traits, the highest salinity and alkalinity levels had the most negative and significant effects. In general, our findings revealed that alkalinity and salinity stresses considerably decreased ajwain growth through adverse impact on physiological characteristics such as ion concentration and activity of antioxidant enzymes. These effects were greater when the two salinity and alkalinity stresses were simultaneously applied. Ajwain plant tolerated a part of the stresses via osmotic adjustment mechanism was assessed by proline, malondialdehyde and total carbohydrate.

How the Past Shapes the Future and What to Do About It: A Discussion at CI 2021 Virtual With Panelists: Teresa Caraway, Ph.D., C.C.C.-S.L.P.; Cliff Megerian, M.D.; Debara Tucci, M.D., M.S., M.B.A.; Peter Thomas, J.D.; Jason Wigand, Au.D.

ABSTRACT: This report summarizes the highlights of a lively discussion between leaders in the cochlear implant (CI) field who also bring significant leadership experience at the practice and health system levels, as well as policy and research perspectives. The discussion revealed several areas in which established patterns of practice should be reconsidered to facilitate wider acceptance of the CI as part of an integrated continuum of hearing healthcare. The importance of continued innovation to advance outcomes, lower cost, and increase access was emphasized. Trends in healthcare funding, which increasingly reward quality at lower cost, will also challenge longstanding assumptions that have influenced program design. Proactive efforts that pursue consistent value in the episode of surgical care while establishing innovative delivery models for audiological and rehabilitation services are the best hope for a future of equitable access and expanding impact.

Phytochemical Analysis and Antioxidant, Antibacterial, and Antifungal Effects of Essential Oil of Black Caraway (Nigella sativa L.) Seeds against Drug-Resistant Clinically Pathogenic Microorganisms.

Nigella sativa (NS) is a plant that has long been utilized in traditional medicine as a treatment for certain diseases. The aim of this work was to valorize the essential oil (EO) of this species by phytochemical analysis and antimicrobial and antioxidant evaluation. EO was extracted by hydrodistillation from the seeds of Nigella sativa (EO-NS). Phytochemical content of EO-NS was evaluated by use of gas chromatography coupled to mass spectrometry (GC-MS/MS). Antioxidant ability was in vitro determined by use of three assays: 2.2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing power (FRAP), and total antioxidant capacity (TAC) relative to two synthetic antioxidants: BHT and quercetin. Antimicrobial effect was evaluated against four clinically important bacterial strains (Staphylococcus aureus, ATCC 6633; Escherichia coli, K12; Bacillus subtilis, DSM 6333; and Proteus mirabilis, ATCC 29906) and against four fungal strains (Candida albicans, ATCC 10231; Aspergillus niger, MTCC 282; Aspergillus flavus, MTCC 9606; and Fusarium oxysporum, MTCC 9913). Fifteen constituents that accounted for the majority of the mass of the EO-NS were identified and quantified by use of GC-MSMS. The main component was O-cymene (37.82%), followed by carvacrol (17.68%), α-pinene (10.09%), trans-sabinene hydrate (9.90%), and 4-terpineol (7.15%). EO-NS exhibited significant antioxidant activity with IC50, EC50, and total antioxidant capacity (TAC) of 0.017 ± 0.0002, 0.1196 ± 0.012, and 114.059 ± 0.97 mg EAA/g, respectively. Additionally, EO-NS exhibited promising antibacterial activity on all strains under investigation, especially on E. coli K12 resulting in inhibition diameter of 38.67 ± 0.58 mm and a minimum inhibitory concentration (MIC) of 1.34 ± 0.00 µg/mL. Also, EO-NS had significant antifungal efficacy, with a percentage of inhibition of 67.45 ± 2.31% and MIC of 2.69 ± 0.00 µg/mL against F. oxysporum, MTCC 9913 and with a diameter of inhibition 42 ± 0.00 mm and MIC of 0.67 ± 0.00 µg/mL against C. albicans. To minimize development of antibiotic-resistant bacteria, EO-NS can be utilized as a natural, alternative to synthetic antibiotics and antioxidants to treat free radicals implicated in microbial infection-related inflammatory reactions.

Caraway (Carum carvi L.) in fast-growing and slow-growing broiler chickens' diets and its effect on performance, digestive tract morphology and blood biochemical profile.

The aim of this study was to evaluate the addition of caraway (1%) in fast-growing and slow-growing broiler chickens' diet and its effect on performance parameters, blood biochemical profile, and relative organ sizes and ileum morphology in slow-growing broilers. Two separated experiments were performed. On the first day of age, the broilers were divided into 2 equal groups (Control and Caraway) with 6 replicates per treatment in both experiments. Experiment I: The total of 276 male fast-growing Ross 308 broiler chickens were used. The trial lasted from the first day to 35th day of chickens' age. Experiment II: The total of 216 male slow-growing (Hubbard JA 57) broilers were used. The trial lasted from the first to 50th day of chickens' age. Mean liveweight, weight gain, feed conversion ratio, blood biochemical parameters, and relative organ sizes were not significantly different in these trials. The group of slow-growing broilers supplemented with 1% of caraway in the diet showed longer villi and deeper crypt in the ileum after 50 d of life. Based on our results, it can be stated that the proportion of 1% caraway in fast-growing and slow-growing broiler chickens' diet did not influence performance parameters, blood biochemical profile and relative organ sizes. In case of the experiment with the slow-growing broilers supplemented with caraway, a significant difference in the height of the villi and the depth of the crypts was found. Caraway can be included in the broiler chickens' diets without negative effects, but further study of the effect on the intestinal morphology is necessary.

