Antiplasmodial Test Research Articles

In vivo Antiplasmodial Assessment of Phyllanthus odontadenius against Plasmodium berghei in NMRI Mice

Aims: Phyllanthus odontadenius is one of the genus Phyllanthus species, used for number diseases treatment including malaria. Malaria today poses a real public health problem for more than hundred countries, afflicted millions people and killed an estimated 405 000 in the World. The chemoresistance of Plasmodium falciparum to modern antimalarials which are either expensive, toxic or ineffective leads to the search for new antimalarials at lower cost, non-toxic and effective within plant biodiversity. Given the interest in various P. odontadenius crude extracts in vitro on P. falciparum, an in vivo study seems necessary in order to judge the extracts effectiveness of this plant. This study therefore aims to find justification for P. odontadenius secondary metabolites antiplasmodial activity which would have been revealed in the three samples from three different province sites. Study Design: Plants P. odontadenius samples from three province harvested and dried separately, plant phytochemistry screening realized, extracts preparation for antiplasmodial test, mice parasitization with P. berghei strain, orally administration drug; Parasiteamia determination using an immersion microscope. Place and Duration of Study: Department of Radiobiology, Applied Microbiology section, General Atomic Energy Commission, Regional Nuclear Studies Center of Kinshasa. MPI and pharmacognosy laboratories in the National Biomedical Research Institute (INRB). This work took place over the period from October 11, 2020 to March 12, 2021. Methodology: Phytochemical screening P. odontadenius samples was previously determined with the chemical reagent reactions and TLC. Then, P. odontadenius methanol extracts from aerial parts harvested in three sites (Kinshasa, Kasangulu and Kwango-bridge) were administered to test mice (12.5 mg/kg and 25 mg//kg bw) after infected mice with the Plasmodium berghei strain. DMSO 10% and quinine 10 mg/kg bw were also used as controls for comparison with the samples of P. odontadenius extracts. After 5 days, parasitemia of each test and controls mice was determined. Percent of parasitemia, parasite density and percent of inhibition were calculated. Finally, the effect dose 50 of each P. odontadenius specimen was finally determined. Results: Parasiteamia rates of negative control (DMSO 10%) was high (69.98±15.03%) comparing to positive control (27.43±11.46%) and tested mice with P. odontadenius extracts (12.5 and 25 mg/kg bw) which percent’s varied from 24.66±15.84% to 59.01±22.44%. Negative control presented high parasite density with 11,342 (±2,436) comparing to the positive control (4,447±1,857) and all P. odontadenius methanol extracts which varied from 3,995±2,343 for 25 mg/kg bw to 9,570±3,319 for 12.5 mg/kg bw. Parasiteamia reduction rates followed inversely parasite density, thus, Po3 25 mg/kg bw had high parasiteamia reduction rate (65.23%) comparing to positive control with 61.32% and to P. odontadenius methanol extracts. Po3 presented 2.44 mg/kg bw as effect dose 50 comparing to Po1 (2.93 mg/kg bw) and Po2 (2.68 mg/kg bw). Males mice were highly affected to P. berghei than the females. Conclusion: This study revealed that all specimens of P. odontadenius had good in vivo antiplasmodial activities on P. berghei. All P. odontadenius extracts showed good parasitaemia inhibition compared to negative control, but P. odontadenius from Kwango-brigde (Po3) presented good behavior concerning in vivo antiplasmodial activity in comparison to P. odontadenius from Kasangulu (Po2) and that from Kinshasa (Po1). However, further studies are necessary on the in vivo toxicity of the plant and on the medicinal form that could be applied.

The ethnopharmacological study of plant drugs used traditionally in Djibouti for malaria treatment

