Chemical Characterization and Multidirectional Biological Effects of Different Solvent Extracts of Arum elongatum: in Vitro and in Silico Approaches

Arum elongatum (Araceae) is widely used traditionally for the treatment of abdominal pain, arterial hypertension, diabetes mellitus, rheumatism and hemorrhoids. This study investigated the antioxidant properties, individual phenolic compounds, total phenolic and total flavonoid contents (HPLC/MS analysis), reducing power and metal chelating effects of four extracts obtained from A. elongatum (ethyl acetate (EA), methanol (MeOH), methanol/water (MeOH/water) and infusion). The inhibitory activity of the extracts were also determined against acetylcholinesterase, butyrylcholinesterase, tyrosinase, amylase and glucosidase enzymes. The MeOH/water extracts contained the highest amount of phenolic contents (28.85 mg GAE/g) while the highest total flavonoid content was obtained with MeOH extract (36.77 mg RE/g). MeOH/water demonstrated highest antioxidant activity against DPPH⋅ radical at 38.90 mg Trolox equivalent per gram. The infusion extract was the most active against ABTS⁺⋅ (133.08 mg TE/g). MeOH/water extract showed the highest reducing abilities with the CUPRAC value of 102.22 mg TE/g and the FRAP value of 68.50 mg TE/g. A strong metal chelating effect was observed with MeOH/water extract (35.72 mg EDTAE/g). The PBD values of the extracts ranged from 1.01 to 2.17 mmol TE/g. EA extract displayed the highest inhibitory activity against AChE (2.32 mg GALAE/g), BChE (3.80 mg GALAE/g), α-amylase (0.56 mmol ACAE/g) and α-glucosidase (9.16 mmol ACAE/g) enzymes. Infusion extract was the most active against tyrosinase enzyme with a value of 83.33 mg KAE/g. A total of 28 compounds were identified from the different extracts. The compounds present in the highest concentration were chlorogenic acids, 4-hydroxybenzoic acid, caffeic acid, p-coumaric acid, ferulic acid, isoquercitrin, delphindin 3,5-diglucoside, kaempferol-3-glucoside and hyperoside. The biological activities of A. elongatum extracts could be due to the presence of compounds such as gallic acid, chlorogenic acids, ellagic acid, epicatechin, catechin, kaempferol, 4-hydroxybenzoic acid, caffeic acid, p-coumaric acid, ferulic acid, quercetin, isoquercitrin, and hyperoside. Extracts of A. elongatum showed promising biological activities which warrants further investigations in an endeavor to develop biopharmaceuticals.     

A substance inducing teliospore production in wheat leaf rust,Puccinia recondita f. sp.tritici

A substance inducing teliospore production inPuccinia racondita f. sp.tritici was found in water and methanol extracts of wheat leaves with telia of the wheat leaf rust just before harvest time. Methanol (MeOH) and water extracts from uninfected wheat leaves also showed telia-inducing activity. However, the MeOH and water extracts from wheat leaves covered with telia showed much stronger activity than those from uninfected wheat leaves. We obtained a fraction (0.2 mg) showing activity at 2 ng/ml by purification of the water extract.     

UHPLC/HR‐ESI‐MS/MS Profiling of Phenolics from Tunisian Lycium arabicum Boiss. Antioxidant and Anti‐lipase Activities’ Evaluation

This study was performed in the aim to evaluate nine different extracts from Tunisian Lycium arabicum for their total phenolic and total flavonoid contents, phytochemical analyses as well as their antioxidant and anti‐lipase activities. The in vitro antioxidant property was investigated using three complementary methods (DPPH, ferric reducing antioxidant power (FRAP), and β‐carotene‐linoleic acid bleaching assays) while anti‐lipase activity was evaluated using 4‐methylumbelliferyl oleate method. From all of the tested extracts the most potent found to be the polar MeOH extracts especially those of stems and leaves. In order to investigate the chemical composition of these extracts and possible correlation of their constituents with the observed activities, an UHPLC/HR‐ESI‐MS/MS analysis was performed. Several compounds belonging to different chemical classes were tentatively identified such as rutin and kampferol rutinoside, the major constituents of the leaves, and N‐caffeoyltyramine, lyciumide A, N‐dihydrocaffeoyltyramine as well as fatty acids: trihydroxyoctadecadienoic acid and hydroxyoctadecadienoic acid isomers were detected abundantly in the stems. These results showed that the MeOH extracts of stems and leaves of L. arabicum can be considered as a potential source of biological active compounds.     

