Ahmed Akrout - Academia.edu (original) (raw)

Papers by Ahmed Akrout

Les activites antiradicalaire. antioxydante, antibacterienne, antifongique et insecticide ont ete... more Les activites antiradicalaire. antioxydante, antibacterienne, antifongique et insecticide ont ete etudiees sur l'huile essentielle extraite par hydrodistillation a partir des feuilles d'Artemisia campestris L. (Asteracees) collectees au cours du mois d'aout dans quatre regions du sud-est de la Tunisie (Ben Guerdane, Beni kehdache, Djerba et Tataouine). Les resultats de cette etude ont montre que l'huile extraite a partir des echantillons collectes a Djerba, qui est relativement plus riche en composes oxygenes, est plus active que celles des autres provenances. Mais, ces activites restent moyennes a faibles par rapport a celles des composes de reference.

Institut des Régions Arides, 2012

South African Journal of Botany

Recent Research in Science and Technology, 2009

The present study was conducted to evaluate in vitro antibacterial and antiradical activities of ... more The present study was conducted to evaluate in vitro antibacterial and antiradical activities of essential oils extracted from air-dried leaves of Artemisia campestris Artemisia herba alba and Thymus capitatus growing wild in the southern of Tunisia. The principle compounds of Artemisia campestris oil were b -pinene (45.8%) and a -pinA¨ne (12.5%), the major constituents of Artemisia herba alba oil were I² -thujone (30.0%) and I± -thujone (25.7%) whereas the Thymus capitatus oil was mainly composed of  carvacrol (68.8%) and p-cymA¨ne (11.1%). The determination of the antiradical activity by DPPH method showed that Thymus capitatus oil exerted the highest activity with (0.15 µl/ml), followed by Artemisia herba alba (1.0 µl/ml) and Artemisia campestris (2.09 µl/ml). The screening of the antibacterial activity against seven bacteria using the disc diffusion method showed that Thymus capitatus oil strongly inhibited the growth of all bacteria studied (20 - 30 mm) except Pseudomonas ...

Journal of Medicinal Plants Research, 2011

The main objective of this study is to estimate the essential oil (EO) yield of Artemisia herba-a... more The main objective of this study is to estimate the essential oil (EO) yield of Artemisia herba-alba through biomass prediction. EO yield dependent in both the biomass production and its EO content, was determined from the recovered biomass after harvest of the upper half of plant tuft. The bio-volume parameter of twenty cultivated individuals of A. herba-alba were estimated four times during four years from height and diameters measurements. In the first harvest, after one year of establishment, several individuals appeared more productive than others. Thus, the canopy diameters of the plant tuft and the bio-volume recorded little significant relationship with the EO yield (R2 = 0.47 and 0.57, respectively). Harvesting the upper half of plant tuft, the spherical volume model developed by the regenerated material of the same individuals resulted in a high correlation between the EO yield and both the mean diameter (R2 = 0.61 to 0.62) and the bio-volume (R2 = 0.66 to 0.83). Key wor...

Essential oils (EO) from fresh and dry aerial parts of Pituranthos chloranthus (Benth.) Hook and ... more Essential oils (EO) from fresh and dry aerial parts of Pituranthos chloranthus (Benth.) Hook and Pituranthos tortuosus (Coss.) Maire were isolated by hydrodistillation and analyzed by GC/MS. The main constituents of the EO obtained from fresh herb of P. chloranthus were found to be α-pinene, sabinene, cis-ocimene and myrcene. In dry biomass, a significant increase of the content of some compounds such as α-phellandrene, △,3-carene and β-phellandrene characterized the oil. Minor changes in the chemical composition of the P. tortuosus EOs obtained from fresh or dry herbs and the major constituents were found to be sabinene and myrcene with equilibrate amounts of α-pinene, p-cymene, cis-ocimene, limonene, trans-β-ocimene, γ-terpinene and cis-verbenol. The paper disc diffusion method was used to evaluate the antibacterial activity and results showed an important inhibitory effect of oils obtained from fresh herb against most tested bacteria.

