In vitro anti-malarial interaction and gametocytocidal activity of cryptolepine.

BACKGROUND: Discovery of novel gametocytocidal molecules is a major pharmacological strategy in the elimination and eradication of malaria. The high patronage of the aqueous root extract of the popular West African anti-malarial plant Cryptolepis sanguinolenta (Periplocaceae) in traditional and hospital settings in Ghana has directed this study investigating the gametocytocidal activity of the plant and its major alkaloid, cryptolepine. This study also investigates the anti-malarial interaction of cryptolepine with standard anti-malarials, as the search for new anti-malarial combinations continues. METHODS: The resazurin-based assay was employed in evaluating the gametocytocidal properties of C. sanguinolenta and cryptolepine against the late stage (IV/V) gametocytes of Plasmodium falciparum (NF54). A fixed ratio method based on the SYBR Green I fluorescence-based assay was used to build isobolograms from a combination of cryptolepine with four standard anti-malarial drugs in vitro using the chloroquine sensitive strain 3D7. RESULTS: Cryptolepis sanguinolenta (IC50 = 49.65 nM) and its major alkaloid, cryptolepine (IC50 = 1965 nM), showed high inhibitory activity against the late stage gametocytes of P. falciparum (NF54). In the interaction assays in asexual stage, cryptolepine showed an additive effect with both lumefantrine and chloroquine with mean ΣFIC50s of 1.017 ± 0.06 and 1.465 ± 0.17, respectively. Cryptolepine combination with amodiaquine at therapeutically relevant concentration ratios showed a synergistic effect (mean ΣFIC50 = 0.287 ± 0.10) whereas an antagonistic activity (mean ΣFIC50 = 4.182 ± 0.99) was seen with mefloquine. CONCLUSIONS: The findings of this study shed light on the high gametocytocidal properties of C. sanguinolenta and cryptolepine attributing their potent anti-malarial activity mainly to their effect on both the sexual and asexual stages of the parasite. Amodiaquine is a potential drug partner for cryptolepine in the development of novel fixed dose combinations.

Mosquito larvicidal activity of Cassia tora seed extract and its key anthraquinones aurantio-obtusin and obtusin.

BACKGROUND: The edible and medicinal leguminous plant Cassia tora L. (Fabaceae) is known to possess insecticidal properties against a wide range of plant-feeding insects. However, the bioactivity of extracts of this plant and their constituents against vectors of medical importance has been largely unexplored. We investigated the mosquito larvicidal activity of the seed extract and its major anthraquinones against larvae of the African malaria vector Anopheles gambiae (s.s.). METHODS: Third-fourth instar larval mortality was observed after 24, 48, 72 and 96 h of exposure to varying doses of the extracts, and two anthraquinones isolates identified using liquid chromatography- quadrupole time of flight mass spectrometry (LC-QtoF-MS). The mosquito larval mortality was evaluated relative to the natural insecticide azadirachtin. RESULTS: Fractionation of the crude extract decreased mosquito larvicidal activity, however, larvicidal activity increased with increasing dose of the treatment and exposure time. The known anthraquinones aurantio-obtusin and obtusin were identified as key larvicidal compounds. Aurantio-obtusin and obtusin, exhibited similar toxicity to larvae of A. gambiae (s.s.) with LD50 values of 10 and 10.2 ppm, respectively. However, the two anthraquinones were four- and ~ six-fold less potent than that of the crude seed extract and azadirachtin, which had comparable LD50 values of 2.5 and 1.7 ppm, respectively. CONCLUSION: Both aurantio-obtusin and obtusin showed mosquito larvicidal activity which were comparable to their respective fractions although they were less potent relative to the crude extract and azadirachtin. Further studies need to be conducted on C. tora for its exploitation as a potential eco-friendly tool in mosquito larval source reduction.

Antinflammatory and Analgesic Effects in Rodent Models of Ethanol Extract of Clausena anisata Roots and their Chemical Constituents.

