Chemical Composition, Antioxidant, and Antibacterial Activity of Ruta chalepensis L. Ethanolic Extract.

Ruta chalepensis L. is a versatile herb used in culinary arts and traditional medicine. The study aimed to determine the chemical composition of an ethanolic extract from R. chalepensis and the total phenolic and flavonoid content. Additionally, the extracts' antimicrobial and antioxidant activities were tested. The disc diffusion method and minimum inhibitory concentration (MIC) were used to test the antibacterial properties on four types of bacteria: Escherichia coli, Proteus penneri, Bacillus cereus, and Staphylococcus aureus. A colorimetric assay was used to evaluate the total phenolic and flavonoid content, and the DPPH method was used to assess the antioxidant activity. The phytochemical constituents were determined using LC-MS/MS. The results indicated that R. chalepensis ethanolic extract had 34 compounds, and the predominant compounds were quercetin (9.2 %), myricetin (8.8 %), and camphene (8.0 %). Moreover, the extract had a good level of polyphenols and flavonoids, as demonstrated by inhibiting free radicals (DPPH) (IC50 was 41.2±0.1). Also, the extract exhibited robust antimicrobial activity against P. penneri and S. aureus with an MIC of 12.5 and 25.0 µg/mL, respectively. In conclusion, the results suggest that the R. chalepensis ethanolic extract has good antioxidant and antibacterial properties that could be utilized to develop new antibacterial agents.

(+)-/(-)-Rutabenzofuran A and (+)-/(-)- Rutabenzofuran B: Two unprecedented pairs of Z/E isomeric benzofuran enantiomers from the aerial part of Ruta graveolens L.

Two pairs of Z/E isomeric benzofuran enantiomers possessing unprecedented carbon skeletons featuring ring cleavage and addition reactions in the α-pyrone ring of furocoumarin, named rutabenzofuran A [(+)-1 and (-)-1], and rutabenzofuran B [(+)-2 and (-)-2], respectively, were isolated as minor compounds from the water extract of the aerial part of Ruta graveolens L. Their structures were determined by extensive spectroscopic data analysis. The absolute configurations were assigned by comparing the optical rotation with previous research and the experimental circular dichroism (CD) spectra with the calculated electronic CD (ECD) spectra. (-)-1, (+)-2, and (-)-2 were evaluated for antibacterial, anticoagulant, anticancer, and acetylcholinesterase (AChE) inhibitory activities. No anticancer or anticoagulant activities were observed, yet (-)-2 exhibited weak antibacterial activity against Salmonella enterica subsp. Enterica. At the same time, (-)-1, (+)-2, and (-)-2 displayed weak inhibitory activity on AChE.

Phenolic Content and in Vitro Antioxidant, Anti-Inflammatory and antimicrobial Evaluation of Algerian Ruta graveolens L.

Plants constitute a valuable source of natural antioxidants such as polyphenols and are responsible for exhibiting many biologically significant functions. Ruta species including Ruta chalepensis L. and Ruta graveolens L. are widespread species in Algeria and are used as medicinal plants to treat various diseases; however, so far, most of the conducted studies focused on analyzing alkaloids and essential oils mostly on R. chalepensis. The aim of the present research is to investigate the phenolic profile of the aerial parts of Ruta graveolens L. from Algeria and assess its in vitro antioxidant, anti-inflammatory and antimicrobial properties. The total polyphenols and flavonoids were assessed using colorimetric methods, and the individual polyphenols were identified and quantified using HPLC-DAD-ESI-MS. The antioxidant activity was evaluated with DPPH and ß-carotene tests, and the anti-inflammatory activity with inhibition of bovine serum albumin denaturation and HRBC membrane stabilization methods. The results showed that Ruta graveolens extract is rich in phenolic compounds with a total phenol and flavonoid contents of 41.63±0.394 mg GAE/gE and 13.97±0.33 mg EQ/gE, respectively. Nine phenolic compounds were determined, including three phenolic acids and six flavonoids. Rutin was the major phenolic compound in Ruta graveolens (464.95 µg/g), followed by syringic acid (179.74 µg/g), and naringenin (109.78 µg/g). R. graveolens phenolic extract also showed good antioxidant activity with values of 0.77 mM TE/g DW and 0.37 mM ß-CE/g DW with DPPH and ß-carotene tests, respectively. For the anti-inflammatory activity, the highest tested concentration (200 µg/mL) gave 50.61 % of inhibition of the denaturation of albumin and 44.12 % of membrane stabilization. With regards to antimicrobial results, Staphylococcus aureus was the most sensitive bacteria with an inhibition zone of 14.37 mm and MIC value of 0.625 mg/mL, followed by Listeria monocytogenes (11.75 mm and MIC=1.25 mg/mL), and Escherichia coli (10.25 mm and MIC=1.25 mg/mL).

