Effet antihyperglyc��miant des extraits de Boscia senegalensis (Pers.) Lam. ex Poiret et de Colocynthis vulgaris (L.) Schrad

Boscia senegalensis and Colocynthis vulgaris are medicinal plants widely used by Chadian population in the folk medicine to treat diabetes. The hydro alcoholic extracts of the seeds of Boscia senegalensis and the fruits of Colocynthis vulgaris have been valued on the albino rabbits whose hyperglyceamia has been induced by oral administration of D(+)-glucose. The seed extract of Boscia senegalensis, characterized by the presence of alkaloids, saponosids, tanins and mucilages, presented an optimal activity at the dose of 250mg/kg, while the fruits extract of Colocynthis vulgaris showed a maximal activity at the dose of 100mg/kg; its main constituents were alkaloids, tanins, terpenoides and mucilages.     

Validation, transfer and measurement uncertainty estimation of an HPLC-UV method for the quantification of artemisinin in hydro alcoholic extracts of Artemisia annua L

Malaria is the world's most important parasitic infection with 500 millions cases annually and almost 2 millions death per year. This disease is more present in Sub-Saharan Africa where 90% of the infections are found. Artemisinin and its semi synthetic derivatives (artemether, artesunate) have actually the most powerful activity on malaria, even in its complicated forms and resistance cases.Various methods have been proposed for detection and quantification of artemisinin in Artemisia annua L. by HPLC-UV, but the plant extracts used for this quantification were extracts obtained with organic solvents (toluene, petroleum ether, hexane). To be able to use crude A. annua extracts prepared at low cost to formulate antipaludic drugs, we chose the use of a mixture of water and ethanol as solvent of extraction, but no adequate analytical method for this kind of extracts is published.The main objectives of this work were first to develop an analytical method for artemisinin quantification in hydro alcoholic extracts of A. annua. Second, this method had to be thoroughly validated by the research and development laboratory and, third, the transfer of this method to the routine laboratory had to be demonstrated. The final aim was to compare the estimation of measurement uncertainty obtained during the method validation with validation standards to measurement uncertainty estimates obtained during the method transfer study with real samples.The method was validated following the accuracy profile methodology and was found to be accurate in the concentration range of 10.0-54.0 μg/ml with CV < 8%. Limit of detection and of quantification were 2.73 and 10.0 μg/ml, respectively. The method was then successfully transferred to a laboratory in Benin by showing that the quality of the results that it will generate during routine application of the method is sufficient. Finally, the measurement uncertainty of the method was estimated from the validation experiments as well as from the transfer study with authentic unspiked samples of A. annua. The comparison of these measurement uncertainty estimations showed that they were coherent. It confirmed thus that the estimation of measurement uncertainty from validation experiments predicts well the measurement uncertainty of real routine samples. This analytical method was thus shown to be convenient for routine analysis of hydro alcoholic extracts of A. annua in Benin.     

In vitro antiplasmodial activity of plants used in Benin in traditional medicine to treat malaria

Aim of the study

The aim of the study was to evaluate the in vitro antiplasmodial activity of crude extracts of 12 plant species traditionally used in Benin for the treatment of malaria in order to validate their use.

Materials and methods

For each species, dichloromethane, methanol and total aqueous extracts were tested. The antiplasmodial activity of extracts was evaluated using the measurement of the plasmodial lactate dehydrogenase activity on chloroquine-sensitive (3D7) and resistant (W2) strains of Plasmodium falciparum. The selectivity of the different extracts was evaluated using the MTT test on J774 macrophage-like murine cells and WI38 human normal fibroblasts.

Results

The best growth inhibition of both strains of Plasmodium falciparum was observed with the dichloromethane extracts of Acanthospermum hispidum DC. (Asteraceae) (IC₅₀ = 7.5 μg/ml on 3D7 and 4.8 μg/ml on W2), Keetia leucantha (K. Krause) Bridson (syn. Plectronia leucantha Krause) (Rubiaceae) leaves and twigs (IC₅₀ = 13.8 and 11.3 μg/ml on 3D7 and IC₅₀ = 26.5 and 15.8 μg/ml on W2, respectively), Carpolobia lutea G.Don. (Polygalaceae) (IC₅₀ = 19.4 μg/ml on 3D7 and 8.1 μg/ml on W2) and Strychnos spinosa Lam. (Loganiaceae) leaves (IC₅₀ = 15.6 μg/ml on 3D7 and 8.9 μg/ml on W2). All these extracts had a low cytotoxicity.

Conclusion

Our study gives some justifications for the traditional uses of some investigated plants.     