Cytotoxicity of extracts from Dolsan leaf mustard Kimchi treated with lactic acid bacteria on lung and gastric cancer cells

The cytotoxicity of extracts from Dolsan leaf mustard Kimchi (DLMK) treated with lactic acid bacteria on A 549 human lung cancer cells and SNU-601 human gastric cancer cells were investigated. Leuconostoc mesenteroides, Leu. Gelidum, and Weissella kimchii previously isolated from properly ripened DLMK were inoculated to DLMK as a starter (1 × 10⁸ CFU/mL). The DLMK was then fractionated by various extracting solvents. The cytotoxicity of MeOH extracts from DLMK on A 549 and SNU-601 cancer cells was found to occur in a dose-dependent manner. Although the cytotoxicity of the MeOH extracts was found to be approximately 20 to 30% at concentrations of 250 μg/mL by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide) assay, cytotoxicity of chloroform soluble fraction of DLMK treated with W. kimchii showed about 80 to 90%. Consequently, the growth of cancer cells was inhibited significantly in medium containing DLMK extracts. In addition, significant morphological changes such as cell condensation, cell fragmentation, and alterations in the size and shape of the cells were observed in cells grown in medium that contained the DLMK extracts. Taken together, these results suggest that inhibition of the proliferation of cancer cells by apoptosis was induced by DLMK extracts.     

Antioxidant activity of the chemical constituents from the flower buds of <Emphasis Type="Italic">Magnolia denudata</Emphasis>

The scavenging activity of the flower buds of Magnolia denudata Desrousseaux on reactive oxygen species (ROS) was evaluated using 2′,7′-dichlorofluorescin diacetate (DCFH-DA) in HT 1080 cells. Methanol (MeOH) and dichloromethane (CH₂Cl₂) extracts inhibited dose-dependently generation of ROS in the cellular system. MeOH and CH₂Cl₂ extracts were combined and fractionated with n-hexane, 85% aqueous MeOH, and n-butanol (n-BuOH). Both n-hexane-soluble and 85% aqueous-soluble fractions showing strong radical-scavenging activity in the cellular system were further separated by diverse chromatographic methods to give five known lignans (1–5). All these compounds exhibited significant radical-scavenging effect on intracellular ROS in a dose-dependent manner. Their scavenging activity on various reactive oxygen species (ROS) was also evaluated using electron spin resonance (ESR) spin-trap techniques.     

Teliospore-inducing activity for wheat leaf rust in the extracts of various host plant leaves infected with telia of rust fungi

MeOH and water extracts were obtained from 16 species of infected leaves with rust fungi belonging to 18 species in 6 families: Pucciniaceae, Melampsoraceae, Coleosporiaceae, Pileolariaceae, Phragmidiaceae, and Phakopsoraceae. All the extracts of rust-infected plants with telia showed the teliospore-inducing activity for wheat leaf rust (Puccinia recondita f. sp. tritici).     

Leishmania amazonensis: Partial purification and study of the biochemical properties of the telomerase reverse transcriptase activity from promastigote-stage

Telomeres are protein–DNA complexes that protect chromosome ends from degradation and fusion. In Leishmania spp., telomeric DNA comprises a conserved TTAGGG repeat and is maintained by telomerase. Telomerase is a multisubunit enzymatic complex that ensures the complete DNA replication by adding new telomeric repeats to the G-rich strand. In this report we aimed to purify and study the biochemical properties of Leishmani amazonensis telomerase. In a first trial we used affinity chromatography with antisense 2′-O-methyl oligonucleotide without success since the Leishmania telomerase, similarly to Trypanosoma cruzi enzyme, was not eluted by competition, but instead, it remained bound to the column. Partially purified L. amazonensis telomerase activity was achieved by fractionation of extracts on complementary ion exchange and Heparin columns. Further purification of these fractions on a G-rich telomeric DNA affinity chromatography enriched for telomerase activity. The knowledge of telomerase characteristics in Leishmania could help to develop new strategies to overcome leishmaniasis.     