Ruta graveolens, Peganum harmala, and Citrullus colocynthis methanolic extracts have in vitro protoscolocidal effects and act against bacteria isolated from echinococcal hydatid cyst fluid.

Echinococcosis is a common and endemic disease that affects both humans and animals. In this study, the in vitro activities of methanolic extracts of Ruta graveolens, Peganum harmala aerial parts, and Citrullus colocynthis seeds against protoscolosis and isolated bacterial strains from hydatid cysts were assessed using disc diffusion methods and Minimum Inhibitory Concentration (MIC). The chemical composition of three methanolic extracts was studied using LC-MS. After 3 h of exposure to 40 mg/mL R. graveolens extract, a tenfold protoscolocidal effect was seen when compared to the convintional medication (ABZ) for the same duration (P < 0.05). The bacteria listed below were isolated from hydatid cyst fluid collected from a variety of sick locations, including the lung and liver. Micrococcus spp., E. coli, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter amnigenus, Pseudomonas aeruginosa, Staphylococcus xylosus, and Achromobacter xylosoxidans are among the bacteria that have been identified. The most effective extract was R. graveolens, followed by P. harmala and C. colocynthis, according to the results of antibacterial activity using the disc diffusion method. R. graveolens extract had the lowest MIC values (less than 2 mg/mL) against all microorganisms tested. This shows that the R. graveolens extract has additional properties, such as the ability to be both scolocidal and bactericidal. Because these bacteria are among the most prevalent pathogenic bacteria that increase the risk of secondary infection during hydatid cysts, the results of inhibitory zones and MICs of the R. graveolens methanol extract are considered highly promising.

Chemical constituents of Ruta graveolens L. and their melanogenic effects and action mechanism.

Ruta graveolens L. has been widely used to treat various skin ailments, especially vitiligo. In this study, we isolated a new furanocoumarin named Rutagrarin (1) along with 14 known compounds (2-15) from the aerial parts of R. graveolens and elucidated their chemical structures via various spectroscopy. We found that compound 5 promoted melanogenesis and tyrosinase activity in B16 cells. Further investigation on underlying mechanisms revealed that compound 5 activated the transcription of microtia-related transcription factors and promoted the production of melanin in B16 cells via the Akt/GSK-3ß/ß-catenin pathway. Therefore, we confirmed the traditional efficacy of R. graveolens and speculated that compound 5 could be used as a natural drug to treat vitiligo.

Phytochemical and antioxidant analysis of medicinal and food plants towards bioactive food and pharmaceutical resources.

Plants with medicinal properties play an increasingly important role in food and pharmaceutical industries for their functions on disease prevention and treatment. This study characterizes the phenolic composition and antioxidant activity of seven medicinal and food plants, including the leaves of Salvia officinalis L., Rosmarinus officinalis L., Olea europaea L., and Punica granatum L., as well as the leaves and young stems of Ruta graveolens L., Mentha piperita L., and Petroselinum crispum, Mill., by using colorimetric, chromatographic, and spectrophotometric assays. Results revealed that the hydro-methanolic leaf extracts of P. granatum (pomegranate) displayed the highest content of total phenols (199.26 mg gallic acid per gram of plant dry weight), ortho-diphenols (391.76 mg gallic acid per gram of plant dry weight), and tannins (99.20 mg epicatechin per gram of plant dry weight), besides a higher content of flavonoids (24 mg catechin per gram of plant dry weight). The highest antioxidant capacity measured by ABTS, DPPH, and FRAP (2.14, 2.27, and 2.33 mM Trolox per gram of plant dry weight, respectively) methods was also obtained in pomegranate leaf extracts, being 4-200 times higher than the other species. Such potent antioxidant activity of pomegranate leaves can be ascribed to the presence of different types of phenolic compounds and the high content in tannins, whilst phenolic acids and flavonoids were found to be the dominant phenolic classes of the other six plants. Consequently, despite the well-known antioxidant properties of these plant species, our study suggests pomegranate leaf can stand out as a relatively more valuable plant source of natural bioactive molecules for developing novel functional food-pharma ingredients, with potential for not only promoting human health but also improving bio-valorization and environment.

