Hasan Soliman Yusufoglu

Effect of Emex spinosa, Leptadenia pyrotechnica, Haloxylon salicornicum and Ochradenus baccatus extracts on the reproductive organs of adult male rats.

Context: Emex spinosa (L.) Campd. (Polygonaceae), Leptadenia pyrotechnica (Forsk.) Decne (Asclepiadaceae), Haloxylon salicornicum (Moq.) Bunge ex Bioss. (Chenopodiaceae) and Ochradenus baccatus Delile (Resedaceae) are used in folk medicine for treatment of male sexual disorders. Objective: To investigate the effects of E. spinosa, L. pyrotechnica, H. salicornicum and O. baccatus extracts on the reproductive system of male rats after prolonged period of treatment. Methods: Seventy-eight healthy adult male Wistar rats were divided into 13 groups (6 animals, each). The plant extracts (100, 200 and 400 mg/kg) were given daily by gavage to different groups of rats for 65 days. The thirteenth group (control) received the vehicle only. Test and control rats were mated with estrus female rats on days 30, 45 and 60 of treatment. Body and relative reproductive organ weights, and sperm parameters were recorded. Results: Animals treated with the ethanol extracts of E. spinosa and L. pyrotechnica showed significant improvement of the relative weight of reproductive organs, sperm count, sperm motility and total sperm abnormality. The mean sperm count for E. spinosa group (400 mg/kg) was 233.7 ± 4.50 × 106/mL, for L. pyrotechnica (200 and 400 mg/kg) groups were 237.0 ± 5.22 × 106/mL and 240.3 ± 4.64 × 106/mL, respectively and that of the control group was 218.1 ± 4.28 × 106/mL. The sperm motility of the control group was 77.5 ± 2.12, those of E. spinosa (400 mg/kg) group was 87.3 ± 3.50% and those of L. pyrotechnica (200 and 400 mg/kg) groups were 86.0 ± 3.11 and 89.7 ± 2.90%, respectively. Ethanol extracts of E. spinosa (400 mg/kg) and L. pyrotechnica (200 and 400 mg/kg) significantly elevate the serum levels of testosterone (5.30 ± 0.15, 5.32 ± 0.20 and 5.66 ± 0.19 ng/mL, respectively vs 4.64 ± 0.16 ng/mL) and luteinizing hormone (0.69 ± 0.03, 0.70 ± 0.03 and 0.74 ± 0.03 mIU/mL, respectively vs 0.59 ± 0.02 mIU/mL). On the other hand, no alterations were observed in body and relative organ weights, sperm numbers as well as sperm morphology of the male rats after the exposure to the H. salicornicum and O. baccatus extracts for 65 days. Conclusions: E. spinosa and L. pyrotechnica extracts appear to possess fertility improvement activity in male rats due to their testosterone increasing property. Moreover, the results suggest the absence of male reproductive toxicity of the H. salicornicum and O. baccatus extracts at tested doses.

Chemical Composition and Biological Activity of Haplophyllum tuberculatum Juss. Essential Oil

Abstract: The essential oil of Haplophyllum tuberculatumwas prepared by hydrodistillation of the fresh flowering aerial parts of the plant collected from Saudi Arabia. The oil was subsequently analyzed by GC and GC-MS. Thirty seven compounds, accounting for 96.4 % of the oil composition were identified. The major components were trans-p-menth-2-en-1-ol (19.2 %), cis-p-menth-2-en-1-ol (13.2 %), myrcene (10.1 %), δ-3- carene (8.8 %), β-phellandrene (6.9 %), limonene (6.6 %) and cis-piperitol (6.4 %). The antimicrobial activity of the essential oil was determined using the broth microdilution method against various human pathogens, where a relatively low inhibitory range was observed (MIC 1 mg/mL). Furthermore, the oil was evaluated for its antifungal activity against the strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides using the direct overlay bioautography assay. The essential oil showed no antifungal activity at 80 and 160 µg/spot concentrations compared to commercial antifungal standards. The oil was also investigated for its insecticidal and repellent activity against Aedes aegypti. The oil was repellent to the yellow fever mosquito Ae. aegypti using the “cloth patch assay” down to a concentration of 0.074 mg/cm2; however, the oil had low toxicity against first instar larvae and adults of Ae. aegypti in a high throughput larval bioassay and adult topical assay. Additionally, enzyme activity was measured using the spectrophotometric Ellman method. The oil showed weakly acetylcholinesterase (AChE) inhibitory activity at the tested concentration, compared to standard substances, whereas no inhibition on butyrylcholinesterase (BuChE) activity was observed.