South African Journal of Botany

Abstract The aim of the study was to identify the best evaporation process that allows obtaining ... more Abstract The aim of the study was to identify the best evaporation process that allows obtaining a dry methanol extract of Moringa oleifera leaves rich on phytochemicals and showing a significant antioxidant activity. The liquid extract, prepared by the maceration of the leaves of M. oleifera into 70% aqueous methanol at room temperature, was evaporated using three solvent evaporation methods, these included evaporation in air at room temperature, with rotary evaporator and in an oven at 40 °C. Extracts were investigated for total polyphenols content (TPC), total flavonoids content (TFC), total condensed tannins content (TCTC), identification and quantification of some phenolic compounds by high-performance liquid chromatography coupled with photodiode array and mass spectrometry detection (HPLC-PDA-ESI/MS) and evaluated for antioxidant activity. The analysis of extracts by HPLC-PDA-ESI/MS showed that the dry extract obtained in dark after oven evaporation exhibited the highest amounts of 13 out of 19 phenolic compounds; quinic acid, gallic acid, 4-O-caffeoylquinic acid, caffeic acid, trans-ferulic acid, apigenin 7-O-glucoside, quercetrin, kaempferol, naringenin, apigenin, luteolin, cirsiliol and cirsilineol. In all extracts, quinic acid, 4-O-caffeoylquinic acid, rutin and quercitrin were present as the main compounds. The chemical composition and the antioxidant potential of the methanol extract are strongly affected by the method of evaporation of the solvent used. The highest values of TPC, TCTC and the antioxidant activity were provided by the extract obtained when the solvent was evaporated in oven. These results showed that the evaporation of the solvent in the oven at 40 °C was the best evaporation method to be used for the extraction of phytochemicals from the leaves of M. oleifera by cold maceration method using 70% aqueous methanol as solvent.

South African Journal of Botany

Abstract Ephedra alata is a medicinal plant with a long history of traditional use for its bronch... more Abstract Ephedra alata is a medicinal plant with a long history of traditional use for its bronchodilator, anti-asthmatic and cytotoxic effects. In this work, we develop a sensitive and validate LC-ESI/MS method to simultaneously identify and quantify 33 standard phenolic compounds in E. alata methanol (MeOH) crude extract and its derived fractions (Dichloromethane (DCM), Ethyl acetate (EAc), butanol (BuOH) and water). The antioxidant activities were tested using total antioxidant capacity (TAC), DPPH and FRAP assays. Results showed that the validated and reliable LC-ESI/MS method used unambiguously lead to identify and quantify 24 phenolic compounds most of them are present in all extracts. The major components detected were epicatechin, quinic acid, quercetin-3-o-rhamnoside, catechin, trans-cinnamic acid, naringin and trans-ferulic acid which their amounts vary according to the solvent used for the extraction. The DCM extract showed the highest activity when the TAC test was applied, the BuOH and EAc extracts displayed the highest activities with the DPPH test, whereas the DCM and EAc extracts exhibited the strongest activities with the FRAP method. This difference could be attributed to the nature and amounts of phenolic compounds of the extracts. These realized phytochemical and biological analyses could be helpful for the potential use of this plant in novel medicament products.