Chalepin and Chalepensin: Occurrence, Biosynthesis and Therapeutic Potential.

Dihydrofuranocoumarin, chalepin (1) and furanocoumarin, chalepensin (2) are 3-prenylated bioactive coumarins, first isolated from the well-known medicinal plant Ruta chalepensis L. (Fam: Rutaceae) but also distributed in various species of the genera Boenminghausenia, Clausena and Ruta. The distribution of these compounds appears to be restricted to the plants of the family Rutaceae. To date, there have been a considerable number of bioactivity studies performed on coumarins 1 and 2, which include their anticancer, antidiabetic, antifertility, antimicrobial, antiplatelet aggregation, antiprotozoal, antiviral and calcium antagonistic properties. This review article presents a critical appraisal of publications on bioactivity of these 3-prenylated coumarins in the light of their feasibility as novel therapeutic agents and investigate their natural distribution in the plant kingdom, as well as a plausible biosynthetic route.

Anti-MRSA Constituents from Ruta chalepensis (Rutaceae) Grown in Iraq, and In Silico Studies on Two of Most Active Compounds, Chalepensin and 6-Hydroxy-rutin 3',7-Dimethyl ether.

Ruta chalepensis L. (Rutaceae), a perennial herb with wild and cultivated habitats, is well known for its traditional uses as an anti-inflammatory, analgesic, antipyretic agent, and in the treatment of rheumatism, nerve diseases, neuralgia, dropsy, convulsions and mental disorders. The antimicrobial activities of the crude extracts from the fruits, leaves, stem and roots of R. chalepensis were initially evaluated against two Gram-positive and two Gram-negative bacterial strains and a strain of the fungus Candida albicans. Phytochemical investigation afforded 19 compounds, including alkaloids, coumarins, flavonoid glycosides, a cinnamic acid derivative and a long-chain alkane. These compounds were tested against a panel of methicillin-resistant Staphylococcus aureus (MRSA) strains, i.e., ATCC 25923, SA-1199B, XU212, MRSA-274819 and EMRSA-15. The MIC values of the active compounds, chalepin (9), chalepensin (10), rutamarin (11), rutin 3'-methyl ether (14), rutin 7,4'-dimethyl ether (15), 6-hydroxy-rutin 3',7-dimethyl ether (16) and arborinine (18) were in the range of 32-128 µg/mL against the tested MRSA strains. Compounds 10 and 16 were the most active compounds from R. chalepensis, and were active against four out of six tested MRSA strains, and in silico studies were performed on these compounds. The anti-MRSA activity of compound 16 was comparable to that of the positive control norfloxacin (MICs 32 vs 16 µg/mL, respectively) against the MRSA strain XU212, which is a Kuwaiti hospital isolate that possesses the TetK tetracycline efflux pump. This is the first report on the anti-MRSA property of compounds isolated from R. chalepensis and relevant in silico studies on the most active compounds.

Packham's Triumph Pears (Pyrus communis L.) Post-Harvest Treatment during Cold Storage Based on Chitosan and Rue Essential Oil.