Barks Essential Oil,Secondary Metabolites and Biological Activities of Four Organs of Tunisian Calligonum azel Maire

This study is the first to investigate the chemical composition of barks essential oil (EO), secondary metabolites and biological activities of the MeOH and infusions extracts of seeds, leaves, barks and roots of Calligonum azel Maire (Polygonaceae) harvested from Tunisian desert. The gas chromatography/mass spectrometry (GC/MS) results showed the presence of fifty‐four compounds in barks EO. The major components were: viridiflorol (14.6%), α‐eudesmol (8.65%), trans‐caryophyllene (6.72%), elemol (6.63%), β‐eudesmol (6.21%). The obtained results showed that C. azel is a very rich plant in secondary metabolites. High contents in polyphenols, flavonoids and tannins were observed in both extracts of all studied organs. Significant differences were found between both extracts of the four organs. Thus, polyphenols and tannins were more abundant in leaves infusion extract, while, flavonoids showed a high level in barks extract. The antioxidant activity data demonstrated that all extracts showed strong antioxidant and radical scavenging activities. The MeOH extracts presented potential for antibacterial and antifungal activities against all tested microorganisms. The inhibition zones diameters and minimal inhibitrice concentration values were in the range of 9 – 15 mm and 2.5 – 20 μg/ml, respectively. This study demonstrated that C. azel can be regarded as an excellent plant source for natural antimicrobial agents.     

LC-ESI-MS/MS Chemical Characterization,Antioxidant and Antidiabetic Properties of Propolis Extracted with Organic Solvents from Eastern Anatolia Region

Geographic conditions (altitude, climate, and local flora) lead to significant differences in the chemical composition of propolis. Therefore, more research is needed for propolis in different geographical regions. So, the aim of this study was to evaluate the phenolic profile, total phenolic content, antioxidant, and antidiabetic properties of Pülümür propolis from Turkey. Methanol (MeOH), chloroform (CHCl₃), and hexane extracts of propolis were analyzed. LC-ESI-MS/MS analysis of the extracts showed that the most abundant phenolic compound is caffeic acid in the MeOH extract (2943.12±11.12 μg phenolics/g extract), while on the other hand, CHCl₃ extract had the highest total phenolic content (125.75±1.02 mg GAE/g extract). Antioxidant activity was measured using ABTS and DPPH assays, whereas CHCl₃ extract (IC₅₀=6.35±0.11 and 28.84±0.10 μg/mL, respectively) and MeOH extracts (IC₅₀=5.04±0.07 and 28.80±0.09 μg/mL, respectively) showed relatively high antioxidant activity. The MeOH extract showed better antidiabetic activity than the standard compound, acarbose (IC₅₀=0.544 and 0.805 mg/mL, respectively).     

Antioxidant,Anti-Skin-Aging,Anti-Inflammatory,and Anti-Acetylcholinesterase Activities of Rourea oligophlebia Extracts

The objective of this study was to evaluate the antioxidant, anti-skin-aging, anti-inflammatory, and anti-acetylcholinesterase activities of the hexane (n-hex), AcOEt, BuOH, MeOH, and aqueous extracts from R. oligophlebia roots. The total phenolic and flavonoid contents (TPC and TFC) were determined using Folin-Ciocalteu and AlCl₃ colorimetric assays. The antioxidant capacity was examined by reducing power (RP), ferric reducing antioxidant power (FRAP), ABTS⋅⁺, and DPPH⋅⁺ radical cation assays. All extracts potentially exhibited antioxidant activity with IC₅₀ values ranging from 2.93 to 5.73 μg/mL for ABTS⋅⁺ and from 5.69 to 7.65 μg/mL for DPPH⋅⁺ except the n-hex extract. The BuOH, MeOH, and aqueous extract possess promising anti-skin-aging activities, as observed by an attenuation of UV-A toxicity on human keratinocytes. We proposed that these anti-skin-aging properties are possibly due to direct scavenging activity against reactive oxygen species and upregulate cellular antioxidant machinery. Moreover, we found that the antioxidant capacity was well correlated with anti-inflammatory capacity against nitric oxide (NO) production in terms of the n-hex, AcOEt, and BuOH extracts with IC₅₀ values from 23.21 to 47.1 μg/mL. In contrast, these activities were found to be poorly correlated with AchE activity. To the best of our knowledge, this is the first report of the antioxidant, anti-skin-aging, anti-inflammatory, and anti-acetylcholinesterase activities of the extracts of R. oligophlebia roots. These findings indicated that this species could be a potential source of natural antioxidant, anti-aging, and anti-inflammatory agents. Consequently, it may be suggested as a medicinal plant that prevents diseases related to oxidative stress and inflammatory responses.     