The influence of Nigella sativa essential oil on proliferation, activation, and apoptosis of human T lymphocytes in vitro.

In previous work, we tested the immunomodulatory effect of Nigella sativa (NS) fatty oil. Our results demonstrated that unrefined, obtained by cold pressing black cumin seed oil inhibited lymphocytes' proliferation and induced their apoptosis in a dose-dependent manner. In this study, we examined the immunomodulatory properties of essential oil (EO) obtained from the NS seeds by hydrodistillation and its two main constituents: thymoquinone (TQ) and p-cymene. We analyzed the proliferation, activation phenotype, and apoptosis rates of human T lymphocytes stimulated with an immobilized monoclonal anti-CD3 antibody in the presence of serial ethanol dilutions of tested oil or serial distilled water dilutions of tested compounds with flow cytometry. Our results showed that NSEO significantly inhibited the proliferation of CD4+ and CD8+ T lymphocytes, induced cell death in a dose-dependent manner, and reduced the expression of CD28 and CD25 antigens essential for lymphocyte activation. TQ inhibited the proliferation of T lymphocytes and induced cell death, particularly in high concentrations. Meanwhile, p-cymene did not influence lymphocyte proliferation. However, its high concentration induced cell necrosis. These results show that the essential oil from Nigella sativa has powerful immunomodulatory properties, which, at least partially, are related to the TQ component.

Chemical profiling of composite prescription caraway and quantification of three pairs isomeric components in caraway administered rat plasma by tandem mass spectrometry.

Caraway, a well-known traditional Uyghur medicine, has been used to treat vitiligo for centuries. Its biological effects on melanin synthesis of caraway have been investigated. However, beyond psoralen and isopsoralen alone, no further chemical component of caraway has been revealed. In this study, ultra-high performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry was employed to comprehensively characterize the chemical components present in caraway. Based on accurate mass measurements, key fragmental ions and comparison with reference standards, 75 chemical components were identified in caraway. Moreover, a tandem mass spectrometry method was developed and validated for quantitative analysis of three pairs isomeric components, namely psoralen/isopsoralen, bavachin/isobavachalcone and bavachromene/isobavachromene in rat plasma. Psoralen, isopsoralen, bavachin, and isobavachalcone showed linearity with concentration ranging of 1.0-500.0 ng/ml. The linear ranges for bavachromene and isobavachromene were 0.2-500.0 ng/ml. The accuracies were in ranges of 85%-115% with coefficient of variation errors of less than 15%. Furthermore, the method was applied to quantify the three pairs isomeric components in rats after oral administration of caraway.

Synergistic anti-cancer effects of Nigella sativa seed oil and conventional cytotoxic agent against human breast cancer.

OBJECTIVES: Breast cancer is the most commonly diagnosed invasive non-skin malignancy in women worldwide, and it is the leading cause of cancer-related deaths in them. Nigella sativa Linn. seed oil has been found to be effective in cancer treatment as well as having anti-cancer properties in some other types of cancers. The study looked into the synergistic cytotoxic effects of N. sativa Linn. seed oil and doxorubicin in the treatment of human breast cancer cells (MCF-7). METHODS: Nigella sativa Linn. seed oil was used to evaluate its effect on human breast cancer cells, either alone or in conjunction with doxorubicin. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tests were used to examine cell proliferation and cell viability, while phase-contrast inverted microscopy was used to examine cellular morphology. Furthermore, the role of N. sativa seed oil in decreasing cell tumorigenicity features was highlighted by testing the cancer cell migration using the wound healing assay. RESULTS: Results showed that higher concentrations (50 µg/mL) of N. sativa Linn. seed oil changed the breast cancer cell morphology and decreased the cell proliferation and viability. Breast cancer cells treated with black seed oil decreased cell movement after 24 hours compared to the untreated cell in the wound healing assay. Whereas, only the higher concentration of doxorubicin (0.5-2.5 µg/mL) reduced cell proliferation and cell viability. Moreover, the combination treatment of 50 µg/mL of black seed oil with different concentrations of doxorubicin caused a significant cell proliferation reduction and decreased cell viability. The activity was seen optimum at lower concentration (0.1 µg/mL) of doxorubicin. CONCLUSIONS: There was decreased cell proliferation and cell viability when N. sativa seed oil was used alone or in conjunction with doxorubicin in Breast cancer cells (MCF-7) revealing potential opportunities in the field of cancer treatment.

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.