Preparative Biochemistry & Biotechnology

Cellular and Molecular Biology

Revue Des Regions Arides, 2007

The intraspecific chemical variability of essential oils (50 samples) isolated from the aerial pa... more The intraspecific chemical variability of essential oils (50 samples) isolated from the aerial parts of Artemisia herba-alba Asso growing wild in the arid zone of Southeastern Tunisia was investigated. Analysis by GC (RI) and GC/MS allowed the identification of 54 essential oil components. The main compounds were b-thujone and a-thujone, followed by 1,8-cineole, camphor, chrysanthenone, transsabinyl acetate, trans-pinocarveol, and borneol. Chemometric analysis (k-means clustering and PCA) led to the partitioning into three groups. The composition of two thirds of the samples was dominated by athujone or b-thujone. Therefore, it could be expected that wild plants of A. herba-alba randomly harvested in the area of Kirchaou and transplanted by local farmers for the cultivation in arid zones of Southern Tunisia produce an essential oil belonging to the a-thujone/b-thujone chemotype and containing also 1,8-cineole, camphor, and trans-sabinyl acetate at appreciable amounts. Introduction.-Artemisia herba-alba Asso (Asteraceae) is a silvery-green, perennial dwarf shrub growing in semiarid and arid climates. This species is known as desert wormwood in English, armoise blanche in French, and chih in Arab, and it is characteristic of the steppes and deserts of the Middle East (Egypt and Israel), North Africa (Tunisia, Algeria, and Morocco), and Southern Europe (Spain and Italy), extending into the Northwestern Himalaya [1]. In Tunisia, A. herba-alba is found from the mountains around Jebel Oust (Fahs) until the South of the country [2] [3]. The aerial parts of A. herba-alba are widely used in the traditional medicine to treat diabetes, bronchitis, diarrhea, and neuralgias [4-6]. The essential oil of this species is known for its disinfectant, anthelmintic, and antispasmodic therapeutic virtues [7]. Also the antibacterial and the antispasmodic activities of A. herba-alba essential oil from various chemotypes have been examined [8]. Moreover, the oil exhibited antileishmanial [9], spasmolytic [10], and antimutagenic activity against the carcinogen benzopyrene [11]. According to Lawrence [12] and Salido et al. [13], the aerial parts of A. herba-alba produce an essential oil characterized by a tremendous chemical variability. Various chemotypes have been reported in almost all the countries where A. herba-alba grows wild. Briefly, two types of oils could be distinguished: i) oils with a composition dominated by one major compound, i.e., camphor, a-thujone, b-thujone, chrysanthenone, chrysanthenyl acetate, or davanone, and ii) oils characterized by the occurrence at appreciable contents of two or more of these compounds.

Advances in Biological Chemistry, 2015

Journal of Essential Oil Bearing Plants, 2015

Revue Des Regions Arides, 1998

This study concerned four plants known for their use in traditional medicine in south-eastern Tun... more This study concerned four plants known for their use in traditional medicine in south-eastern Tunisia (Rosmarinus officinalis, Thymus capitatus, Artemisia herba-alba and Artemisia campestris) whose leaves are commonly consumed as tea. We aimed to determine the chemical composition and the antioxidant activity of these aqueous extracts prepared in accordance with the traditional method (50g/L of water).Because of the containment of flavonoids, polyphenols, tannins and saponins, all aqueous extracts have electrical conductivity (total salts) which ranged from 1.12 for the R. officinalis extract to 4.82 mS/cm for A. herba-alba. The last one is the richest species in potassium (575 mg/L) and especially sodium (400 mg/L). The content of total phenolics varied from 77.7 GAE mg/L of T. capitatus extract to 306.35 mg GAE/L of A. campestris extract.The antioxidant activities expressed as IC 50 values varied from 3.17 μL extract/mL of DPPH solution for the extract of A. campestris (the most a...

SUMMARY - "Study of essential oils of some pastoral plants from Matmata (Tunisia)" The ... more SUMMARY - "Study of essential oils of some pastoral plants from Matmata (Tunisia)" The essential oil of three plants (Artemisia herba-alba, Thymus capitatus and Juniperus phoenicea) collected from Matmata in the south east of Tunisia was isolated and investigated by GC and GC/SM. The yield of essential oil (ml/100 g) was 0.65% for Artemisia herba-alba, 2.75% for Thymus capitatus and 0.75% for Juniperus phoenicea. The Artemisia herba- alba was mainly composed of α-thujone (44%) and trans-acetate de sabinyle (17%). The principal component of Thymus capitatus essential oil was carvacrol (66%) while α-pinene (67.71%) constituted the major component of Juniperus phoenicea essential oil.

Eight annual species growing wild in the southern of Tunisia (Diplotaxis simplex, Chrysanthemum c... more Eight annual species growing wild in the southern of Tunisia (Diplotaxis simplex, Chrysanthemum coronarium, Matthiola longipetela, Erodium glaucophyllum, Reseda alba, Diplotaxis harra, Senecio gallicus and Papaver rhoeas) were evaluated for their mineral contents and phytochemical screening. The mineral analysis showed that calcium and potassium were the most concentrated minerals (1.21-3.60% and 0.36-3.20% respectively) followed by sodium (0.12-1.38%), magnesium (0.16-0.41%) and phosphorus (0.05-28%). The preliminary phytochemical screening revealed the absence of anthraquinones in all studied plants whereas alkaloids were only present in Papaver rhoeas. Senecio gallicus and Chrysanthemum coronarium were the only species that contained essential oils. All species were found to contain saponins, flavonoids and tannins with the exception of Erodium glaucophyllum and Papaver rhoeas. These results indicate that some of these species may be used as fodder plants for livestock due to the...