Phytochemical and Biological Activities of Pseudocalymma elegans: A False Garlic

Evaluation of phytochemical constituents and antioxidant and antimicrobial activities of hexane (PELH), dichloromethane (PELDCM), ethyl acetate (PELEA), and MeOH (PELM) extracts of young leaves of Pseudocalymma elegans have been carried out. Moreover, extracts have also been explored for the presence of sulphur containing compounds, 1,2‐dithiolane ( 33 ), diallyl disulfide ( 35 ), 3‐vinyl‐1,2‐dithiacyclohex‐5‐ene ( 37 ), and diallyl trisulfide ( 38 ) responsible for the garlic like smell of P. elegans. All the extracts were found to be antioxidant and showed potent inhibition with IC₅₀ values of 0.168 ± 0.001, 0.128 ± 0.002, 0.221 ± 0.011, and 0.054 ± 0.001, respectively, as compared to standard drugs ascorbic acid (AA) and butylated hydroxytoluene (BHT). The ethyl acetate extract (PELE) showed excellent activities against few Gram‐positive and Gram‐negative bacteria and some fungi as compared with standard drug ceftriaxone (3rd generation cephalosporin) and nystatin, respectively. Chemical constituents of hexane, dichloromethane, and ethyl acetate extracts were identified by gas chromatography‐mass spectrometry and mass spectral library search. Over all 55 chemical constituents were first time identified from the leaves which included branched and n‐hydrocarbons, fatty acids, fatty acid methyl esters, fatty alcohols, terpenes, alkaloid, vitamins, glycosides, aromatic compounds, and sulfur containing compounds. Two known chemical constituents, ursolic acid ( 1 ) and β‐amyrin ( 2 ), were also purified for the first time from the MeOH extract. To elucidate the structures of these compounds, UV, IR, EI‐MS, ¹H‐ and ¹³C‐NMR spectroscopy were used.     

Evaluation of Antioxidant,Anticholinesterase, and Antidiabetic Potential of Dry Leaves and Stems in Tamarix aphylla Growing Wild in Tunisia

Tamarix aphylla (L.) Karst . has a wide geographic distribution and was employed in traditional medicine as astringent, anti‐rheumatic and to treat fever. T. aphylla leaves and stems extracts were studied from both chemical and biological points of view to assess the antidiabetic, anticholinesterase and antioxidant potential of this species. The HPLC/Diode Array Detector (DAD) analysis showed the presence of 14 phenolic compounds (gallic, caffeic, p‐coumaric, ferulic and ellagic acids, kaempferol, quercetin, quercetin 3‐O‐galactoside and six flavonol derivatives). This is the first study reporting a comparative study of the biological activities of different extracts from T. aphylla. High activities were obtained against DPPH radical, superoxide anion radical () and nitric oxide radical (^?NO) in a concentration‐dependent manner, the most active extracts being the polar ones. T. aphylla also showed moderate protective effects against acetylcholinesterase, but no effects were observed against butyrylcholinesterase. Against α‐glucosidase the MeOH extracts displayed IC₅₀ values from 8.41 to 24.81 μg/ml.     

