Large Field Screening for Resistance to Broomrape (Orobanche crenata Forsk.) in a Global Lentil Diversity Panel (GLDP) (Lens culinaris Medik.).

Broomrape (Orobanche crenata Forsk.) is a serious problem causing important losses to lentil (Lens culinaris Medik.) production and productivity in Mediterranean countries. Despite intensive breeding activities, no resistance sources against O. crenata have been identified so far. In this study, a Global Lentil Diversity Panel (GLDP) of 1315 genotypes including local populations, landraces, accessions, improved lines and released varieties were evaluated for their resistance to O. crenata under highly infested field conditions at ICARDA Marchouch research station, Morocco. The trial was conducted according to an augmented design with repeated susceptible checks. The best-performing genotypes were selected based on the correlations between Orobanche infestation parameters and agronomic performance. Results showed significant variation (p < 0.005) among the studied genotypes and between the tested genotypes and checks for BY, D2F, D2M, PH, EODW and NEO. Out of the 1315 tested genotypes, only (1%) showed high to moderate resistance levels to O. crenata. Most of these genotypes are improved lines originating from different breeding programs. the PCA analysis clustered all the tested genotypes into four different groups. Good resistance levels were recorded for the genotypes ILL7723, ILL 7982, ILL 6912, ILL 6415, ILL 9850, ILL 605, ILL 7915, ILL 1861 and ILL 9888 showing a parasitism index and grain yield ranging from 1.69 to 5.99 and 10.97 to 60.19 g m-2, respectively. Person's correlation showed significant negative correlations between agronomic traits and infestation parameters. Both the path and spatial analysis showed that the D2F, NEO, D2OE, SEV and parasitism index (PI) were the strongest driver traits that influenced the seed yield (SY).

Ultrasonic-assisted extraction for phenolic compounds and antioxidant activity of Moroccan Retama sphaerocarpa L. leaves: Simultaneous optimization by response surface methodology and characterization by HPLC/ESI-MS analysis.

This study was designed to optimize the ultrasound-assisted extraction of phenolic compounds and the antioxidant activity of Moroccan Retama sphaerocarpa extracts using response surface methodology (RSM). A central composite design has been conducted to investigate the effects of three factors: extraction period (X1), solvent concentration (X2), and solvent-to-material ratio (X3) on extraction yield, total phenolic content (TPC), flavonoids content (TFC), and antioxidant activity. The obtained results showed that the experimental values agreed with the predicted ones, confirming the capacity of the used model for optimizing the extraction conditions. The best extraction conditions for the simultaneous optimization were an extraction time of 38 min, a solvent concentration of 58%, and a solvent-to-material ratio of 30 mL/g. Under these conditions, the optimized values of yield, TPC, TFC, and DPPH-radical scavenging activity (DPPHIC50) were 18.91%, 154.09 mg GAE/g, 23.76 mg QE/g, and 122.47 µg/mL, respectively. The further HPLC/ESI-MS analysis of the obtained optimized extract revealed the presence of 14 phenolic compounds with piscidic acid, vitexin, and quinic acid as major compounds. These research findings indicate promising applications for efficiently extracting polyphenolic antioxidants, especially in the food industry.

Vasorelaxant Effect of Moroccan Cannabis sativa Threshing Residues on Rat Mesenteric Arterial Bed is Endothelium and Muscarinic Receptors Dependent.