Les activites antiradicalaire. antioxydante, antibacterienne, antifongique et insecticide ont ete... more Les activites antiradicalaire. antioxydante, antibacterienne, antifongique et insecticide ont ete etudiees sur l'huile essentielle extraite par hydrodistillation a partir des feuilles d'Artemisia campestris L. (Asteracees) collectees au cours du mois d'aout dans quatre regions du sud-est de la Tunisie (Ben Guerdane, Beni kehdache, Djerba et Tataouine). Les resultats de cette etude ont montre que l'huile extraite a partir des echantillons collectes a Djerba, qui est relativement plus riche en composes oxygenes, est plus active que celles des autres provenances. Mais, ces activites restent moyennes a faibles par rapport a celles des composes de reference.

Institut des Régions Arides, 2012

South African Journal of Botany

Recent Research in Science and Technology, 2009

The present study was conducted to evaluate in vitro antibacterial and antiradical activities of ... more The present study was conducted to evaluate in vitro antibacterial and antiradical activities of essential oils extracted from air-dried leaves of Artemisia campestris Artemisia herba alba and Thymus capitatus growing wild in the southern of Tunisia. The principle compounds of Artemisia campestris oil were b -pinene (45.8%) and a -pinA¨ne (12.5%), the major constituents of Artemisia herba alba oil were I² -thujone (30.0%) and I± -thujone (25.7%) whereas the Thymus capitatus oil was mainly composed of  carvacrol (68.8%) and p-cymA¨ne (11.1%). The determination of the antiradical activity by DPPH method showed that Thymus capitatus oil exerted the highest activity with (0.15 µl/ml), followed by Artemisia herba alba (1.0 µl/ml) and Artemisia campestris (2.09 µl/ml). The screening of the antibacterial activity against seven bacteria using the disc diffusion method showed that Thymus capitatus oil strongly inhibited the growth of all bacteria studied (20 - 30 mm) except Pseudomonas ...

Journal of Medicinal Plants Research, 2011

The main objective of this study is to estimate the essential oil (EO) yield of Artemisia herba-a... more The main objective of this study is to estimate the essential oil (EO) yield of Artemisia herba-alba through biomass prediction. EO yield dependent in both the biomass production and its EO content, was determined from the recovered biomass after harvest of the upper half of plant tuft. The bio-volume parameter of twenty cultivated individuals of A. herba-alba were estimated four times during four years from height and diameters measurements. In the first harvest, after one year of establishment, several individuals appeared more productive than others. Thus, the canopy diameters of the plant tuft and the bio-volume recorded little significant relationship with the EO yield (R2 = 0.47 and 0.57, respectively). Harvesting the upper half of plant tuft, the spherical volume model developed by the regenerated material of the same individuals resulted in a high correlation between the EO yield and both the mean diameter (R2 = 0.61 to 0.62) and the bio-volume (R2 = 0.66 to 0.83). Key wor...

Essential oils (EO) from fresh and dry aerial parts of Pituranthos chloranthus (Benth.) Hook and ... more Essential oils (EO) from fresh and dry aerial parts of Pituranthos chloranthus (Benth.) Hook and Pituranthos tortuosus (Coss.) Maire were isolated by hydrodistillation and analyzed by GC/MS. The main constituents of the EO obtained from fresh herb of P. chloranthus were found to be α-pinene, sabinene, cis-ocimene and myrcene. In dry biomass, a significant increase of the content of some compounds such as α-phellandrene, △,3-carene and β-phellandrene characterized the oil. Minor changes in the chemical composition of the P. tortuosus EOs obtained from fresh or dry herbs and the major constituents were found to be sabinene and myrcene with equilibrate amounts of α-pinene, p-cymene, cis-ocimene, limonene, trans-β-ocimene, γ-terpinene and cis-verbenol. The paper disc diffusion method was used to evaluate the antibacterial activity and results showed an important inhibitory effect of oils obtained from fresh herb against most tested bacteria.