Effect of antenatal gymnastics on childbirth: a study on 50 sedentary women in the Republic of Benin during the second and third quarters of pregnancy

Antenatal gymnastics is a form of non-pharmacological childbirth preparation methods. It helps maintain rachidian statics and to relieve painful syndromes (joints, lumbar pains) during pregnancy. It also helps correct gravidic problems (leg cramps, back ache, weight increase, etc.). It is both a physical and psychological training method in accordance with the natural mechanisms of childbirth, implying personal involvement on the part of the parturient. Fifty (50) pregnant women of Benin (apart from pathological pregnancies) divided into two groups of 25 subjects (experimental group) and 25 subjects (control group) voluntarily took part in the study after having given their written assent. Each involved subject is paired with a counterpart of control following the same criteria: age of the gestante, parity, characteristic of the pelvis. Subjects aged from 17 to 42 trained twice a week with a minimum of 24 hours rest between two sessions; the training period intensity of exercise is limited to 60% of the maximum heart frequency and each training session lasts 45 min. The results show a lower number of caesareans and a significant lower number of untorn perineas in the experimental group. Apgar scores are also statistically higher in the children of trained women. This study confirms that sedentary pregnant women without any medical counter-indications (repetitive miscarriages, premature births, placenta praevia, cervical gaping, cardiac diseases, hypertension, narrow pelvis) can participate in antenatal gymnastics at a moderated rhythm, for their own benefit and that of their babies. Muscular force improvement (quality of the abdominal pushing), psychological control (absence of stress and panic), and good body form, can be noticed in trained mothers. However, abnormal presentation of baby, fetal suffering and the inherent characteristics of the pelvis (narrow) are factors requiring a caesarean. On the whole, antenatal gymnastics is an effective non-pharmacological means to avoid complications in childbirth. This method might be one of the solutions which could benefit pregnant women in Africa, considering its low cost compared to the exorbitant cost of medicines and hospitalisation.     

In vitro antitrypanosomal and antiplasmodial activities of crude extracts and essential oils of Ocimum gratissimum Linn from Benin and influence of vegetative stage

Ethnopharmacological relevance

Different parts of Ocimum gratissimum Linn are largely used in folk medicine for the treatment of many diseases, some of which related to parasitical infections as fevers and headaches. In order to validate their use and to clarify the plant part which possesses the best antiparasitic properties, we decided to evaluate the in vitro antiplasmodial and antitrypanosomal activities of essential oils and crude extracts from leaves, stems and seeds of Ocimum gratissimum as well as their cytotoxicity.

Materials and methods

The essential oils and ethanol crude extracts of leaves and stems of Ocimum gratissimum from Benin, were obtained in pre and full flowering stages. Seeds obtained only in full flowering stage, were also extracted. The oils were isolated by hydrodistillation and analyzed by GC/MS and GC/FID. Extracts and essential oils were tested in vitro against Trypanosoma brucei brucei and Plasmodium falciparum. Cytotoxicity was evaluated in vitro against Chinese Hamster Ovary (CHO) cells and the human non cancer fibroblast cell line (WI38) through MTT assay to evaluate the selectivity and toxicity was assessed against Artemia salina Leach.

Results

The essential oils and non-volatile crude extracts of Ocimum gratissimum were more active on Trypanosoma brucei brucei than on Plasmodium falciparum (3D7). This activity varies according to the vegetative stage (pre and full flowering) and the plant part (seeds, stems and leaves) extracted. The best growth inhibition of Trypanosoma brucei brucei was observed with ethanol crude extracts of leaves (IC₅₀=1.66±0.48 μg/mL) and seeds (IC₅₀=1.29±0.42 μg/mL) in full flowering stage with good selectivity (SI>10). The chemical composition of the essential oil from aerial parts (47 compounds), characterized by the presence as main constituents of p-cymene, thymol, γ-terpinene, β-myrcene and α-thujene, depends on the vegetative stage. The oil contained some minor compounds such as myrcene (IC₅₀=2.24±0.27 μg/mL), citronellal (IC₅₀=2.76±1.55 μg/mL), limonene (IC₅₀=4.24±2.27 μg/mL), with good antitrypanosomal activities. These oils and crude extracts were not toxic against Artemia salina Leach and had a low cytotoxicity except leaves and seeds ethanol extracts obtained in full flowering which showed toxicity against CHO and WI38 cells.

Conclusions

Our study shows that ethanol crude extracts of leaves and seeds of Ocimum gratissimum in full flowering stage can be a good source of antitrypanosomal agents. This is the first report about the relation between the plant part extracted, the vegetative stage of the plant, the antitrypanosomal and antiplasmodial activities and the cytotoxicity of essential oils and non-volatile extracts of Ocimum gratissimum from Benin.     