Ethnopharmacological relevanceDjibouti was a country where malaria has been endemic for centuries. The local population use the plants as repellents or first aid for uncomplicated malaria. Aim of the studyThe aim was, for the first time, to collect and identify plants used by the local population to treat malaria and select the most interesting plants (those that are more commontly used, more available, and have fewer studies). These plants were evaluated for their antiplasmodial activity as well as their cytotoxicity on human cell lines for the most active ones. Materials and methodsA semi-structured questionnaire was developed for this study to collect information about the use and identity of botanical drugs used to treat malaria. The use-reports (percentage) of each plant were recorded to determine their use importance. Also, the availability status of the plants was assessed; and those in critical condition were discarded excluded from further study. Fifteen plants, out of the 41 listed, were extracted with hydro alcohol, ethyl acetate, and dichloromethane for biological testing. Chloroquine-resistant strain FcB-1 of P. falciparum and a human diploid embryonic lung cell line were used for the antiplasmodial test, and to assess the cytotoxicity for human cells respectively. Preliminary analysis of extract constituents was carried out using thin layer chromatography (TLC). ResultsThis study identifies 41 plant taxa belonging to 32 families and records their use against malaria. Balanites rodunfolia, belonging to the Zygophyllaceae family, was the most commonly used plant, representing 44 % of use-reports. It was followed by Cadaba rodunfolia (15 %) from the Capparaceae family, and then the three species of Aloe: Aloe djiboutiensis (8.2 %), Aloe ericahenriettae (3.4 %), and Aloe rigens (3.4 %) from the Asphodelaceae family. The leaves are the most commonly used part of the plants to treat malaria, accounting for 76 % of usage. The preparation methods were decoction (52 %), maceration (29 %), and boiling (19 %). The administration routes were by oral (80 %), inhalation 19 %), and bathing (1 %). The best antiplasmodial activities were observed in the dichloromethane extracts of Cymbopogon commutatus and the ethyl acetate extracts of Aloe rigens and Terminalia brownii, with IC50 values of 9.8, 5, and 7.5 μg/mL, respectively. Their toxicity/activity levels were very favorable with selectivity indices of 5.6, 8.1, and 11.8 for C. commutatus, A. rigens, and T. Brownii, respectively. ConclusionForty-one species of botanical drugs were listed as being used to treat malaria in Djibouti. All fifteen selected species showed antiplasmodial activity (IC50 < 50 μg/mL). This work will help guide the valorization of botanical drugs used to treat malaria in Djibouti.

West African medicinal plants: a review of their antimalarial activity

Nowadays, the use of medicinal plants in the fight against malaria must be based on scientific results of safety and quality. However, reviews of the antiplasmodial activities of plants in West Africa in recent years are rare. This study analyzes scientific publications from 2010 to 2021 on plants traditionally used in antimalarial treatments in West Africa. A systematic search was carried out in the PubMed and google scholar databases using the following keywords: Malaria, Antiplasmodial activity, extract, medicinal plant, West Africa; for articles published from 2010 to 2021. These articles concern ethnobotanical studies, antiplasmodial tests, isolated molecules and toxicity tests. A total of 8 West African countries were explored and 54 papers from 2010 to 2021 were selected with 78 plants studied. Nigeria and Burkina Faso recorded more work with 28 and 7 papers respectively and studied more plants with 31, and 16 respectively. The most active extracts for in vitro tests are found in Nigeria with ethanolic extracts of Phyllanthus amarus and Ipomoea purpurea with respectively an Inhibitory Concentratin of 0.05 μg /mL and 0.06 μg / mL. The most active extract in vivo is found in Nigeria with the methanolic fraction of Parkia biglobosa with a 100% suppression rate at a dose of 100 mg/kg/Day. It is clear that the traditional West African pharmacopoeia is a potential source of effective phytomedicines for the management of malaria.

Comparative Assessment of the Antiplasmodial Activity and Acute Toxicity of Schumanniophyton magnificum Good &amp; Halle (Rubiaceae) Leaves, Trunk and Roots Methanolic Extracts