Analgesic, antipyretic, anti-inflammatory, hepatoprotective and nephritic effects of the aerial parts of Pulicaria arabica (Family: Compositae) on rats

OBJECTIVE To explore the analgesic, antipyretic, anti-inflammatory, hepatic and nephritic effects of Pulicaria arabica (P. arabica) in several experimental models. METHODS For analgesic effect hot plate and writhing method were used, while for antipyretic and anti-inflammatory rectal temperature and carrageenan induced hind paw edema were used respectively. CCl4 intoxication method was used for hepatic and nephritic protective activity. RESULTS The results of the present studies revealed that P. arabica has potent analgesic, antipyretic and anti-inflammatory with the significant hepatic and nephritic protecting actions. The CCl4 intoxication changed the normal malondialdehyde and nonprotein sulfhydryls levels in both liver and kidney. These changes were normalized with P. arabica indicating the antioxidant nature of this plant. CONCLUSIONS The results of present study indicated that P. arabica can be used in analgesic, antipyretic and anti-inflammatory conditions even in hepatic and nephritic conditions. More supportive studies are required before clinical recommendation.

HPTLC densitometric method for analysis of thymoquinone in Nigella sativa extracts and marketed formulations

Objective To develop a sensitive and accurate high-performance thin layer chromatography method and to determine the quantity of thymoquinone in in two different N. sativa extracts and marketed formulations.

Rhanterium epapposum Oliv. essential oil: Chemical composition and antimicrobial, insect-repellent and anticholinesterase activities

The essential oil (EO) of the aerial parts of Rhanterium epapposum Oliv. (Asteraceae), was obtained by hydrodistillation. The oil was subsequently analyzed by both GC-FID and GC-MS, simultaneously. Forty-five components representing 99.2% of the oil composition were identified. The most abundant compounds were camphene (38.5%), myrcene (17.5%), limonene (10.1%) and α-pinene (8.7%). Referring to the ethnobotanical utilization, an insecticidal assay was performed, where the oil repelled the yellow fever mosquito Aedes aegypti L. at a minimum effective dose (MED of 0.035 ± 0.010 mg/cm2) compared to the positive control DEET (MED of 0.015 ± 0.004 mg/cm2). Additionally, the in vitro antimicrobial activity against a panel of pathogens was determined using a microdilution method. The acetyl- and butyrylcholine esterase inhibitory activities were measured using the colorimetric Ellman method. The bioassay results showed that the oil was rather moderate in antimicrobial and cholinesterase inhibitions when compared to the standard compounds.

Secondary metabolites from Astragalus karjaginii BORISS and the evaluation of their effects on cytokine release and hemolysis

A new cycloartane sapogenol and a new cycloartane xyloside were isolated from Astragalus karjaginii BORISS along with thirteen known compounds. The structures of the new compounds were established as 3-oxo-6α,16β,24(S),25-tetrahydroxycycloartane (1) and 6-O-β-d-xylopyranosyl-3β,6α,16β,24(S),25-pentahydroxycycloartane (2) by 1D- and 2D-NMR experiments as well as ESIMS and HRMS analyses. The presence of the keto function at position 3 was reported for the first time for cyclocanthogenol sapogenin of Astragalus genus. In vitro immunomodulatory effects of the new compounds (1 and 2) along with the n-BuOH and MeOH extracts of A. karjaginii at two different doses (3 and 6μg) were tested on human whole blood for in vitro cytokine release (IL-2, IL-17A and IFN-γ) and hemolytic activities. The results confirmed that compound 2, a monodesmosidic saponin, had the strongest effect on the induction of both IL-2 (6μg, 6345.41±0.12pg/mL (×5), P<0.001) and a slight effect upon IL-17A (3μg, 5217.85±0.72pg/mL, P<0.05) cytokines compared to the other test compounds and positive controls (AST VII: Astragaloside VII; and QS-21: Quillaja saponin 21). All tested extracts and molecules also induced release of IFN-γ remarkably ranging between 5031.95±0.05pg/mL, P<0.001 for MeOH extract (6μg) and 5877.08±0.06pg/mL, P<0.001 for compound 1 (6μg) compared to QS-21 (6μg, 5924.87±0.1pg/mL, P<0.001). Administration of AST VII and other test compounds did not cause any hemolytic activity, whereas QS-21 resulted a noteworthy hemolysis.