The in vivo antiinflammatory and analgesic activities of the crude ethanol extract and chemical constituents of Clausena anisata roots were investigated. The crude extract, which was devoid of any visible acute toxicity, displayed significant antiinflammatory effect at the dose of 1000 mg/kg (p.o.) when assessed using the carrageenan-induced oedema model. In the acetic acid-induced writhing and hot plate tests, it produced a very significant (p < 0.001); dose- dependent analgesic effect, with maximum analgesic activity of 72.1% at 1000 mg/kg (p.o.). Phytochemical analysis of the crude extract resulted in the isolation of four coumarins (anisocoumarin B, osthol, imperatorin and xanthotoxol) and a carbazole alkaloid, heptaphylline. Among the isolated compounds, osthol and anisocournarin B produced the highest antiinflammatory activity at 9 mg/kg (p.o.): slightly better than the positive control, indomethacin. Except for xanthotoxol, all the isolated compounds administered at 6 mg/kg (p.o.) produced significant analgesic activity and higher than diclofenac; with- heptaphylline being the most potent (48.7%). The analgesic activity of anisocoumarin B (50.4%) was the highest among the isolates tested and the standard, tramadol, in the hot plate test. The nonselective opioid receptor antagonist, naloxone, abolished the analgesic effect of the crude extract and the tested isolates (anisocoumarin B and xanthotoxol) in the hot plate test suggesting an effect via the central opioidergic system. These findings provide the scientific basis for the use of C. anisata roots in traditional medicine as antiinflammatory and analgesic agents.

Medicinal plants and finished marketed herbal products used in the treatment of malaria in the Ashanti region, Ghana.

ETHNOPHARMACOLOGICAL RELEVANCE: Ethnobotanical survey was performed to document medicinal plants employed in the management of malaria in the Bosomtwe and Sekyere East Districts of the Ashanti Region (Ghana), in comparison with the plant ingredients in herbal antimalarial remedies registered by the Ghana Food and Drug Administration. MATERIALS AND METHODS: Two hundred and three (203) herbalists from 33 communities within the two districts were interviewed on medicinal plants they use to manage malaria. A literature search was made to determine already documented plants. In addition, 23 finished marketed herbal products indicated for the management of malaria were identified and their labels examined to find out which of the plants mentioned in our survey were listed as ingredients and whether these products are in anyway regulated. RESULTS: Ninety-eight (98) species of plants were cited for the management of malaria. In comparison with literature citations, 12 (12.2%) species were reported for the management of malaria for the first time and 20 (20.4%) others for the first time in Ghana. Twenty-three (23) finished marketed herbal antimalarial products examined contained aerial or underground parts of 29 of the plants cited in our survey as ingredients. Twenty-two (22) of these products have been registered by the Ghana Food and Drugs Authority, four (4) of which were included in the recommended herbal medicine list for treating malaria in Ghana. CONCLUSION: This study provides new additions to the inventory of medicinal plants used for the management of malaria and reports the commercial availability and regulation of finished marketed labelled herbal products intended for the treatment of malaria in Ghana.

Erythroivorensin: A novel anti-inflammatory diterpene from the root-bark of Erythrophleum ivorense (A Chev.).

The stem- and root-bark of Erythrophleum ivorense (A Chev., family, Fabaceae) are routinely employed in the West African traditional medicine to treat inflammation and a variety of other disease conditions. Although the chemistry and pharmacology of cassaine-type diterpene alkaloids isolated from the stem-bark of the plant are fairly established, the root-bark has not yet been investigated. In the present study, the crude aqueous-alcohol extract of the root-bark was demonstrated to display a time- and dose (30-300 mg/kg p.o.)-dependent anti-inflammatory effect in chicks. Comprehensive chromatographic analysis coupled with spectroscopic and X-ray study further allowed the assignment of one of the major anti-inflammatory constituents as a novel cassaine-type diterpene, erythroivorensin. The other major constituents were known anti-inflammatory compounds: a triterpene, betulinic acid and a flavonoid, eriodictyol. The dose (10-100mg/kg p.o.)-dependent anti-inflammatory effects of the three compounds were either comparable or more significant than the positive control, diclofenac.