Telomeric length and telomerase activity vary with age in peripheral blood cells obtained from normal individuals

The telomerase activity and length of telomeres of peripheral blood mononuclear cells obtained from 124 healthy individuals aged 4–95years was measured. Telomerase activity level was semiquantitatively assessed by a fluorescent-telomeric repeat amplification protocol (fluorescent-TRAP) using an internal telomerase assay standard, fluorescent primers and an automated laser fluorescent DNA sequencer. Telomeric length, measured by assay of terminal restriction fragments (TRFs), was determined in HinfI-digested DNA by Southern blot analysis using a (TTAGGG)₄ probe. TRF length was determined in 80 individuals and age-related progressive reduction of size was observed. TRF length in peripheral blood mononuclear cells obtained from normal individuals (aged 4–39years) decreased by approximately 84bp per year, while in individuals aged ≥40years it decreased by 41bp per year. In contrast, telomerase activity showed an apparent biphasic pattern with aging. Individuals aged 4–39years showed a progressive decrease in telomerase activity, whereas 65% of those aged ≥40years showed relatively stable but very low telomerase activity, and the remaining individuals aged ≥40years had no detectable telomerase activity. These data obtained from normal individuals might in the future be of value to help risk stratify and manage the care of patients with leukemia. Received: 18 August 1997 / Accepted: 10 December 1997     

Tulip chlorotic blotch virus, a second potyvirus causing tulip flower break

Tulip chlorotic blotch virus (TCBV), an apparently undescribed potyvirus found in field grown tulips in Australia, causes symptoms in tulip leaves and flowers identical to those induced by tulip breaking virus (TBV). TCBV was transmitted mechanically to 14 of 34 species in four of 13 families. Nicotiana clevelandii is a suitable propagation host and Chenopodium amaranticolor a local-lesion assay host. TCBV was transmitted from tulip to tulip and TV. clevelandii by the aphid Myzus persicae. Unlike TBV it was not transmitted to Lilium formosanum either by M. persicae or by manual inoculation. Leaf extracts from TCBV-containing TV. clevelandii were infective after dilution to l0^-3 but not 10-⁴ and after heating for 10 min at 50°C but not 60°C; infectivity and particle recovery were adversely affected by freezing at -20°C. TCBV particles were purified (c. 1 mg/100g g N. clevelandii leaf) from tissue extracts in 0·3 M citrate buffer containing 10 mM EDTA and 0·2% (v/v) 2-mercaptoethanol at pH 7·4 by clarification with 8·5% (v/v) n-butanol followed by differential centrifugation and sucrose density gradient centrifugation. Purified particles measured c. 720 × 12 nm. Virus particle antigen was readily detected in leaf and tepal extracts of tulip by enzyme-linked immunosorbent assay. A distant serological relationship was found between particles of TCBV and those of bean yellow mosaic virus but no serological relationship was found to TBV or four other potyviruses.     

Chemical Composition of Volatiles; Antimicrobial,Antioxidant and Cholinesterase Inhibitory Activity of Chaerophyllum aromaticum L. (Apiaceae) Essential Oils and Extracts

The present study reports the chemical composition of the headspace volatiles (HS) and essential oils obtained from fresh Chaerophyllum aromaticum root and aerial parts in full vegetative phase, as well as biological activities of their essential oils and MeOH extracts. In HS samples, the most dominant components were monoterpene hydrocarbons. On the other hand, the essential oils consisted mainly of sesquiterpenoids, representing 73.4% of the root and 63.4% of the aerial parts essential oil. The results of antibacterial assay showed that the aerial parts essential oil and MeOH extract have no antibacterial activity, while the root essential oil and extract showed some activity. Both of the tested essential oils exhibited anticholinesterase activity (47.65% and 50.88%, respectively); MeOH extract of the root showed only 8.40% inhibition, while aerial part extract acted as an activator of cholinesterase. Regarding the antioxidant activity, extracts were found to be more effective than the essential oils.     

Evaluation of antimicrobial activity of certain Chinese plants used in folkloric medicine

Six selected plants, belonging to 3 families from Nanjing of China, were extracted with the solvent 95% (v/v) ethanol to yield 11 extracts. The extracts were evaluated for their effects on the growth of eight clinical bacteria, two fungi and one yeast using a modified agar diffusion method. The results showed that the majority of the extracts investigated showed greater activities against the Gram-positive bacteria than against the Gram-negative bacteria, the fungi and the yeast. The strongest antimicrobial activity was exhibited by the stem extracts of Mahonia fortunei against multiresistant Staphylococcus aureus strains, followed by the stem extracts of Mahonia bealei, while Bacillus thuringiensis was the most sensitive to all extracts.     