Background: Despite the various medicinal applications of Schumanniophyton magnificum, no comparative data are available to determine the part with best antiplasmodial activity and to guarantee its safety. This work deals with the evaluation of the antiplasmodial activity of methanolic extracts of S. magnificum organs as well as the study of acute toxicity of the most active part.&#x0D; Materials and methods: The methanolic extracts of the leaves, stem bark, trunk wood, root wood and root bark, were obtained by maceration with 96% methanol. Phytochemical screening, based on precipitation and coloring reactions, to highlight the chemical groups present in the plant. The antiplasmodial test was ran against chloroquine sensitive Plasmodium falciparum 3D7 strains while the study of the in vivo acute toxicity was conducted according to guideline 423 of the OECD protocol at a fixed dose on Wistar rats.&#x0D; Results: Phytochemical screening of the methanol extracts of these plants organs revealed the presence of alkaloids, triterpenoids, saponins, anthraquinones, polyphenols, coumarins anthocyanins and flavonoids. The antiplasmodial activity showed that only the leaves exhibited a moderate antiplasmodial effect with IC50 of 30.77 µg/ml. An oral administration of the methanolic extract of the leaves did not induce an abnormal variation of the physiological parameters in female Wistar laboratory rats, at non-toxic doses of 50, 300 and 2000 mg/kg body weight for 14 days.&#x0D; Conclusion: The leaves of S. magnificum exhibited the best antimalarial activity and were non toxic, thus justifying the use of the plant in the treatment of malaria in traditional pharmacopoeia.&#x0D; Keywords: Phytochemical screening, Antiplasmodial activity, Acute toxicity, Schumanniophyton magnificum, Medicinal plant.

Comparative Anti-plasmodial and Cytotoxic Effect of the Methanolic Extracts of Leaves, Stem Bark and Roots of Annickia affinis

Aims: This study aimed to evaluate and compare the in vitro anti-plasmodial and cytotoxic effect of the methanolic extracts from leaves, stem bark and roots of Annickia affinis.&#x0D; Study Design: This is an experimental study.&#x0D; Place and Duration of Study: The work was conducted at the Pharmacochemistry and Natural Substances Laboratories of the Faculty of Medicine and Pharmaceutical Sciences, University of Douala for the extraction and at the Biomedical Chemistry Research Center of Rhodes University in South Africa for the antimalarial and cytotoxic essay. All the experiments were carried out from the 15th October 2019 to the 31th July 2020.&#x0D; Methodology: The anti-plasmodial test was performed on Plasmodium falciparum sensitive strains 3D7 while cytotoxicity was evaluated on the HeLa cell line.&#x0D; Results: The anti-plasmodial tests revealed that the roots and the stem bark exhibited a moderate anti-plasmodial effect with IC50 of 19.7 ± 0.8 and 12.1 ± 0.8 µg/ml respectively. The anti-plasmodial effects of the leaves were classified as low (33.7 ± 1.9 µg/ml). At up to 50 µg/ml, all the extracts showed a high rate of survival among the HeLa cells. No effect was observed with the leaf extracts (100% of survival).&#x0D; Conclusion: This is the first report on the cytotoxic study and comparative anti-plasmodial effect of Annikia affinis. It highlights the potential of Annickia affinis as an important source of anti-plasmodial drugs with less cytotoxic in vitro. In agreement with the use in traditional medicine, the stem bark was more active than wood, while leaves showed low activity.

In vitro antiplasmodial activity and cytotoxicity of extracts and chromatographic fractions of twigs from Pappea capensis EckI & Zeyh. (Sapindaceae)