Pears (Pyrus communis L.) cv. Packham's Triumph are very traditional for human consumption, but pear is a highly perishable climacteric fruit with a short shelf-life affected by several diseases with a microbial origin. In this study, a protective effect on the quality properties of pears was evidenced after the surface application of chitosan-Ruta graveolens essential oil coatings (CS + RGEO) in four different concentrations (0, 0.5, 1.0 and 1.5 %, v/v) during 21 days of storage under 18 °C. After 21 days of treatment, a weight loss reduction of 10% (from 40.2 ± 5.3 to 20.3 ± 3.9) compared to the uncoated pears was evident with CS + RGEO 0.5%. All the fruits' physical-chemical properties evidenced a protective effect of the coatings. The maturity index increased for all the treatments. However, the pears with CS + RGEO 1.5% were lower (70.21) than the uncoated fruits (98.96). The loss of firmness for the uncoated samples was higher compared to the coated samples. The pears' most excellent mechanical resistance was obtained with CS + RGEO 0.5% after 21 days of storage, both for compression resistance (7.42 kPa) and force (22.7 N). Microbiological studies demonstrated the protective power of the coatings. Aerobic mesophilic bacteria and molds were significantly reduced (in 3 Log CFU/g compared to control) using 15 µL/mL of RGEO, without affecting consumer perception. The results presented in this study showed that CS + RGEO coatings are promising in the post-harvest treatment of pears.

Synergistic Effect of Biosynthesized Silver Nanoparticles and Natural Phenolic Compounds against Drug-Resistant Fish Pathogens and Their Cytotoxicity: An In Vitro Study.

Fish pathogens causing disease outbreaks represent a major threat to aquaculture industry and food security. The aim of the presented study is to develop safe and effective bioactive agents against two bacterial isolates: Aeromonas hydrophila and Pseudomonas fluorescens. We employed a broth microdilution method to investigate the antibacterial effect of biosynthesized silver nanoparticles (AgNPs); rutin, a natural flavonoid extracted from Ruta graveneoles; and heliomycin, a secondary metabolite produced by marine actinomycetes AB5, as monotherapeutic agents. Moreover, AgNPs in combination with rutin (AgNP + R) and heliomycin (AgNPs + H) were examined for their synergistic effect. The cytotoxic effect of individual bioactive compounds and in combination with AgNPs was investigated on epithelioma papulosum cyprini (EPC) fish cell lines. Individual treatment of AgNPs, rutin, and heliomycin exhibited a dose-dependent antimicrobial activity against A. hydrophila and P. fluorescens. Rutin minimum inhibitory concentration (MIC) showed the lowest cytotoxicity when tested on EPC cell lines, while heliomycin MIC was highly cytotoxic. Combined subtherapeutic doses of AgNPs + R and AgNPs + H displayed additive and synergistic effects against A. hydrophila and P. fluorescens, respectively, with improved results and relative safety profile. The study findings demonstrate that a combination of AgNPs and natural bioactive compounds may represent novel therapeutics fighting fish pathogens potentially affecting the fish farming industry.

Genus Ruta: A natural source of high value products with biological and pharmacological properties.

ETHNOPHARMACOLOGICAL RELEVANCE: Ruta genus is constituted by ten species, of which the most commonly described are R. chalepensis and R. graveolens. Ruta plants are perennial shrubs belonging to the family Rutaceae, which are traditionally used in folk medicine, since ancient times mostly for the treatment of various ailments of the womb. AIM OF THE STUDY: To provide a review of the different uses of Ruta species in traditional medicine, as well as, on their multifactorial biological and pharmacological properties. MATERIAL AND METHODS: A search of the literature on genus Ruta and Ruta species was performed using various scientific databases and search engines and the information of articles were reviewed and compiled. RESULTS: Different parts of the plants belonging to Ruta genus are used in folk medicine to treat a wide range of different diseases. The principal use of these is in gynaecological field, but the treatment of pain, fever, nausea, inflammation, infections, nervous disorders, among others, are also described. These plants have been used to fertility regulation, as anti-fertility agent, to control menstrual flux and bleedings, as abortifacient and as contraceptive. The phytochemical composition of these plants consists mainly in essential oil (EO), but phenolic compounds were also reported, like coumarins and flavonoids, as well as alkaloids. Ruta species products like extracts and EOs have shown broad pharmacological activities, such as antimicrobial and antifungal activities, as well as, antiviral and antiparasitic. Moreover, Ruta plants products present antioxidant, neuroprotective, anti-inflammatory, anti-cancer and anti-diabetic activities and demonstrated contraceptive and abortifacient effects. These plants were also tested to be used for non-therapeutic approaches, as bio-insecticides in the control of different insect pests showing to be able to reduce infestation. CONCLUSIONS: Ruta species could be a potential source of natural products with biological activities. Ruta extracts, essential oils and isolated compounds have shown a diverse potential for use in the treatment of different diseases, as well as, for pests control, contributing to the valorisation of these plants. Nonetheless, this review indicates that more studies are needed to demonstrate the full potential of Ruta species, and to further explore the toxicology and safety of these plants.