Antimicrobial and Cytotoxic Activity of Extracts of Ferula heuffelii Griseb. ex Heuff. and Its Metabolites

The antimicrobial and cytotoxic activities of isolates (CHCl₃ and MeOH extracts and selected metabolites) obtained from the underground parts of the Balkan endemic plant Ferula heuffelii Griseb . ex Heuff . were assessed. The CHCl₃ and MeOH extracts exhibited moderate antimicrobial activity, being more pronounced against Gram‐positive than Gram‐negative bacteria, especially against Staphylococcus aureus (MIC=12.5 μg/ml for both extracts) and Micrococcus luteus (MIC=50 and 12.5 μg/ml, resp.). Among the tested metabolites, (6E)‐1‐(2,4‐dihydroxyphenyl)‐3,7,11‐trimethyl‐3‐vinyldodeca‐6,10‐dien‐1‐one ( 2 ) and (2S*,3R*)‐2‐[(3E)‐4,8‐dimethylnona‐3,7‐dien‐1‐yl]‐2,3‐dihydro‐7‐hydroxy‐2,3‐dimethylfuro[3,2‐c]coumarin ( 4 ) demonstrated the best antimicrobial activity. Compounds 2 and 4 both strongly inhibited the growth of M. luteus (MIC=11.2 and 5.2 μM , resp.) and Staphylococcus epidermidis (MIC=22.5 and 10.5 μM , resp.) and compound 2 additionally also the growth of Bacillus subtilis (MIC=11.2 μM ). The cytotoxic activity of the isolates was tested against three human cancer cell lines, viz., cervical adenocarcinoma (HeLa), chronic myelogenous leukemia (K562), and breast cancer (MCF‐7) cells. The CHCl₃ extract exhibited strong cytotoxic activity against all cell lines (IC₅₀<11.0 μg/ml). All compounds strongly inhibited the growth of the K562 and HeLa cell lines. Compound 4 exhibited also a strong activity against the MCF‐7 cell line, comparable to that of cisplatin (IC₅₀=22.32±1.32 vs. 18.67±0.75μM ).     

Cryopreservation of Chirostoma jordani sperm,fish model for the conservation of the genus in Mexico

Genus Chirostoma belongs to Atherinopsidae family and it is an endemic species from the Mesa Central in Mexico. Abundance of its species have decreased and some ones have been placed on the threatened species list, because of overfishing, urbanization, industrialization, destruction, habitat fragmentation, pollution and exotic species introduction. Chirostoma jordani (Woolman, 1894) is a freshwater fish with biological, ecological, cultural, and commercial importance. It has a broad distribution in Lerma drainage, Durango and Mexico City. In this last locality, their populations, although small, still persist in Xochimilco Lake; it is necessary to implement biotechnologies for their conservation, because of these causes and their basic biology. The aim was to standardize a sperm cryopreservation protocol in C. jordani, to determine extender solution, cryoprotective agent type and concentration, equilibrium time, freezing and thawed rate to be applied in assisted reproduction and conservation of genus Chirostoma. Chirostoma jordani adult males were collected in Atlangatepec Dam, Tlaxcala State, Mexico, to fresh seminal evaluation and cryopreservation protocol standardization. Four cryoprotectants effect was evaluated: dimethylsulfoxide (DMSO), methanol (MeOH), ethylene glycol (EG), and glycerol (GL) at five concentrations: 2%, 6%, 10%, 14% and 16% v/v. Higher and lower DMSO and MeOH 10% and EG 14%, decreased post-thaw motility percentage. Both DMSO and MeOH 10% and EG 14% had the highest post-thaw motility percentages, 48.8 ± 1.5%, 54.5 ± 1.0% and 53.5 ± 1.0%, at 15, 10, and 5 min equilibrium times, respectively, thawed at 40°C. Chirostoma jordani sperm can be cryopreserved with both DMSO and MeOH 10%, and EG 14%. These ones can be used for assisted reproduction. GL was not efficient, since it presented a post-thaw motility percentage very low.