Ethnopharmacological relevanceThe Vha-Venda people of South Africa use Pappea capensis EckI & Zeyh. (Sapindaceae) twigs to treat malaria and its related symptoms. Aim of studyThe main aim of this study was to evaluate the antiplasmodial and cytotoxic activity of P. capensis extracts and chromatographic fractions. Spectroscopy analysis was conducted using 1H NMR and GC-MS to tentatively identify the major classes of compounds and phytoconstituents that can be attributed to the observed antiplasmodial bioactivity. Materials and methodsPappea capensis twigs were dried and then ground to fine powder. A solvent mixture of dichloromethane: methanol: water (1:0.5:0.5, v/v) was used to extract. The polar extract was separated from the non-polar. The organic extract was dried to yield a DCM (I = 60 g) extract. The methanol in the aqueous extract was evaporated using a rotary vapour and the remaining water freeze dried to yield a water extract (II = 287 g). Extract I was further partitioned using a solvent mixture of DCM: MeOH (1:1, v/v), separated and concentrated under vacuum to yield dichloromethane (III = 40 g) and methanol (IV = 15 g) extracts. A water-based decoction (V = 10 g) was also prepared to establish the clinical relevance of the preparation administered by Vha-Venda people in South Africa. Extracts II, III and IV were further subjected to silica column chromatography, eluting with a series of different solvents with increasing polarity to yield a total of 25 fractions (A – Y). In vitro antiplasmodial tests on Plasmodium falciparum (NF54) and cytotoxicity screens on mammalian L-6 rat skeletal myoblast cells were performed on all extracts and fractions. Selectivity indices (SI) were also computed for all tested extracts and fractions which were further subjected to 1H NMR spectroscopy and GC-MS analysis for the identification of the major classes of compounds present in the extracts. ResultsFrom the assayed extracts, only extract I (IC50 = 2.93 μg/ml; SI = 14), III (IC50 = 2.59 μg/ml; SI = 21) and IV (IC50 = 3.56 μg/ml; SI = 13) demonstrated the best antiplasmodial activity and selectivity. Of all assayed fractions, only N (0.6 μg/ml; SI = 91), D (0.85 μg/ml; SI = 37) and E (0.91 μg/ml; SI = 30) depicted the best antiplasmodial activity and selectivity. The 1H NMR analysis of the extracts and fractions identified the prominent class of constituents to be aliphatic based which was tentatively identified as terpenoids. When further GC-MS analysis was conducted, the presence of lupin-3-one, lupeol acetate, α-amyrin, and β-amyrin phytoconstituents were tentatively confirmed. These constituents are triterpenoids with established antiplasmodial activity which can be tentatively attributed to the bioactivity observed in P. capensis twigs. ConclusionThe study validates the ethnomedicinal use of P. capensis for malaria treatment. It demonstrated the potential of discovering novel antiplasmodial constituents that could serve as drug hits through dereplication approaches where known compounds with established antimalarial activity can be bypassed to focus on the unknown.

Evaluation of the in vivo anti-plasmodial activity of ethyl-acetate extract of Cassia occidentalis leaf in Plasmodium berghei – infected mice

Background: Malaria is undoubtedly one of the world’s deadliest infectious diseases. It is a widespread disease that continues to be associated with considerable morbidity and mortality with a significant social and economic impact in developing countries. This study was aimed at evaluating the therapeutic effect of the ethyl acetate extract of Cassia occidentalis in Plasmodium berghei infected mice. Methods: Qualitative phytochemical screening was carried out, using standard methods. Acute oral toxicity (LD50), as well as both suppressive and curative antiplasmodial test, was carried out. Results: Flavonoids, saponins, steroids, tannins and phenolic compounds were found to be present in the extract. The median lethal dose (LD50) was calculated to be ≥ 5000mg kg-1 body weight. Percentage suppression of parasitaemia was calculated to be 37.84, 42.84, 77.88, 83.06 and 0.00 %, with mean survival period (in days) of 6.00±0.58, 12.33±1.45, 26.33±2.03, 29.00±1.00 and 4.33±0.88 for groups treated with 100, 200, 400 mg kg-1 bodyweight of the extract, 5 mg kg-1 body weight of chloroquine and negative control group respectively. The percentage inhibition of parasitaemia in the curative test was calculated to be 42.38, 47.67, 79.17, 81.99 and 0.00 %, with mean survival period (in days) calculated to be 7.67±1.45, 15.00± 1.73, 25.33±1.86, 29.00±1.00 and 5.00± 1.53 for groups treated with 100, 200, 400 mg kg-1 bodyweight of the extract, 5 mg kg-1 body weight of chloroquine and the negative control group respectively. It was observed that there was a significant (p&lt;0.05) increase in percentage packed cell volume (PCV) in the treated groups compared to the untreated group. Conclusion: The result of this study showed that ethyl acetate extract of Cassia occidentalis leaf possess appreciable antiplasmodial effect and justifies its traditional use as an herbal remedy against malaria and may, therefore; offer a potential drug lead for the development of a safe, effective and affordable antimalarial.

Growth, secondary metabolite production, and in vitro antiplasmodial activity of Sonchus arvensis L. callus under dolomite [CaMg(CO3)2] treatment.