Introduction: Ethanolic fraction of Moroccan Cannabis sativa threshing residues (EFCS) was evaluated for its vasorelaxant activity. The current work aims to identify the active metabolites in the ethanolic fraction of the EFCS and illustrate their mechanism of action. Methods: Free radical scavenging capacity of EFCS was assessed using DPPH method. The EFCS vasodilation activities in phenylephrine-precontracted isolated rat mesenteric arterial beds were investigated in presence of L-NAME (nitric oxide synthase inhibitor), indomethacin (cyclooxygenase inhibitor), potassium channel blockers (namely tetraetylamonium, barium chloride, and glibenclamide), and atropine. Nitric oxide vascular release was measured by electron paramagnetic resonance (EPR) using a spin trap in rat aortic rings. Results: EFCS induced dose-dependent vasorelaxation on mesenteric vascular bed. Incubation of the preparations with L-NAME, ODQ (a soluble guanylyl cyclase inhibitor), or potassium channel blockers reduced the fall of perfusion pressure caused by EFCS. Endothelial denudation or atropine abolished the EFCS's vasorelaxant effect, suggesting involvement of muscarinic receptors and endothelium-relaxing factors. The extract induced nitric oxide release in aortic rings in a similar manner as acetylcholine suggesting an effect of EFCS on the muscarinic receptor and the conductance arteries. Chemical investigation of EFCS identified potential active components namely apigenin and derivatives of luteolin skeleton and also additional components such as neophytadiene, squalene, and ß-sitosterol. In conclusion, the vasorelaxant effect of EFCS on rat mesenteric arterial bed, which is dependent of muscarinic receptor activation, nitric oxide, and EDHF, can account for potential therapeutic use against high blood pressure related cardiovascular diseases.

Ultra-High-Performance Liquid Chromatography with Photodiode Array and High-Resolution Time-of-Flight Mass Spectrometry Detectors (UHPLC-PDA-ESI-ToF/HRMS) for the Tentative Structural Characterization of Bioactive Compounds of Salvia verbenaca Extracts in Relation to Their Biological Activities.

BACKGROUND: Salvia verbenaca of the Lamiaceae family is a Mediterranean plant widely used in the Moroccan traditional folk medicine. The aim of this work was to explore the phytochemical composition of Salvia verbenaca extracts and its antioxidant activity. METHODS: Separation and identification of the major phytochemicals present in the two hexane and ethyl acetate explored extracts have been achieved through ultra-high-performance liquid chromatography separation technique coupled to photodiode array and high-resolution time-of-flight mass spectrometry detectors. Antioxidant activity of the obtained extracts was evaluated through DPPH• (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azinobis(3-ethylbenzothiazoline6-sulfonic acid) diammonium salt) assays and the obtained results were compared to TROLOX (6-hydroxy2,5,7,8-tetramethylchroman-2-carboxylic acid). RESULTS: Using the analysis technique indicated above, eighteen phytochemicals belonging to phenolic acids, phenolic diterpenes and flavonoids have been characterized on the basis of the obtained UV and mass spectroscopic properties and on the subsequent fragmentations. The antioxidant activity of the explored extracts showed potential scavenging activities compared to TROLOX. A detailed discussion of the attained results has been presented considering the activities observed of each extract. CONCLUSIONS: The research herein presented an analysis technique allowing to screen Salvia verbenaca phytochemicals. The explored plant could be considered as a source of functional phenolic compounds. These could be useful for further pharmacological studies such as new drugs design after clinic and its safety evaluation. It is thus hoped that the information presented here might prompt further studies that will possibly lead to development of therapeutic agents from this plant.

The effect of using diet supplementation based on argane (Argania spinosa) on fattening performance, carcass characteristics and fatty acid composition of lambs.

The current study was performed in order to evaluate the effect of using argane by-products (oil cake and pulp) as fattening diet of lambs. This was achieved through an experimental lamb fattening investigation using argan by-product as feeding source and the exploration of the fattened lamb performance, carcass characteristics and the chemical fatty acid composition of the quality produced meat. Twenty fattening Sardi lambs (26 ± 0.5 kg body weight and six months old) were divided into two (n = 10) homogenous experimental and control groups. Argane by-products (ABP) and traditional (CF) diets have been used for feeding the experimental and control groups respectively during a period of 75 days. The results obtained for both groups were used to determine the effect of feeding argane by-products on animal weight, average daily gain (ADG), carcass weight, muscle pH values, dressing %, dry matter, ash, ether extract, crude fiber, crude protein, and fatty acid composition. The obtained results showed that the experimental ABP group lambs had higher average daily gain and meat chemical, crude protein and ash and proportion of C18:0 than the control CF group lambs, while the control CF group had a higher feed conversion ratio, ether extract, mesenteric and perirenal fat. The results obtained for pH0 and pH24 were similar for both explored groups. The obtained results showed thus that the use of ABP as a diet to fattening lambs increased their performance and lean meat yield.