Potential of the bush mint, Hyptis suaveolens essential oil for personal protection against mosquito biting

We studied the potential of the essential oil extract from the bush mint, Hyptis suaveolens, for use against mosquito biting under both laboratory and field conditions. In the laboratory, the repellency of various concentrations (1-6%) of the essential oil was assessed against Anopheles gambiae, based on a 15-min landing and biting on treated forearms of volunteers. In the laboratory, the percentage of mosquitoes landing on the forearm was 42, 33, 23, 23, 9, and 2 for 1%, 2%, 3%, 4%, 5%, and 6% essential oil concentration, respectively; and 92 and 91 for the solvent (isopropanol) and untreated control, respectively. The percentage of mosquitoes taking a blood meal was 22, 12, 13, 12, 5, and 3 for 1%, 2%, 3%, 4%, 5%, and 6% essential oil, respectively; and 52 and 51 for the solvent and control, respectively. In the field, the 6% essential oil repelled all mosquitoes immediately postapplication; this activity declined to 75% after 5 h. The repellent action of the 8% essential oil concentration was higher, 97% after 5 h. Based on these data, the essential oil of H. suaveolens appears to be a good candidate for use in the integrated management of mosquito vectors of disease.     

Snail intermediate host/Schistosoma haematobium relationships from three transmission sites in Benin (West Africa)

The relationships between three strains of Schistosoma haematobium (Doh, Sô-Tchanhoué and Toho-Todougba; from Benin, West Africa) and their snail hosts were assessed by measurement of several life-history traits, including the infection rate; pre-patent period; cercarial production of each parasite strain; and growth, fecundity and survival of the host snails. Adaptations to its local snail host was found for the Toho-Todougba strain and included a short pre-patent period, a long patent period and production of more cercariae in its local snail host. In contrast, the life-history traits of the Doh and Sô-Tchanhoué strains indicated non-local adaptations, as some sympatric host–parasite combinations were not compatible, the highest infection rates occurred in the allopatric snail Bulinus wrighti, and the duration of cercarial production was short because of the high level of mortality of the snails. Furthermore, snail reproduction ceased following infection by each of the three parasite strains, and the life-history traits were not influenced by the miracidial dose.     

Toxicity and gastric tolerance of essential oils from Cymbopogon citratus, Ocimum gratissimum and Ocimum basilicum in Wistar rats

Oils of Cymbopogon citratus, Ocimum gratissimum and Ocimum basilicum are widely used for their medicinal properties, and as food flavours and perfumes. Recently in a study in West Africa, these oils have been recommended to combat Fusarium verticillioides and subsequent fumonisin contamination in stored maize, but their toxicological profile was not investigated. The current study was undertaken to provide data on acute and subacute toxicity as well as on gastric tolerance of these oils in rat. For this purpose, the oils were given by gavage to Wistar rats for 14 consecutive days. The animals were observed daily for their general behaviour and survival, and their visceral organs such as stomach and liver were taken after sacrifice for histological analyses. A dose-dependent effect of the tested oils was observed during the study. Applied at doses generally higher than 1500 mg/kg body weight, the oils caused significant functional damages to stomach and liver of rat. Unlike the other oils, administration of O. gratissimum oil did not result in adverse effects in rat liver at the tested doses. The no observed adverse effect level (NOAEL) of the tested oils has been established. The three tested oils can be considered as safe to human when applied on stored maize at recommended concentrations.     

Structure–activity relationship study of thiosemicarbazones on an African trypanosome: Trypanosoma brucei brucei

To explore the structure–activity relationships of thiosemicarbazones on African trypanosome: Trypanosoma brucei brucei, a series of thirty-five thiosemicarbazones (1–35) have been synthesized and characterized by their ¹H NMR, ¹³C NMR, and FT-IR spectra. All compounds were tested for trypanocidal activity using the method “Lilit alamar blue”. The comparison of trypanocidal power of thiosemicarbazones was performed considering their structures. This study that was done using acetophenone thiosemicarbazone (1) as basic model, showed that: (a) the presence of lipophilic substituents in para position on benzene ring, (b) substitution of benzene ring and (c) substitution of hydrogen of thioamide function by a phenyl, strongly influence trypanocidal activity. The various modifications to basic structure (1) allowed the synthesis of 1-(4-chlorophenyl) ethylidene-4-phenyl-thiosemicarbazide (34). With a trypanocidal activity of 3.97 μM, this compound is the most active of the series.