South African Journal of Botany

Abstract The aim of the study was to identify the best evaporation process that allows obtaining ... more Abstract The aim of the study was to identify the best evaporation process that allows obtaining a dry methanol extract of Moringa oleifera leaves rich on phytochemicals and showing a significant antioxidant activity. The liquid extract, prepared by the maceration of the leaves of M. oleifera into 70% aqueous methanol at room temperature, was evaporated using three solvent evaporation methods, these included evaporation in air at room temperature, with rotary evaporator and in an oven at 40 °C. Extracts were investigated for total polyphenols content (TPC), total flavonoids content (TFC), total condensed tannins content (TCTC), identification and quantification of some phenolic compounds by high-performance liquid chromatography coupled with photodiode array and mass spectrometry detection (HPLC-PDA-ESI/MS) and evaluated for antioxidant activity. The analysis of extracts by HPLC-PDA-ESI/MS showed that the dry extract obtained in dark after oven evaporation exhibited the highest amounts of 13 out of 19 phenolic compounds; quinic acid, gallic acid, 4-O-caffeoylquinic acid, caffeic acid, trans-ferulic acid, apigenin 7-O-glucoside, quercetrin, kaempferol, naringenin, apigenin, luteolin, cirsiliol and cirsilineol. In all extracts, quinic acid, 4-O-caffeoylquinic acid, rutin and quercitrin were present as the main compounds. The chemical composition and the antioxidant potential of the methanol extract are strongly affected by the method of evaporation of the solvent used. The highest values of TPC, TCTC and the antioxidant activity were provided by the extract obtained when the solvent was evaporated in oven. These results showed that the evaporation of the solvent in the oven at 40 °C was the best evaporation method to be used for the extraction of phytochemicals from the leaves of M. oleifera by cold maceration method using 70% aqueous methanol as solvent.

South African Journal of Botany

Abstract Ephedra alata is a medicinal plant with a long history of traditional use for its bronch... more Abstract Ephedra alata is a medicinal plant with a long history of traditional use for its bronchodilator, anti-asthmatic and cytotoxic effects. In this work, we develop a sensitive and validate LC-ESI/MS method to simultaneously identify and quantify 33 standard phenolic compounds in E. alata methanol (MeOH) crude extract and its derived fractions (Dichloromethane (DCM), Ethyl acetate (EAc), butanol (BuOH) and water). The antioxidant activities were tested using total antioxidant capacity (TAC), DPPH and FRAP assays. Results showed that the validated and reliable LC-ESI/MS method used unambiguously lead to identify and quantify 24 phenolic compounds most of them are present in all extracts. The major components detected were epicatechin, quinic acid, quercetin-3-o-rhamnoside, catechin, trans-cinnamic acid, naringin and trans-ferulic acid which their amounts vary according to the solvent used for the extraction. The DCM extract showed the highest activity when the TAC test was applied, the BuOH and EAc extracts displayed the highest activities with the DPPH test, whereas the DCM and EAc extracts exhibited the strongest activities with the FRAP method. This difference could be attributed to the nature and amounts of phenolic compounds of the extracts. These realized phytochemical and biological analyses could be helpful for the potential use of this plant in novel medicament products.

Preparative Biochemistry & Biotechnology

Cellular and Molecular Biology

Revue Des Regions Arides, 2007

The intraspecific chemical variability of essential oils (50 samples) isolated from the aerial pa... more The intraspecific chemical variability of essential oils (50 samples) isolated from the aerial parts of Artemisia herba-alba Asso growing wild in the arid zone of Southeastern Tunisia was investigated. Analysis by GC (RI) and GC/MS allowed the identification of 54 essential oil components. The main compounds were b-thujone and a-thujone, followed by 1,8-cineole, camphor, chrysanthenone, transsabinyl acetate, trans-pinocarveol, and borneol. Chemometric analysis (k-means clustering and PCA) led to the partitioning into three groups. The composition of two thirds of the samples was dominated by athujone or b-thujone. Therefore, it could be expected that wild plants of A. herba-alba randomly harvested in the area of Kirchaou and transplanted by local farmers for the cultivation in arid zones of Southern Tunisia produce an essential oil belonging to the a-thujone/b-thujone chemotype and containing also 1,8-cineole, camphor, and trans-sabinyl acetate at appreciable amounts. Introduction.-Artemisia herba-alba Asso (Asteraceae) is a silvery-green, perennial dwarf shrub growing in semiarid and arid climates. This species is known as desert wormwood in English, armoise blanche in French, and chih in Arab, and it is characteristic of the steppes and deserts of the Middle East (Egypt and Israel), North Africa (Tunisia, Algeria, and Morocco), and Southern Europe (Spain and Italy), extending into the Northwestern Himalaya [1]. In Tunisia, A. herba-alba is found from the mountains around Jebel Oust (Fahs) until the South of the country [2] [3]. The aerial parts of A. herba-alba are widely used in the traditional medicine to treat diabetes, bronchitis, diarrhea, and neuralgias [4-6]. The essential oil of this species is known for its disinfectant, anthelmintic, and antispasmodic therapeutic virtues [7]. Also the antibacterial and the antispasmodic activities of A. herba-alba essential oil from various chemotypes have been examined [8]. Moreover, the oil exhibited antileishmanial [9], spasmolytic [10], and antimutagenic activity against the carcinogen benzopyrene [11]. According to Lawrence [12] and Salido et al. [13], the aerial parts of A. herba-alba produce an essential oil characterized by a tremendous chemical variability. Various chemotypes have been reported in almost all the countries where A. herba-alba grows wild. Briefly, two types of oils could be distinguished: i) oils with a composition dominated by one major compound, i.e., camphor, a-thujone, b-thujone, chrysanthenone, chrysanthenyl acetate, or davanone, and ii) oils characterized by the occurrence at appreciable contents of two or more of these compounds.