Structural characterization of bioactive compounds in Cotula cinerea extracts by ultra-high-performance liquid chromatography with photodiode array and high-resolution time-of-flight mass spectrometry detectors.

RATIONALE: Cotula cinerea of the Asteraceae family is a traditional Moroccan plant with various biological activities such as analgesic, cytotoxic and antioxidant effects which are often related to the presence of secondary metabolites. The present work aims to screen and identify the main phytochemicals compounds of Cotula cinerea extracts. METHODS: A method was developed that coupled a rapid and simple ultra-high-performance liquid chromatography system with both photodiode array and high-resolution time-of-flight mass spectrometry detectors (UHPLC-PDA/TOF-HRMS) for the identification of the main secondary metabolites of three investigated extracts (hexane, AcOEt and n-BuOH). RESULTS: A total of 30 phytocomponents pertaining to phenolic compounds and terpenoids have been detected, identified and quantified. Among these were previously reported free and conjugated coumaric and caffeic acids along with free and conjugated flavones and flavonols with kaempferol, quercetin, luteolin and apigenin aglycones. In addition, sulfated flavonoids were identified in the investigated extracts and are reported in this work for the first time in Cotula cinerea. The obtained results have been discussed in relation to the biological activities of the corresponding extracts. CONCLUSIONS: This study proposed a practical strategy for rapidly screening and identifying secondary metabolites of Cotula cinerea and provided new information on the phytochemicals of this Saharan plant. This work has therefore provided useful results for further pharmacological studies and the design of new drugs based on this species and will facilitate the utilization of Cotula cinerea in the clinic and its safety evaluation. It is also hoped that the information presented here might stimulate further studies that will possibly lead to development of therapeutic agents from this plant.

The effect of including argane by-products (meal and pulp) in dairy ewe diet on milk quality and production, and lamb growth performance.

The objective of this work was to study the effect of the incorporation of argane by-products (meal and pulp) in ewe's diet on the production and quality of milk and the performance of lambs in the lactation phase. Twenty ewes were divided into two equal groups. The first one has been fed with argane by-product diet (AD), and the second with a control diet (CD). The results showed an improvement in milk production with an average of 26.3% for AD compared with the CD group. AD group lambs showed a significant difference (p < 0.05) compared with the CD group during lactation phase. The ash and protein levels were not affected by treatments; however, fat, lactose, density, and fusion point content show more fluctuations for both treatments (p < 0.05). The physicochemical parameters of sheep milk showed high (p < 0.01) to very high (p < 0.001) significant change, along the lactation weeks. Except the Zn which showed no significant difference (p > 0.05), the mineral composition of both milks (Na, K, Ca, Mg, P, and Fe) was significant (p < 0.05). It is concluded that, in terms of performances, argane by-products could replace the conventional diet for ewes without substantial detrimental effects. Therefore, argane by-product could be used as a cost-effective feed for sheep in dry areas.

Cytotoxicological Investigation of the Essential Oil and the Extracts of Cotula cinerea and Salvia verbenaca from Morocco.

The objective of this work was to investigate the cytotoxicological effect of the extracts (hexane, ethyl acetate, and n-butanol) of Cotula cinerea and Salvia verbenaca in addition to the essential oil of Cotula cinerea. These plants are widely used in the Moroccan traditional folk medicine. The cytotoxic effect was explored against two cancer cell lines, Vero and RD, using the colorimetric MTT assay. The obtained results showed that the cytotoxicity differed according to the used extract with an efficient effect of Cotula cinerea extracts compared to Salvia verbenaca. A potent cytotoxicity was thus observed for the Cotula cinerea hexane extract which inhibited the growth of RD cell line at the lowest IC50 value (57.21±3.43 µg/mL). This was followed by the ethyl acetate extract and the essential oil with moderate effects against RD cell line and showed IC50 values of 187.52±6.27 µg/mL and 173.05±4.46 µg/mL, respectively. On the other hand, different results were obtained and Cotula cinerea essential oil was the most cytotoxic with the lowest IC50 value (72.72±2.18 µg/mL) against Vero cell line. In the same conditions, higher concentrations were needed in the case of Salvia verbenaca extracts. The results of this study showed thus that Cotula cinerea essential oil and hexane extract showed significant cytotoxic effects against RD and Vero cell lines, respectively, and could be considered as novel source of antitumor agents. This study is expected to be beneficial for clinical and traditional applications for Cotula cinerea as a remedy against cancer and opens new perspectives for further investigations on other types of cancer cell lines.