Synthesis, Characterization, and Histological Evaluation of Chitosan-Ruta Graveolens Essential Oil Films.

The development of new biocompatible materials for application in the replacement of deteriorated tissues (due to accidents and diseases) has gained a lot of attention due to the high demand around the world. Tissue engineering offers multiple options from biocompatible materials with easy resorption. Chitosan (CS) is a biopolymer derived from chitin, the second most abundant polysaccharide in nature, which has been highly used for cell regeneration applications. In this work, CS films and Ruta graveolens essential oil (RGEO) were incorporated to obtain porous and resorbable materials, which did not generate allergic reactions. An oil-free formulation (F1: CS) and three different formulations containing R. graveolens essential oil were prepared (F2: CS-RGEO 0.5%; F3: CS+RGEO 1.0%; and F4: CS+RGEO 1.5%) to evaluate the effect of the RGEO incorporation in the mechanical and thermal stability of the films. Infrared spectroscopy (FTIR) analyses demonstrated the presence of RGEO. In contrast, X-ray diffraction (XRD) and differential scanning calorimetry (DSC) analysis showed that the crystalline structure and percentage of CS were slightly affected by the RGEO incorporation. Interesting saturation phenomena were observed for mechanical and water permeability tests when RGEO was incorporated at higher than 0.5% (v/v). The results of subdermal implantation after 30 days in Wistar rats showed that increasing the amount of RGEO resulted in greater resorption of the material, but also more significant inflammation of the tissue surrounding the materials. On the other hand, the thermal analysis showed that the RGEO incorporation almost did not affect thermal degradation. However, mechanical properties demonstrated an understandable loss of tensile strength and Young's modulus for F3 and F4. However, given the volatility of the RGEO, it was possible to generate a slightly porous structure, as can be seen in the microstructure analysis of the surface and the cross-section of the films. The cytotoxicity analysis of the CS+RGEO compositions by the hemolysis technique agreed with in vivo results of the low toxicity observed. All these results demonstrate that films including crude essential oil have great application potential in the biomedical field.

Composición química y caracterización de flavonoides de extractos metanólicos de hojas de dos tipos de Ruta chalepensis L / Chemical composition and flavonoids characterization of the methanolic extract of leaves of two types of Ruta Chalepensis L

Objetivos. Detectar metabolitos secundarios y caracterizar estructuras químicas de flavonoides de los extractos metanólicos de hojas de dos tipos de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Materiales y métodos. Se elaboraron extractos metanólicos de hojas de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Se llevó a cabo la prueba de solubilidad de los extractos obtenidos utilizando solventes de polaridad creciente. Posteriormente, se detectaron los metabolitos secundarios presentes en los extractos mediante la ejecución del tamizaje fitoquímico, donde se utilizaron diversos reactivos Shinoda, Dragendorff, Borntrager, entre otros. Se utilizó el método de cromatografía en capa fina, espectrofotometría UV/Vis y reactivos de desplazamiento para caracterizar las estructuras químicas de los flavonoides presentes en los extractos metanólicos de hojas de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Resultados. El extracto metanólico de hojas de Ruta chalepensis L. «ruda macho¼ presentó una mayor afinidad por solventes polares, mientras que en el extracto metanólico de hojas de Ruta chalepensis L. «ruda hembra¼ fue soluble en solventes medianamente polares. Se detectaron metabolitos secundarios tales como: taninos, alcaloides y flavonoides en ambos tipos. Por otro lado, se caracterizaron diez estructuras químicas tipo flavonoides a través del análisis de los espectros UV/Vis y utilizando reactivos de desplazamiento, de las cuales cinco corresponden a Ruta chalepensis L. «ruda macho¼ y las restantes a Ruta chalepensis L. «ruda hembra¼. Conclusiones. Se detectaron algunos metabolitos secundarios caracterizándose diez estructuras químicas de flavonoides en los extractos metanólicos de hojas de Ruta chalepensis L. «ruda macho¼ y «ruda hembra¼. Asimismo, la presencia de rutina en «ruda hembra¼ es la principal característica que la diferencia de «ruda macho¼.