A novel triterpene from the roots of Paullinia pinnata: 6α-(3'-methoxy-4'-hydroxybenzoyl)-lup-20(29)-ene-3-one.

Paullinia pinnata L. (Sapindaceae) is an endemic West African plant that is extensively used in traditional medicine to treat various diseases. Previous phytochemical analysis by various groups led to the isolation of several novel lupene-based triterpenene derivatives along with other classes of compounds. As part of our continued phytochemical studies on the roots of this plant, we have now identified yet another novel triterpene, 6a-(3'-methoxy-4'-hydroxybenzoyl)-lup-20(29)-ene-3-one. The identification of the compound through comprehensive spectroscopic studies is discussed.

Clerodane diterpenes from Polyalthia longifolia (Sonn) Thw. var. pendula: Potential antimalarial agents for drug resistant Plasmodium falciparum infection.

BACKGROUND: Plasmodium falciparum drug resistance is a major public health challenge in sub-Sahara Africa. Many people are now resorting to the use of herbs in managing malaria due to the increasing treatment failures with the conventional drugs. In this study the ethanolic extract of Polyalthia longifolia (Sonn) Thw. var. pendula, a variety fondly used in folklore medicine in Ghana was investigated for potential antimalarial drug development. METHOD: The ethanolic extract of P. longifolia (Sonn) Thw. var. pendula stem bark was screened against the multidrug resistant, K1 strain of P. falciparum by the parasite lactate dehydrogenase (pLDH) assay and a good antiplasmodial activity (IC50 22.04± 4.23µg/ml) was observed which led to further chromatographic analysis in search for actives. RESULTS: Bioassay guided fractionation of the extract yielded; three clerodane diterpenes [16-hydroxycleroda-3,13-dien-16,15-olide (1), 16-oxocleroda-3,13E-dien-15-oic acid (2) and 3,16-dihydroxycleroda-4(18),13(14)Z-dien-15,16-olide (3)], a steroid [beta-stigmasterol (4)] and two alkaloids [darienine (5) and stepholidine (6)]. While compounds 4, 5 and 6 exhibited weak antiplasmodial activity (IC50 22-105µg/ml), the clerodane diterpenes exhibited significantly potent (p<0.005) blood schizonticidal activity (IC50: 3-6µg/ml). This is the first report of the antiplasmodial activity of compounds 2 and 3. In combination assay with chloroquine, compounds 1, 2, 3 and 5 antagonized the antiplasmodial activity of chloroquine while 4 and 6 demonstrated a synergistic action. CONCLUSION: The potent antiplasmodial activity of the extract of P. longifolia (Sonn) Thw. var. pendula and compounds therein strongly suggests its usefulness as an antimalarial agent and supports its inclusion or exploitation in formulations of herbal remedies for malaria in Ghana.

Flavonoid glycosides from the stem bark of Margaritaria discoidea demonstrate antibacterial and free radical scavenging activities.

One new flavonoid glycoside, along with three known flavonoid glycosides were isolated from the stem bark of Margaritaria discoidea, which is traditionally used in the management of wounds and skin infections in Ghana. The new flavonoid glycoside was elucidated as hydroxygenkwanin-8-C-[α-rhamnopyranosyl-(1 → 6)]-ß-glucopyranoside (1) on the basis of spectroscopic analysis. The isolated compounds demonstrated free-radical scavenging as well as some level of antibacterial activities. Microorganisms including Staphylococcus aureus are implicated in inhibiting or delaying wound healing. Therefore, any agent capable of reducing or eliminating the microbial load present in a wound as well as decreasing the levels of reactive oxygen species may facilitate the healing process. These findings therefore provide some support to the ethnopharmacological usage of the plant in the management of wounds.