New cytotoxic sesquiterpene lactones from Anthemis scrobicularis

Four new sesquiterpene lactones, 4α-hydroxy-guaia-10(14),11(13)-diene-12,6α-olide (1), 4α-hydroxy-9α-acetoxy-guaia-1(10),2-diene-12,6α-olide (4), 4α-hydroxy-9β-acetoxy-guaia-1(10),2-diene-12,6α-olide (5), and 1α,4α-dihydroxy-9α-acetoxy-guaia-10(14),2-diene-12,6α-olide (6), were isolated from the aerial parts of Anthemis scrobicularis. Their structures were elucidated on the basis of their IR, NMR, and MS spectroscopic data. In addition, two known sesquiterpene lactones micheliolide (2) and achillin (3) were also isolated. The cytotoxicity of some of the isolated compounds was tested against HCT 116, HepG-2, and MCF-7 cell lines. Micheliolide and 4α-hydroxy-guaia-10(14),11(13)-diene-12,6α-olide showed pronounced inhibitory activity while 4α-hydroxy-9α-acetoxy-guaia-1(10),2-diene-12,6α-olide showed weak activity.

Mosquito and tick repellency of two Anthemis essential oils from Saudi Arabia

The essential oils (EOs) of Anthemis melampodina (Am) and Anthemis scrobicularis (As) (Asteraceae) were extracted from the aerial parts of the plants by hydrodistillation, and their chemical compositions were analyzed using GC-FID and GC-MS. Fifty-six components representing 85.5% of the oil composition of Anthemis melampdina were identified, and the major components were α-pinene (17.1%) and β-eudesmol (13.8%). Forty-one components representing 86% of the oil composition of Anthemis scrobicularis were identified, and the major component was β-eudesmol (12.8%). Laboratory bioassays were conducted to determine repellency of Am and As EOs against the yellow fever mosquito Aedes aegypti L. and the lone star tick Amblyomma americanum L. The minimum effective doses (MEDs) of the Am and As EOs against mosquitoes were 0.187 ± 0.000 and 0.312 ± 0.063 mg/cm2 respectively, which were significantly higher than that of DEET (0.023 ± 0.000 mg/cm2) in human-based repellent bioassays. The As EO was more repellent than Am EO against nymphal ticks but was less effective than DEET in vertical paper bioassays.

Investigation of antioxidant compounds in commercial pomegranate molasses products using matrix-solid phase dispersion extraction coupled with HPLC

Pomegranate is a well known fruit for its unique flavor, taste and health benefits. The medicinal properties of this fruits directly associated with the phenolic content present, with great anti-oxidant potential. The research is intended to develop matrix solid phase dispersion method (MSPD) and HPLC quantification of four major anti-oxidant marker constituents (vitamin C, gallic acid, rutin & ellagic acid) in pomegranate molasses samples. The effects of several important experimental parameters like type of dispersant, sample-dispersant ratio, solvents and its volume, time of extraction were investigated. A C18 column with the specification (5 µm, 250 × 4.0 mm) was used for the separation. A gradient flow of mobile phase was selected after many trials containing 0.1%, v/v solution of orthophosphoric acid and acetonitrile. The flow rate was 1.0 mL/min; and the chromatograms were recorded at 254 nm. The validation parameters, like linearity (r2 = 0.9985, 0.9965, 0.9925 & 0.9986), accuracy (100.3, 99.5, 100.9 & 101.9%), intra-day precision (%RSD = 1.09, 1.02, 1.26 & 0.97), inter-day precision (%RSD = 1.32, 0.83, 1.07, & 1.15) LOD (0.07, 4.50, 0.45 & 0.40 µg/mL), LOQ (0.095, 9.50, 0.85 & 9.5 µg/mL) and robustness (% RSD = 0.92, 0.76, 0.81 & 0.83) respectively for vitamin C, gallic acid, rutin & ellagic acid, were found satisfactory as per ICH guidelines.

Biotransformation of Neoruscogenin by the Endophytic Fungus Alternaria eureka

Biotransformation of neoruscogenin (NR, 1, spirosta-5,25(27)-diene-1β,3β-diol), the major bioactive sapogenin of Ruscus preparations, was carried out with the endophytic fungus Alternaria eureka. Fourteen new biotransformation products (2-15) were isolated, and their structures were elucidated by NMR and HRESIMS data analyses. A. eureka affected mainly oxygenation, oxidation, and epoxidation reactions on the B and C rings of the sapogenin to afford compounds 8-15. In addition to these, cleavage of the spiroketal system as in compounds 2-7 and subsequent transformations provided unusual metabolites. This is the first study reporting conversion of the spirostanol skeleton to cholestane-type metabolites 2-5. Additionally, the cleavage of the C-22/C-26 oxygen bridge yielding a furostanol-type steroidal framework and subsequent formation of the epoxy bridge between C-18 and C-22 in 7 was encountered for the first time in steroid chemistry.