Objective. To detect the secondary metabolites and characterize the chemical structures of the flavonoids in the methanolic extracts of leaves of the Ruta chalepensis L. "ruda macho" and "ruda hembra" types. Materials and methods. We elaborate the methanolic extracts of Ruta chalepensis L. leaves "ruda macho" and "ruda hembra". Then, the solubility test of the obtained extracts was carried out using solvents of increasing polarity. Subsequently, the secondary metabolites present in the extracts were detected by executing the phytochemical screening, various reagents were used: Shinoda, Dragendorff, Borntrager, among others. The method of thin layer chromatography, UV / Vis spectrophotometry and displacement reagents was used to characterize the chemical structures of the flavonoids in the methanolic extracts of leaves of the Ruta chalepensis L. "ruda macho" and "ruda hembra". Results. The methanolic extract of Ruta chalepensis L. "ruda macho" leaves showed a greater affinity for polar solvents, while the methanolic extract of Ruta chalepensis L. "ruda hembra" leaves was soluble in moderately polar solvents. Secondary metabolites were detected, such as: tannins, alkaloids and flavonoids in both types. On the other hand, ten flavonoidtype chemical structures were characterized through the analysis of UV / Vis spectra and using displacement reagent, of which five of them correspond to Ruta chalepensis L. "ruda macho" and the others to Ruta chalepensis L. "ruda hembra". Conclusions. Some secondary metabolites were detected and ten chemical structures of flavonoids were characterized in the methanolic extracts of Ruta chalepensis L. "ruda macho" and "ruda hembra" leaves. Likewise, the presence of rutina in "ruda hembra" is the main characteristic that differentiates it from "ruda macho".

Essential Oil Composition of Ruta graveolens L. Fruits and Hyssopus officinalis Subsp. aristatus (Godr.) Nyman Biomass as a Function of Hydrodistillation Time.

The aim of this study was to establish the kinetics regression models for yield and composition of Ruta graveolens fruit and Hyssopus officinalis subsp. aristatus aboveground biomass essential oil (EO), collected at different time intervals during the hydrodistillation process. The hypothesis was that collecting the EO fractions during specific time frames may result in EOs with dissimilar composition that may have differential use by the industry. Furthermore, we calculated the kinetics regression models for the composition of EO, isolated by hydrodistillation in a Clevenger-type apparatus and characterized by GC-MS and GC-FID analyses. The EO yield of R. graveolens fruits was 0.39% (relative area % of GC-FID chromatogram), with major constituents in the Control fraction (0-90 min) being 2-nonanone, 2-undecanone, and 2-undecanol, representing 65% of the total oil. The highest concentration of 2-nonanone (60%) was found in the 30-60 min oil fraction, the concentration of 2-undecanone (35%) was highest in the Control (0-90 min) fraction, and the concentration of eucalyptol (19%) was highest in the 5-10 min fraction. The EO yield of H. officinalis subsp. aristatus dried biomass was 1.12%. The major constituents in the Control fraction (0-90 min) of H. officinalis biomass were eucalyptol, α-pinene, sabinene, ß-pinene, and cis-3-pinanone, representing 86% of the total. Eucalyptol (58%) was the highest in the 0-5 min fraction. The highest ß-pinene (15%) and cis-3-pinanone (20%) contents were found in the 20-40 min fraction. The kinetics regression models that were developed for EO composition of R. graveolens were second-order polynominal, Michaelis-Menten, and Exponential decay, while for EO composition of H. officinalis subsp. aristatus biomass were Exponential decay and Power. The results from this study could benefit the EO industry.