The heavy metal contents of some selected medicinal plants sampled from different geographical locations.

BACKGROUND: The levels of 5 minerals namely; lead, arsenic, mercury, cadmium, and aluminum were assessed in 10 medicinal plants sampled from 5 different geographical locations to determine the effect of location on the plants' mineral content. MATERIALS AND METHODS: Atomic absorption spectrophotometry (wet digestion) was used for the analyzes, and content of the minerals per sample was expressed as µg/g. The levels of minerals were compared to their limit specification for herbs and daily total intake of these minerals. A two-way analysis of variance, which tends to look at the effect of the location and the medicinal plant itself on the plants mineral content, was used in the statistical analysis. RESULTS: Lead (Pb) was present in all plant species examined, except Ocimum gratissimum. One plant exceeded the maximum safety limit for lead. Cadmium was also detected in some of the medicinal plant species (44%) whilst majority were below the detection limit (0.002) representing 56%. 40% of the plant species exceeded the limit for cadmium. Mercury and arsenic in all the plant species were below the detection limit (0.001). Significant variation existed in mineral content for the various locations (P ≤ 0.05). CONCLUSION: The findings generally suggest the variation in mineral levels for the various locations. Thus, our study has shown that same species of medicinal plants, growing in different environments, accumulates different levels of heavy metals.

In vitro anti-plasmodial activity of three herbal remedies for malaria in Ghana: Adenia cissampeloides (Planch.) Harms., Termina liaivorensis A. Chev, and Elaeis guineensis Jacq.

BACKGROUND: Herbal remedies of Adenia cissampeloides, Terminalia ivorensis, and Elaeis guineensis among others have been used in Ghana for the treatment of various ailments including malaria. However, most of these remedies have not been scientifically investigated. OBJECTIVE: This study, therefore, seeks to investigate the anti-plasmodial activity of these plants. MATERIALS AND METHODS: The ethanolic extracts of A. cissampeloides stem, T. ivorensis stem bark, and E. guineensis leaves were tested for in vitro anti-plasmodial activity against chloroquine-resistant strains of Plasmodium falciparum. Thin blood films were used to assess the level of parasitemia and growth inhibition of the extracts. RESULTS: The IC (50) of A. cissampeloides, T. ivorensis, and E. guineensis were 8.521, 6.949, and 1.195 µg/ml, respectively, compared to artesunate with IC(50) of 0.031 µg/ml. CONCLUSION: The result of this study appears to confirm the folkloric anti-malarial use these plants.

Antibiotic producing microorganisms from River Wiwi, Lake Bosomtwe and the Gulf of Guinea at Doakor Sea Beach, Ghana.

BACKGROUND: Microorganisms have provided a wealth of metabolites with interesting activities such as antimicrobial, antiviral and anticancer. In this study, a total of 119 aquatic microbial isolates from 30 samples (taken from water bodies in Ghana) were screened by the agar-well diffusion method for ability to produce antibacterial-metabolites. RESULTS: Antibacterial activity was exhibited by 27 of the isolates (14 bacteria, 9 actinomycetes and 4 fungi) against at least one of the indicator microorganisms: Enterococcus faecalis (ATCC 29212), Bacillus thuringiensis (ATCC 13838), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Proteus vulgaris (NCTC 4635) and Bacillus Subtilis (NCTC 10073). A sea isolate MAI2 (identified as a strain of Pseudomonas aeruginosa) exhibited the highest antibacterial activity (lowest zone of inhibition = 22 mm). The metabolites of MAI2 extracted with chloroform were stable to heat and gave minimum inhibitory concentrations ranging between 250 and 2000 µg/ml. Bioautography of the extract revealed seven active components. CONCLUSION: This study has therefore uncovered the potential of water bodies in the West African sub-region as reservoirs of potent bioactive metabolite producing microorganisms.