Malaria is still a global health problem. Plasmodium is a single-cell protozoan parasite that causes malaria and is transmitted to humans through the female Anopheles mosquito. The previous study showed that Sonchus arvensis L. callus has antiplasmodial activity. Several treatments are needed for callus quality improvement for antimalarial compound production. This study aimed to examine the effect of dolomite [CaMg(CO3)2] on growth (morpho-anatomical structure and biomass), secondary metabolite production, and in vitro antiplasmodial activity of S. arvensis L. callus. In this study, leaf explants were grown in Murashige and Skoog medium with a combination of 2,4-dichlorophenoxyacetic acid (2,4-D, one mg/L) and 6-benzyl amino purine (BAP, 0.5 mg/L) with dolomite (50, 75, 100, 150, and 200 mg/L). The 21 days callus ethanolic and methanolic extract were analyzed by gas chromatography-mass spectrometry (GC-MS) and thin-layer chromatography (TLC). The antiplasmodial test was performed on a blood culture infected with Plasmodium falciparum strain 3D7 using the Rieckmann method. The results showed that dolomite significantly affected callus growth, metabolite profile, and in vitro antiplasmodial activity. Dolomite (150 mg/L) showed the highest biomass (0.590 ± 0.136 g fresh weight and 0.074 ± 0.008 g dry weight). GC-MS analysis detected four compounds from callus ethanolic extract. Pelargonic acid, decanoic acid, and hexadecanoic acid were major compounds. One new terpenoid compound is based on TLC analysis. S. arvensis L. callus has antiplasmodial activity with the IC50 value of 5.037 μg/mL. It was three times lower than leaf methanolic extract and five times lower than leaf ethanolic extract.

Microwave assisted extraction of antiplasmodial and antioxidant limonoids from Trichilia roka (chiov) Meliaceae)

Trichilia roka (Chiov) is a medicinal plant from Meliaceae family. It is recognized in traditional medicine for its innumerable therapeutic properties. Limonoids, the main constituents of the root bark of this plant, is known for its antioxidant and antiplasmodial activities. To obtain an improved yield of these bioactive compounds from T. roka and reduce extraction time, solvent and energy required, it is of utmost importance to adopt innovative approaches such as microwave-assisted extraction. Microwave was attempted, as compared with the conventional mechanical agitation method to extract bioactive limonoids and quantify them through colorimetric quantification method using 4(dimethyl amino) benzaldehyde (DMAB). The antiplasmodial activity was evaluated against the intraerythrocytic stages of cultured Plasmodium falciparum using a phenotypic approach, and the antioxidant property was evaluated in vitro using DPPH radical-scavenging and β-carotene-linoleate model systems respectively. Three limonoids and were isolated from Trichilia roka (Chiov) root bark labelled RA, RO and RY. The microwave extraction yields were 115.895 mgRUBE/gDW for limonoids. The isolated compounds exhibited good antioxidant activities than crude extracts with IC50 values of 2.59 10-3 , 2.26 10-3 and 1.79 10-3 mg/mL respectively compared to crude extract IC50 values of 2. 10-2 mg/mL. The in vivo antiplasmodial test of the hydromethanolic microwave extracts showed during the five treatment days the decreasing of the parasitaemia for doses 100 mg/kg, 200 mg/kg and 500 mg/kg with inhibition percentages of 82.75, 84.84 and 87.8 respectively.

Antiplasmodial activity of Ethanolic extract of Cassia spectabilis DC leaf and its inhibition effect in Heme detoxification

BackgroundIn previous studies, Cassia spectabilis DC leaf has shown a good antiplasmodial activity. Therefore, this study is a follow-up study of the extract of leaf of C. spectabilis DC on its in vitro and in vivo antiplasmodial activity and mechanism as an antimalarial.MethodsThe extract was fractionated, sub-fractionated and isolated to obtain the purified compound. In vitro antiplasmodial activity test against Plasmodium falciparum to find out the active compound. In vivo test against P. berghei ANKA-infected mice was conducted to determine prophylactic activity and antiplasmodial activity either alone or in combination with artesunate. The inhibition of heme detoxification test as one of the antimalarial mechanisms was carried out using the Basilico method.ResultsThe results showed that active antimalarial compound isolated from C. spectabilis DC leaf had a structural pattern that was identical to (−)-7-hydroxycassine. Prophylactic test of 90% ethanolic extract of C. spectabilis DC leaf alone against P. berghei ANKA-infected mice obtained the highest percentage inhibition was 68.61%, while positive control (doxycycline 13 mg/kg) was 73.54%. In combination with artesunate, 150 mg/kg three times a day of C. spectabilis DC (D0-D2) + artesunate (D2) was better than the standard combination of amodiaquine + artesunate where the inhibition percentages were 99.18 and 92.88%, respectively. The IC50 of the extract for the inhibitory activity of heme detoxification was 0.375 mg/ml which was better than chloroquine diphosphate (0.682 mg/ml).ConclusionC. spectabilis DC leaf possessed potent antiplasmodial activity and may offer a potential agent for effective and affordable antimalarial phytomedicine.