8-Methoxycoumarin enhances melanogenesis via the MAPKase signaling pathway.

8-Methoxycoumarin (8-methoxy-chromen-2-one), isolated from R. graveolens L., is able to alleviate arthritis by inhibition of proinflammatory cytokines. However, its effects on melanogenesis have largely remained unreported. The present study examined the effects of 8-methoxycoumarin on melanogenesis in B16F10 murine cells, together with its effect on the mechanism of melanin synthesis. The cells were treated with different concentrations of 8-methoxycoumarin; α-MSH was used as the positive control. We found 8-methoxycoumarin to significantly increase the melanin content of the cells without exerting any cytotoxicity. In addition, it significantly upregulated the expression of tyrosinase and tyrosinase-related protein-1 and 2 via inducing the expression of microphthalmia-associated transcription factor. Furthermore, we demonstrated the involvement of mitogen-activated protein kinase (MAPK) pathway-mediated phosphorylation of p38 and c-Jun N-terminal kinase (JNK), and inhibition of phosphorylation of extracellular signal-regulated kinase (ERK) to be responsible for enhanced melanin production. Use of SB203580 (p38 inhibitor) and SP600125 (p-JNK inhibitor) corroborated these findings. Additionally, we investigated the effects of 8-methoxycoumarin on protein kinase B (AKT) phosphorylation and protein kinase A (PKA) signaling pathway (using H89, a PKA inhibitor). These results suggested that 8-methoxycoumarin increases melanogenesis via the MAPK signaling pathway. Based on these findings, we conclude that 8-methoxycoumarin could serve as a potential compound for treating hypopigmentation disorders. It could also serve as a promising chemical for hair depigmentation treatment in the cosmetic industry.

Colletotrichum Gloesporioides Inhibition In Situ by Chitosan-Ruta graveolens Essential Oil Coatings: Effect on Microbiological, Physicochemical, and Organoleptic Properties of Guava (Psidium guajava L.) during Room Temperature Storage.

Guava is a fruit appreciated worldwide for its high content of bioactive compounds. However, it is considered a highly perishable fruit, generally attacked by pathogenic species such as the fungi Colletotrichum gloeosporioides, which causes anthracnosis. To diminish the losses caused by pathogenic fungi, coatings of chitosan (CS) with Ruta graveolens essential oil (RGEO) in different concentrations (0.5, 1.0, 1.5% v/v) were applied in situ and their effects on the physical properties and microbiological quality of the guavas were studied. The CS+RGEO coated fruits exhibited better physicochemical behavior and lower microbiological decay as compared to the uncoated guavas, demonstrating the effectiveness of the coatings, especially those with 1.5% of RGEO content. All the fruits coated had greater acceptance and quality than the controls, being more those with essential oil incorporation. In situ investigation of C. gloesporioides infection of guavas demonstrated that the CS+RGEO coated guavas showed a high percentage of inhibition in the development of anthracnose lesions. In the present investigation, an alternative method has been proposed to extend the stability of the guavas fruit up to 12 days with application in the food industry.

Anticoccidial efficacy of Canary rue (Ruta pinnata) extracts against the caprine apicomplexan Eimeria ninakohlyakimovae.

Continuous use of anticoccidial treatments against Eimeria infections has resulted in the development of drug resistance. This study aimed to evaluate the anticoccidial efficacy of a methanolic extract derived from the endemic Canary rue (Ruta pinnata) plant of the Canary Islands, Spain, against Eimeria ninakohlyakimovae using in vitro assays. Freshly unsporulated oocysts were exposed to different concentrations of R. pinnata extract and thereafter evaluated for sporulation inhibition. Additionally, anticoccidial activity was examined by testing the viability of the E. ninakohlyakimovae sporozoites and their ability to infect bovine colonic epithelial cells after incubation with different concentrations of R. pinnata plant extract. The inhibition of oocyst sporulation by the extract was both time and concentration dependent, with certain combinations affording the same levels of sporulation inhibition as formaldehyde used as positive control (P < 0.001). Moreover, concentrations >0.1 mg/mL also affected not only the viability of the sporozoites but also their cell invasion capacity (P < 0.001). Altogether, these results show that methanolic fruit extracts from R. pinnata have important anticoccidial activity against oocysts and sporozoites of Eimeria. The potential efficacy of the extracts against other animal/human parasites remains to be elucidated, and further studies are needed to better understand its mode of action against coccidian parasites.