Traditional herbalists and cancer management in Kumasi, Ghana.

Cancer incidence rates are increasing in sub-Saharan Africa where traditional medical practitioners (TMPs) are involved in cancer management. Little is known about the specific role that TMPs play in cancer management in Ghana; we hypothesize that an understanding of the practices of TMPs with regard to cancer patients would help to enhance literacy about cancer amongst TMPs and would contribute to the diagnosis of cancer at earlier stages, by avoiding the detrimental delays while enlisting their help in certain activities that enhance cancer care. To elucidate the nature of the involvement of TMPs in cancer management, we conducted semi-structured interviews with 42 TMPs who practice in Kumasi, Ghana. The interviews elicited information about their knowledge and practices regarding cancer management and interactions with local hospitals. The results showed that TMPs tended to identify cancers as diseases of visible masses, fungating lesions, ulceration, and bleeding reflecting the advanced stages and types of cancers they usually encounter. TMPs identified certain causes of cancer and believed that they can treat and prevent cancer. These results indicate that TMPs are significant health service delivery resources in Ghana for patients potentially affected with cancer. Our work suggests that dedicated efforts to further integrate TMPs into the overall health care system would be beneficial to patients. Future research should examine the role of cancer education and training programs for TMPs to enhance their knowledge, strengthen their ability to complement allopathic practitioners, and increase early detection and treatment efforts through appropriate and timely referrals.

Acaricidal effect of an isolate from Hoslundia opposita vahl against Amblyomma variegatum (Acari: Ixodidae).

BACKGROUND: Hoslundia opposita Vahl. (Lamiaceae), a common local shrub in Ghana, is traditionally known not only for its pharmacological benefits but also for its insecticidal properties. Its acaricidal property, however, has not been investigated. OBJECTIVE: To test the acaricidal effects of the crude extract and fractions of H. opposita leaves as well as to isolate and characterize the acaricidal principles. MATERIALS AND METHODS: The crude methanolic extract, pet. ether, ethyl acetate and aqueous fractions of the leaves of H. opposita were tested against the larvae of the cattle tick, Amblyomma variegatum, using the Larval Packet Test. A bioassay-guided isolation was carried out to identify the acaricidal principle obtained from the ethyl acetate fraction. RESULTS: The active principle was characterised as ursolic acid, a triterpene previously isolated from the leaves of the same plant. The extract and fractions were less potent than the control, malathion (LC(50) 1.14 × 10(-4) mg/ml). Among the plant samples however the crude methanolic extract exhibited the highest effect against the larvae (LC(50) 5.74 × 10(-2) mg/ml), followed by the ethyl acetate fraction (LC(50) 8.10 × 10(-2) mg/ml). Ursolic acid, pet. ether and aqueous fractions however showed weak acaricidal effects with LC(50) values of 1.13 mg/ml, 8.96 × 10(-1) mg/ml and 1.44 mg/ml, respectively. CONCLUSION: Ursolic acid was not as potent as the crude methanolic extract and the ethyl acetate fraction from which it was isolated. The overall acaricidal effect of H. opposita may have been due to synergy with other principles having acaricidal properties.

Two novel lupane triterpenoids from Paullinia pinnata L. with fibroblast stimulatory activity.

