Neutralisation of Local Haemorrhage Induced by the Saw-Scaled Viper Echis carinatus sochureki Venom Using Ethanolic Extract of Hibiscus aethiopicus L.

The objective of the study is to investigate the anti-snake venom activities of a local plant, Hibiscus aethiopicus L. The H. aethiopicus was dried and extracted with ethanol. Different assays were performed according to standard techniques, to evaluate the plant's acute toxicity and its antivenom activities. The results of evaluating the systemic acute toxicity of the H. aethiopicus extract using "oral and intra-peritoneal" route were normal even at the highest dose (24 g/kg) tested. All guinea pigs (n = 3) when treated with venoms E. c. sochureki (75 µg) alone induced acute skin haemorrhage. In contrast, all guinea pigs (n = 18) treated with both venom and the plant extract at a concentration between 500 and 1000 mg/kg showed no signs of haemorrhage. Moreover, all guinea pigs (n = 18) treated with venom and the plant extract below 400 mg/kg showed acute skin haemorrhage. All guinea pigs treated with venom E. c. sochureki (75 µg) alone induced acute skin haemorrhage after both 24 and 32 hours. In contrast, all guinea pigs treated with both venom and the plant extract (administered independently) at concentrations between 500 and 1000 mg/kg showed no signs of haemorrhage after 32 hours. However, after 24 hours all tested guinea pigs showed less inhibition (<60%) compared to that obtained after 32 hours. The outcome of this study reflects that the extract of H. aethiopicus plant may contain an endogenous inhibitor of venom induced local haemorrhage.

Antimicrobial and antioxidant activities of the various extracts of Verbascum pinetorum Boiss. O. Kuntze (Scrophulariaceae).

BACKGROUND AND OBJECTIVES: The present study was performed to evaluate the in vitro antimicrobial and antioxidant properties of various extracts of Verbascum (V.) pinetorum, a member of Scrophulariaceae family. While the antimicrobial activity of various extracts of V. pinetorum was determined with agar-well diffusion method, the antioxidant activity was examined with two complementary test systems, namely 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging and beta-carotene/linoleic acid test systems. RESULTS: The hexane extract exhibits antimicrobial activity against few microorganisms. However, dichloromethane, direct methanol and methanol/chloroform extracts are effective on a broad range of microorganisms. Among the tested bacteria Haemophilus influenzae was found to be the most sensitive bacterium. The 50% (IC50) inhibition activity of the methanolic extract of V. pinetorum on the free radical DPPH was determined as 13.04 mg/ml. In the case of the linoleic acid system, oxidation of linoleic acid was inhibited by methanolic extract of V. pinetorum, which showed 89.39% inhibition that is quite close to the value of the synthetic antioxidant reagent butylhydroxytoluene (BHT), 92.46%. Iridoid glycosides, flavonoids and saponins were determined as the major natural compounds in the methanolic extracts. The total phenolic components of V. pinetorum were found as 42.45 mg/g gallic acid equivalent. CONCLUSION: The results provide evidence that the extracts of V. pinetorum contained iridoid glycosides, flavonoids, saponins and phenolic compounds which may be responsible for the substantial antimicrobial and antioxidant activities.

Serine protease variants encoded by Echis ocellatus venom gland cDNA: cloning and sequencing analysis.