Ruta chalepensis L. (Rutaceae) leaf extract: chemical composition, antioxidant and hypoglicaemic activities.

Ruta chalepensis L. (Rutaceae) leaf extract was investigated for its chemical profile and antioxidant and hypoglycaemic properties. The antioxidant effects were investigated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ß-carotene bleaching, and metal chelating activity assays. The carbohydrate-hydrolysing enzymes inhibition assay was used to test the hypoglycaemic potential. R. chalepensis showed a high content of hesperidin and rutin with values of 591.9 and 266.7 mg/g dry extract, respectively. The extract exhibited a promising protection of lipid peroxidation (IC50 value of 16.9 µg/mL) and inhibited both α-amylase and α-glucosidase enzymes in a concentration-dependent manner. The highest activity was found against α-amylase (IC50 value of 69.0 µg/mL). The obtained results support the use of R. chalepensis leaves as healthy food ingredients.

Effect of Ruta chalepensis on Zinc, Lipid Profile and Antioxidant Levels in the Blood and Tissue of Streptozotocin-Induced Diabetes in Rats Fed Zinc-Deficient Diets.

OBJECTIVES: In diabetes, oxidative stress and lipid abnormalities are common and pronounced and represent important factors that are involved in the development of complications of diabetes. Zinc deficiency generally induces oxidative stress, but it is well known that the antioxidant Ruta chalepensis has an effective modulator role in oxidative stress in metabolic diseases. The aim of this study was to investigate the effect of R. chalepensis extract on blood biochemical parameters, tissue zinc status and antioxidant systems in rats with diabetes that were fed zinc-deficient diets. METHODS: We divided 28 male albino Wistar rats into 4 groups: 2 groups (1 group with diabetes, 1 group without diabetes) were fed zinc-sufficient diets, while the other 2 groups of rats with diabetes were fed zinc-deficient diets. One group was not treated, and the other was treated with the extract of R. chalepensis. After 3 weeks of dietary manipulation, the fasting animals were killed. RESULTS: The body-weight gains of the zinc-deficient animals with diabetes were lower than those of the zinc-adequate animals with diabetes. It was noticed also that inadequate dietary zinc intake increased the glucose, cholesterol, triglyceride, urea, uric acid, creatinine and lipid peroxidation levels. In addition, the zinc-deficient diet led to a decrease in zinc tissues (femur, liver, kidney), glutathione concentration and both glutathione peroxidase and glutathione-S-transferase activities. However, R. chalepensis treatment ameliorated all the previous parameters approximately to their normal levels. CONCLUSIONS: It seems that R. chalepensis supplementation is a potent factor in reducing the oxidative severity of zinc deficiency in experimental diabetes through its hypoglycemic and antioxidant actions.

Semisynthesis of (-)-Rutamarin Derivatives and Their Inhibitory Activity on Epstein-Barr Virus Lytic Replication.

(+)-Rutamarin inhibits EBV lytic DNA replication with an IC50 of 7.0 µM. (-)-Chalepin, a (-)-rutamarin derivative, was isolated from the whole plant of Ruta graveolens and used as a precursor of (-)-rutamarin. Altogether, 28 (-)-rutamarin derivatives were synthesized starting from (-)-chalepin. Of these, 16 compounds (2a-e, 3b-e, 3g, 4f, 4k, 4m-p) were found to be more potent against EBV lytic DNA replication than (-)-chalepin. Compounds 4m, 4n, and 4p exhibited IC50 values of 1.5, 0.32, and 0.83 µM and showed selectivity index values (SI) of 801, 211, and >120, respectively. Thus, compounds 4m, 4n, and 4p are considered promising leads for further laboratory investigation.