OBJECTIVES: Novel lupane triterpenoids from Paullinia pinnata L., a Ghanaian plant traditionally used for wound healing, were examined for in-vitro fibroblast stimulatory activity using the 142BR cell line. METHODS: Bioactivity-guided isolation of the crude extract of P. pinnata L. was carried out in order to determine the nature of the compounds responsible for the stimulation of fibroblast proliferation. KEY FINDINGS: Two novel compounds were isolated and characterised, namely, 6beta-(3'-methoxy-4'-hydroxybenzoyl)-lup-20(29)-ene-one (1) and 6beta-(3'-methoxy-4'-hydroxybenzoyl)-lup-20(29)-ene-ol (2), together with three known compounds, friedelin (3), beta-sitosterol (4) and beta-sitosterol-3-D-glucoside (5). The methanol extract of the roots of P. pinnata caused a significant in-vitro increase (94%) in 142BR cell line proliferation at 20 microg/ml compared with the control. CONCLUSIONS: Compounds 1 and 2, which were isolated from the active chloroform fraction, have not previously been reported and showed a dose-dependent increase in proliferation of 142BR cells up to 3 microM; compounds 3, 4 and 5 had no effect on the 142BR cell line at the concentrations tested.

Profile of heavy metals in some medicinal plants from Ghana commonly used as components of herbal formulations.

The levels of some heavy metals in 27 medicinal plant species from Ghana were studied in order to evaluate their health implications. These plant species, especially those used in the treatment of diseases such as hypertension, diabetes and asthma may require long term usage. The metals were copper, zinc, iron, manganese, nickel and cadmium. Atomic Absorption Spectrophotometry (wet digestion) was used for the analyses, and content of metals per sample was expressed as percent µg/g. Daily total intake of these metals is discussed based on the recommended daily intake of the medicinal plants or their corresponding formulations. From the results of the study zinc, copper and cadmium were present in all the plant species examined. Manganese was present in all species except V. amygdalina. Iron was found in all except five species (82%), whilst nickel was (rather rare) detected in only eight (30%) of the plant species. Significant variations in metal content existed (P<0.05) among the medicinal plant species with respect to the heavy metals evaluated. The concentrations of copper, zinc, cadmium and manganese were within their respective maximum permissible daily levels. However, some species, especially Ocimum canum (8), Clausena anisata and Rauwolfia vomitoria had levels of iron higher than the maximum permissible level of 1000 µg/day and may require care to avoid iron toxicity. The results also highlighted the differences in contents of minerals in Lippia multiflora obtained from different locations in Ghana. The findings generally suggest that the use of these plant species for the management of diseases will not cause heavy metal toxicity and may be beneficial to the users in cases of micronutrient deficiency, as these metals were found to be present in readily bioavailable form.

Antibacterial, antioxidant and fibroblast growth stimulation of aqueous extracts of Ficus asperifolia Miq. and Gossypium arboreum L., wound-healing plants of Ghana.

AIM OF THE STUDY: Use of in vitro tests to search for relevant activities in bark of Ficus asperifolia Miq. and leaves of Gossypium arboreum L., used in Ghana for wound healing. MATERIALS AND METHODS: Aqueous extracts of the relevant parts of the two species were tested for antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Micrococcus flavus, Escherichia coli, Pseudomonas aeruginosa and resistant strains of Staphylococcus aureus SA1199B, RN4220 and XU212. The effects of the plants on fibroblast growth stimulation as well as antioxidant protective effect against hydrogen peroxide induced damage in the same cell line were also studied. RESULTS: The extracts of Gossypium arboreum and Ficus asperifolia had weak antibacterial action against all bacteria tested. The extracts of the two plants had significant (p<0.001) effects on the growth of human dermal fibroblast at 50 microg/ml and lower. They also significantly protected fibroblast cells against oxidative damage at doses up to 50 microg/ml. Gossypium arboreum leaf extract displayed the higher activity. CONCLUSION: The stimulatory effect on fibroblast growth and protection against peroxide-induced oxidative damage give some support to the traditional use of these two plants as wound-healing agents.

Estudio a fondo sobre todas as formas de violencia contra la mujer: informe del secretario general

Presente el estudio del informe del Sr. Kofi Annan, secretario general de las Naciones Unidas, acerca de la violencia contra la mujer. Trae temas como: derechos humanos, contexto y causas de la violencia, desigualdades de género y económicos y otros temas. Documento en formato PDF, requiere Acrobat Reader.