Envenoming by Echis saw-scaled viper is the leading cause of death and morbidity in Africa due to snake bite. Despite its medical importance, there have been few investigations into the toxin composition of the venom of this viper. Here, we report the cloning of cDNA sequences encoding four groups or isoforms of the haemostasis-disruptive Serine protease proteins (SPs) from the venom glands of Echis ocellatus. All these SP sequences encoded the cysteine residues scaffold that form the 6-disulphide bonds responsible for the characteristic tertiary structure of venom serine proteases. All the Echis ocellatus EoSP groups showed varying degrees of sequence similarity to published viper venom SPs. However, these groups also showed marked intercluster sequence conservation across them which were significantly different from that of previously published viper SPs. Because viper venom SPs exhibit a high degree of sequence similarity and yet exert profoundly different effects on the mammalian haemostatic system, no attempt was made to assign functionality to the new Echis ocellatus EoSPs on the basis of sequence alone. The extraordinary level of interspecific and intergeneric sequence conservation exhibited by the Echis ocellatus EoSPs and analogous serine proteases from other viper species leads us to speculate that antibodies to representative molecules should neutralise (that we will exploit, by epidermal DNA immunization) the biological function of this important group of venom toxins in vipers that are distributed throughout Africa, the Middle East, and the Indian subcontinent.

Antisnake Venom Activity of Hibiscus aethiopicus L. against Echis ocellatus and Naja n. nigricollis.

The objective of the study is to investigate whether the Hibiscus aethiopicus L. plant has neutralization activity against venoms of two clinically important snakes. The H. aethiopicus was dried and extracted with water. Different assays were performed to evaluate the plant's acute toxicity and its anti-snake venom activities. The results showed that H. aethiopicus extract alone had no effect on the viability of C(2)C(12) muscle cells, but significantly (P < .05) protected muscle cells against the toxic effects of E. ocellatus venom at 55, 150, and 300 mug/mL. The maximum protective effect of the extract was exhibited at 75 mug/mL. The extract significantly (P < .001) inhibited the cytotoxic effects of E. ocellatus venom at 300 mug/mL. All rabbits (n = 10) and guinea pigs (n = 10) were alive after the two weeks of given the lethal dosage 16 g/Kg of the H. aethiopicus extract herbal solution. No abnormal behaviour was observed of both groups of animals. All guinea pigs (n = 3) treated with venoms alone (5 mg/kg) died. However, all guinea pigs (n = 21) treated with venom (5 mg/kg) and the extract (400 to 1000 mg/kg) survived. Guinea pigs (n = 3) treated with Naja n. nigricollis venom alone (2.5 mg/kg) and guinea pigs (n = 21) venom with the extract (400 to 1000 mg/kg) died. The H. aethiopicus completely (100%) blocked the haemorrhagic activity of E. ocellatus in the egg embryo at 3.3 mg/mL of extract. These findings suggest that H. aethiopicus may contain an endogenous inhibitor of venom-induced haemorrhage.

Antimicrobial and cytotoxic abietane diterpenoids from the roots of Meriandera benghalensis (Roxb.) Benth.

Four abietane diterpenoids were isolated from the methanolic extract of the roots of Meriandera benghalensis and tested for their biological activity. Cryptotanshinone (2) and 17-hydroxycryptotanshinone (4) are known metabolites however the occurrence of tanshinone IIA (1) and przewaquinone A (3) from Meriandera benghalensis is reported for the first time. The four diterpenoids were identified by MS and one- and two dimensional NMR experiments. The isolated compounds were tested for their in vitro antiproliferative activity against three human cancer cell lines (a lung cancer (A-427), a urinary bladder cancer (5637) and a breast cancer (MCF-7) cell line) and for their antibacterial effect against three Gram-positive bacterial strains. All four abietanes showed potent cytotoxic effect against all cancer cell lines (IC50 between 1 and 8 microM) as well as antibacterial effect against the bacteria tested (MIC values between 33 and 70 microM).

Evaluation of the in vitro anticancer, antimicrobial and antioxidant activities of some Yemeni plants used in folk medicine.