Antiplasmodial Activity of The Low Molecular Weight Compounds from Streptomyces sp. GMR22

Low molecular weight (LMW) antiplasmodial compounds isolated from bacteria, particularly Streptomyces have not been widely reported. This study aimed to identify LMW compounds from Streptomyces sp. GMR22 as antiplasmodial. Isolation of the LMW compounds from the supernatant of fermentation culture using solvent of n-hexane:ethylacetate (EtOAc) (85:15v/v)and identified using gas chromatography-mass spectrometry (GC-MS). Antiplasmodial assay of n-hexane:EtOAc extract was carried out in vitro against P. falciparum (3D7). Parasitemia percentage obtained through microscopic observations and 50% inhibitory concentration (IC50) obtained through probit analysis. The antiplasmodial confirmation test was done by flow cytometry using SYBR Green I for Plasmodium DNA and anti-human CD235a for erythrocytes. The LMW compounds were investigated using SwissADME for drug-likeness. n-Hexane:EtOAc extract contained 21 LMW compounds from alcohol, hydrocarbon, ester, aromatic/diester, diester, fatty acid, and triester classes, which possessed moderate antiplasmodial activity with an IC50 value of 38.61 ± 19.06 µg/mL. Confirmation by flow cytometry analysis showed that the extract at 50 µg/mL exhibited antiplasmodial activity based on a decreased Plasmodium DNA intensity as compared to the control group. The result of drug-likeness screening obtained that 3 LMW compounds were drug-likeness, namely phenylethyl alcohol, ethyl citrate, and di-n-butyl phthalate. Streptomyces sp. GMR22 produced LMW compounds as antiplasmodial, and further study was needed for identification of antiplasmodial active compounds.

Biological Efficacy and Toxicological Evaluation of Ethanolic Extract of Cassia nodosa Buch.-Ham. (Leguminosae)

Medicinal plants are used worldwide to treat various diseases. However, scientific validation of the therapeutic potential of most medicinal plants is absent, and little to no information on their potential toxicity exists. It is therefore important to assess the efficacy and toxicity of various medicinal plants commonly used in traditional herbal medicine. The main goal of this study was to evaluate the efficacy and toxicity of the ethanolic extract of Cassia nodosa, a plant used in the treatment of fever, cold, gastric pain, diabetes mellitus, and malaria in Ghana. The stem bark extract of Cassia nodosa was obtained by cold maceration. Following phytochemical examination via standard procedures, the extract was screened for antiplasmodial, anti-inflammatory, and antioxidant activities. The extract was also evaluated for acute oral toxicity and its toxicity to the liver and kidney of rats. Phytochemicals present in the extract included sterols, alkaloids, tannins, glycosides, and coumarins. In the in vitro antiplasmodial activity test, the Cassia nodosa extract showed moderate antiplasmodial activity with an IC50 value of 23.98 ± 2.07 µg/ml. An ED50 value of 8.52 ± 2.83 mg/kg in the anti-inflammatory test indicated that the Cassia nodosa extract administered orally was very active in reducing inflammation. The total antioxidant capacity of the extract was determined to be 48.49 ± 5.24 gAAE/100 g with the total phenolic content being 28.48 ± 3.13 (gGAE/100 g) extract. In the DPPH radical scavenging assay, the concentration required for 50% inhibition of DPPH radical was found to be 78.71 ± 6.84 µg/mL. There was a gradual increase in the body weight of rats in both treated and control groups during the acute toxicity test. Clinical biochemical tests did not show any significant differences between control and treated groups of animals in terms of liver and renal functions, 14 days after administration of Cassia nodosa extracts. Taken together, these results provide scientific validation for the use of the plant in traditional medicine.