Antimicrobial effect of the Tunisian Nana variety Punica granatum L. extracts against Salmonella enterica (serovars Kentucky and Enteritidis) isolated from chicken meat and phenolic composition of its peel extract.

Punica granatum L. is widely recognized for its potency against a broad spectrum of bacterial pathogens. The purpose of this study was to explore the inhibitory and the bactericidal activities of Punica granatum against Salmonella strains. The effect of extracts obtained from different parts (peels, seeds, juice and flowers) of pomegranate and using different solvents against Salmonella enterica serovars Kentucky and Enteritidis isolated from chicken meat was thus investigated. Salmonella strains were identified with the standard API-20E system and confirmed by real time PCR. The obtained results showed that the highest antibacterial activity against Salmonella strains was observed with the peels ethanolic extract giving MIC values ranging from 10.75 to 12.5mg/mL. The ethanolic extract of P. granatum Nana peels at 0.8 and 1.6mg/g significantly inhibited the growth of Salmonella Kentucky in chicken meat stored at 4°C. The phenolic composition of the ethanolic peel extract was explored by HPLC coupled to both DAD and ESI/TOF-MS detections. The obtained results allowed the detection of 21 phytochemical compounds among which various phenolic compounds have been identified on the basis of their UV and MS spectra as well as with literature data. Among the detected compounds, anthocyanins, ellagitannins, ellagic acid derivatives and flavanols were further characterized through MS-MS analysis. Our results showed thus that the Tunisian variety Nana pomegranate constitutes a good source of bioactive compounds with potent antimicrobial activity on the growth of Salmonella strains suggesting that the studied pomegranate cultivar could be a natural remedy to minimize the emergence of Salmonella enterica strains which is often involved in food borne illness.

Characterization of flavonoid glycosides from fenugreek (Trigonella foenum-graecum) crude seeds by HPLC-DAD-ESI/MS analysis.

Fenugreek (Trigonella foenum-graecum) is a medicinal plant which is widely used for its pharmacological properties. In this study the phenolic composition of fenugreek crude seeds originating from Morocco has been investigated. Extraction was performed from defatted seeds by a hydromethanolic solution using an Accelerated Solvent Extractor. HPLC technique coupled to negative ion electrospray ionization mass spectrometry and diode array detection was employed to identify the polyphenol in the obtained extract. The obtained results allowed the detection of 32 phenolic compounds among which various flavonoid glycosides and phenolic acids have been tentatively identified on the basis of their UV and MS spectra, and comparisons with standards when available, as well as with literature data. A systematic study of the obtained MS spectra and the observed fragmentation showed that most of the identified compounds were acylated and non-acylated flavonoids with apigenin, luteolin and kaempferol as aglycons. Hydroxycinnamic acids mostly dominated by caffeic acid derivatives were also detected. The quantitative analysis of the identified compounds showed that the phenolic composition of the studied crude fenugreek seeds was predominantly acylated and non-acylated flavone derivatives with apigenin as the main aglycon.

Gas-phase fragmentation study of novel synthetic 1,5-benzodiazepine derivatives using electrospray ionization tandem mass spectrometry.