The present research study deals with the evaluation of sixty four methanolic and aqueous extracts of thirty Yemeni plants used in traditional medicine for their in vitro antiproliferative activity against three human cancer cell lines in a microtiter plate assay based on cellular staining with crystal violet, for their antimicrobial activity against antibiotic susceptible three Gram-positive, three Gram-negative bacterial and one fungal stains and three multiresistant Staphylococcus strains by the agar diffusion method and the determination of MIC against three Gram-positive bacteria with the broth micro-dilution assay, as well as for their antioxidant activity using the DPPH radical scavenging method. Furthermore the chemical composition of the methanolic extracts was determined by using chromatographic methods. As a result of this work, 12 Yemeni herbs namely Centaurothamus maximus, Costus arabicus, Cupressus sempervirens, Dichrocephala integrifolia, Euphorbia schimperi, Gomphocarpus fruticosus, Kanahia laniflora, Meriandera benghalensis, Pulicaria inuloides, Solanum glabratum, Tarconanthus camphoratus and Vernonia leopoldii demonstrated a noteworthy growth inhibitory effect against all cancer cell lines with IC50 values <50 microg/ml. Pronounced antimicrobial activity was observed only against Gram-positive bacteria among them multiresistant bacteria with inhibition zones >15 mm and MIC values <500 microg/ml, by 9 plants especially Centaurothamus maximus, Cupressus sempervirens, Enicostemma verticillare, Meriandera benghalensis, Nepeta deflersiana, Pulicaria inuloides, Tarconanthus camphoratus, Teucrium yemense and Vernonia leopoldii. Moreover, the methanolic extracts of Cupressus sempervirens, Meriandera benghalensis, Pulicaria inuloides and Rhus retinorrhaea showed a remarkable radical scavenging effect at low concentrations.

Study of the anticancer potential of Yemeni plants used in folk medicine.

The present work evaluated the anticancer activity of methanol extracts from 24 plants used in Yemeni traditional medicine. To evaluate the in vitro cytotoxic potency of the investigated extracts, an established microtiter plate assay based on cellular staining with crystal violet was used with 5 human cancer cell lines: two lung cancer (A-427 and LCLC-103H), two urinary bladder carcinoma (5637 and RT-112) and one breast cancer (MCF-7) line. The methanolic extracts of Dendrosicyos socotrana, Withanina aduensis, Withania riebeckii, Dracena cinnabari and Buxus hildebrandtii exhibited the highest toxicity on all tumor cell lines with IC50 values ranging between 0.29 and 5.54 microg/ml. The extracts of Jatropha unicostata and Punica protopunica showed a moderate potency on the most tumor cell lines.

Screening of selected Arabian medicinal plant extracts for inhibitory activity against peptidases.

The methanolic extracts of 20 medicinal plants from the island Soqotra/Yemen were screened with respect to their inhibitory potency against angiotensin converting enzyme (ACE), neutral endopeptidase (NEP) and aminopeptidase N (APN). Eight extracts did not show significant inhibitory activity against the enzymes tested, only Kalanchoe farinacea, Boswellia elongata and Cissus hamaderohensis inhibited all three enzymes. The most active extract was prepared from Kalanchoe farinacea characterized by low IC50 values especially for NEP and APN.

Antiviral activity of Inonotus hispidus.

The antiviral activity of two phenolic compounds, hispolon and hispidin, isolated from the fruit bodies of the basidiomycete Inonotus hispidus as well as of some extracts prepared from fruit bodies and mycelial cultures of this fungus was investigated in allantois on the shell-test system. Ethanolic extracts, hispidin and hispolon showed considerable antiviral activity against influenza viruses type A and B.

Antiviral lanostanoid triterpenes from the fungus Ganoderma pfeifferi.

Ganodermadiol, lucidadiol and applanoxidic acid G were isolated as first triterpenes from the European Basidiomycete Ganoderma pfeifferi. The compounds show antiviral activity against influenza virus type A and HSV type 1.

Ganomycins A and B, new antimicrobial farnesyl hydroquinones from the basidiomycete Ganoderma pfeifferi.

Two new farnesyl hydroquinones named ganomycin A (1) and ganomycin B (2) were isolated from Ganoderma pfeifferi, and their structures were elucidated by spectroscopic methods. Both carboxylic acids exhibit antimicrobial activity against several Gram-positive and Gram-negative bacteria.