Advances in Biological Chemistry, 2015

Journal of Essential Oil Bearing Plants, 2015

Revue Des Regions Arides, 1998

This study concerned four plants known for their use in traditional medicine in south-eastern Tun... more This study concerned four plants known for their use in traditional medicine in south-eastern Tunisia (Rosmarinus officinalis, Thymus capitatus, Artemisia herba-alba and Artemisia campestris) whose leaves are commonly consumed as tea. We aimed to determine the chemical composition and the antioxidant activity of these aqueous extracts prepared in accordance with the traditional method (50g/L of water).Because of the containment of flavonoids, polyphenols, tannins and saponins, all aqueous extracts have electrical conductivity (total salts) which ranged from 1.12 for the R. officinalis extract to 4.82 mS/cm for A. herba-alba. The last one is the richest species in potassium (575 mg/L) and especially sodium (400 mg/L). The content of total phenolics varied from 77.7 GAE mg/L of T. capitatus extract to 306.35 mg GAE/L of A. campestris extract.The antioxidant activities expressed as IC 50 values varied from 3.17 μL extract/mL of DPPH solution for the extract of A. campestris (the most a...

SUMMARY - "Study of essential oils of some pastoral plants from Matmata (Tunisia)" The ... more SUMMARY - "Study of essential oils of some pastoral plants from Matmata (Tunisia)" The essential oil of three plants (Artemisia herba-alba, Thymus capitatus and Juniperus phoenicea) collected from Matmata in the south east of Tunisia was isolated and investigated by GC and GC/SM. The yield of essential oil (ml/100 g) was 0.65% for Artemisia herba-alba, 2.75% for Thymus capitatus and 0.75% for Juniperus phoenicea. The Artemisia herba- alba was mainly composed of α-thujone (44%) and trans-acetate de sabinyle (17%). The principal component of Thymus capitatus essential oil was carvacrol (66%) while α-pinene (67.71%) constituted the major component of Juniperus phoenicea essential oil.

Eight annual species growing wild in the southern of Tunisia (Diplotaxis simplex, Chrysanthemum c... more Eight annual species growing wild in the southern of Tunisia (Diplotaxis simplex, Chrysanthemum coronarium, Matthiola longipetela, Erodium glaucophyllum, Reseda alba, Diplotaxis harra, Senecio gallicus and Papaver rhoeas) were evaluated for their mineral contents and phytochemical screening. The mineral analysis showed that calcium and potassium were the most concentrated minerals (1.21-3.60% and 0.36-3.20% respectively) followed by sodium (0.12-1.38%), magnesium (0.16-0.41%) and phosphorus (0.05-28%). The preliminary phytochemical screening revealed the absence of anthraquinones in all studied plants whereas alkaloids were only present in Papaver rhoeas. Senecio gallicus and Chrysanthemum coronarium were the only species that contained essential oils. All species were found to contain saponins, flavonoids and tannins with the exception of Erodium glaucophyllum and Papaver rhoeas. These results indicate that some of these species may be used as fodder plants for livestock due to the...