The fragmentation patterns of a series of three novel synthesized 3-hydroxy-4-phenyl-tetrahydro-1,5-benzodiazepin-2-ones (1-3), possessing the same backbone structure, were investigated using electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) techniques. A simple methodology, based on the use of ESI (positive ion mode) and by increasing the declustering potential in the atmospheric pressure/vacuum interface, collision-induced dissociation (CID), was used to enhance the formation of the fragment ions. In general, the novel synthetic 1,5-benzodiazepine derivatives afforded, in the gas phase, both protonated and sodiated molecules. This led to the confirmation of the molecular masses and chemical structures of the studied compounds. Exact accurate masses were measured using a high-resolution ESI-quadrupole orthogonal time-of-flight (QqToF)-MS/MS hybrid mass spectrometer instrument. The breakdown routes of the protonated molecules were rationalized by conducting low-energy collision CID-MS/MS analyses (product ion- and precursor ion scans) using a conventional quadrupole-hexapole-quadrupole (QhQ) tandem mass spectrometer. All the observed major fragmentations for the 1,5-benzodiazepines occurred in the saturated seven-membered ring containing the nitrogen atoms. These formed a multitude of product ions by different breakdown routes. All the major fragmentations involved cleavages of the N-1-C-2 and C-3-C-4 bonds. These occurred with concomitant eliminations of glyoxal, benzene and ethyl formate, forming the product ion at m/z 119, which was observed in all the studied compounds. In addition, an unique simultaneous CID-MS/MS fragmentation was noticed for the 1,5-benzodiazepines 1 and 3, which occurred by a pathway dictated by the substituent located on the N-1-position. It was evident that the aromatic ring portion of the 1,5-benzodiazepines was resistant to CID-MS/MS fragmentation. Re-confirmation of the various geneses of the product ions was achieved by conducting a series of precursor ion scans. ESI-MS and CID-MS/MS analyses have thus proven to be a specific and very sensitive method for the structural identification of these novel 1,5-benzodiazepine derivatives.

New compounds obtained by evolution and oxidation of malvidin 3-O-glucoside in ethanolic medium.

Two new colorless phenolic compounds were formed from malvidin 3- O-glucoside incubated in an ethanolic solution. Their structures were characterized by means of one- and two-dimensional NMR analysis and through electrospray ionization-mass spectrometry. As compared to the structure of the initial anthocyanin skeleton, the first new compound showed the presence of two fused five-membered rings replacing the pyran ring and of a carbonyl function on the 2-position. The first five-membered ring was shown to result from the formation of a new linkage between the B ring 6'-position and the C ring 4-position, while the second was a dihydro furan ring with an oxygenated ether linkage between the 8a-position and the 3-position. The second isolated compound was shown to have similar structure with an ethyl ether moiety in the 3-position instead of the glucose moiety. A mechanism explaining the formation of the isolated compounds involving the passage through the chalcone form of the anthocyanin and an oxidation process is proposed.

Flavonoids: hemisynthesis, reactivity, characterization and free radical scavenging activity.

Phenolic compounds form one of the main classes of secondary metabolites. They display a large range of structures and they are responsible for the major organoleptic characteristics of plant-derived-foods and beverages, particularly color and taste properties and they also contribute to the nutritional qualities of fruits and vegetables. Phenolic compounds are also highly unstable compounds which undergo numerous enzymatic and chemical reactions during postharvest food storage and processing thus adding to the complexity of plant polyphenol composition. Among these compounds flavonoids constitute one of the most ubiquitous groups of all plant phenolics. Owing to their importance in food organoleptic properties and in human health, a better understanding of their structures, their reactivity and chemical properties in addition to the mechanisms generating them appears essential to predict and control food quality. The purpose of this work is an overview of our findings concerning the hemisynthesis, the reactivity and the enzymatic oxidation of some flavonoids and shed light on the mechanisms involved in some of these processes and the structures of the resulting products. The free radical scavenging activity of some of the synthesized compounds is also presented and a structure-activity relationship is discussed. The first part of this review concerns the synthesis and structural characterization of modified monomeric flavanols. The use of these compounds as precursor for the preparation of natural and modified dimeric procyanidin derivatives was then explored through different coupling reactions. The full characterization of the synthesized compounds was achieved by concerted use of NMR and ESI-MS techniques. The free radical scavenging activity of some of the synthesized compounds was investigated. The second part of this review concerns the enzymatic oxidation of several flavonols by Trametes versicolor laccase. Most of the major oxidation products have been isolated as pure compounds and their structures unambiguously established through spectroscopic methods. Correlation between the structure of the oxidation product and the substitution pattern of the starting materials allows mechanistic features of this transformation to be elucidated.