Synthesis of Novel 1,2,3-Triazole Derivatives of Isocoumarins and 3,4-Dihydroisocoumarin with Potential Antiplasmodial Activity In Vitro.

Malaria greatly affects the world health, having caused more than 228 million cases only in 2018. The emergence of drug resistance is one of the main problems in its treatment, demonstrating the need for the development of new antimalarial drugs. Synthesis and in vitro antiplasmodial evaluation of triazole compounds derived from isocoumarins and a 3,4-dihydroisocoumarin. The compounds were synthesized in 4 to 6-step reactions with the formation of the triazole ring via the Copper(I)-catalyzed 1,3-dipolar cycloaddition between isocoumarin or 3,4- dihydroisocoumarin azides and terminal alkynes. This key reaction provided compounds with an unprecedented connection of isocoumarin or 3,4-dihydroisocoumarin and the 1,2,3-triazole ring. The products were tested for their antiplasmodial activity against a Plasmodium falciparum chloroquine resistant and sensitive strains (W2 and 3D7, respectively). Thirty-one substances were efficiently obtained by the proposed routes with an overall yield of 25-53%. The active substances in the antiplasmodial test displayed IC50 values ranging from 0.68-2.89 μM and 0.85-2.07 μM against W2 and 3D7 strains, respectively. This study demonstrated the great potential of isocoumarin or 3,4-dihydroisocoumarin derivatives because practically all the tested substances were active against Plasmodium falciparum.

α-VINIFERIN as a potential antidiabetic and antiplasmodial extracted from Dipterocarpus littoralis

Over the past few decades, complementary medicine therapy using medicinal plants have been developed in healthcare. Phytochemical studies about medicinal plants have been conducted to verify their potency as medicinal remedies in modern therapeutics. Dipterocarpus littoralis commonly known as Meranti Jawa in Indonesia is traditionally used to treat diseases such as diarrhea, diabetic and malaria. This study aimed to isolate bioactive compounds from D. littoralis using bioguided fractionation method. The bioactivity measured were antioxidant, antidiabetic, and antiplasmodial activity. Alpha-glucosidase and alpha-amylase assays were applied to estimate the in vitro antidiabetic activity of D. littoralis. The antioxidant activities were determined by using the free radical scavenging assays 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-2″-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Analysis of total flavonoid and phenolic contents were expressed as Quercetin Equivalent (QE) and Gallic Acid Equivalent (GAE), respectively. The in vitro antiplasmodial activity test of methanol extract of D. littoralis was also conducted against Plasmodium falciparum strain 3D7. Purification of the ethyl acetate fraction of the methanol extract of D. littoralis resulted in an oligostilbenes namely α-viniferin (1). The structure of the α-viniferin was characterized by comprehensive spectral analysis including IR, 1D and 2D NMR, and in comparison with the literature data. Compound 1 showed an alpha-glucosidase and alpha-amylase inhibitory activity with IC50 values of 256.17 and 212.79 μg/mL, respectively. The in vitro antiplasmodial activity test against Plasmodium falciparum strain 3D7 at a concentration of 100 μg/mL revealed a strong antiplasmodial inhibitory activity with IC50 value of 2.76 μg/mL. Our findings indicated that α-viniferin (1) which is isolated from D. littoralis extract could be regarded as potential antidiabetic and antiplasmodial resources in the future.

CALLUS INDUCTION OF Sonchus arvensis L. AND ITS ANTIPLASMODIAL ACTIVITY.

Background:Malaria is a global health problem that requires urgent need for new drugs. Tempuyung (Sonchus arvensis L.) possesses many potential medicinal compounds. As the plant is originally found wild, it is important to reproduce its secondary metabolites by tissue culture. The objectives of this study were to look for effective methods to induce callus from leaf explants of Sonchus arvensis L. and to test its in vitro antiplasmodial activity.Materials and Methods:The leaves and petioles of the plant were cultured on Murashige and Skoog (MS) solid medium supplemented with indole acetic-3-acid (IAA), indole-3-butyric acid (IBA), naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and benzyl amino purine (BAP), in light and dark incubations. The best results obtained from callus induction were then treated by with several concentrations of sucrose (1- 5%). The best results from callus induction were then extracted with methanol for antiplasmodial test by Trager and Jensen’s method. It was also tested against 3D7 strain of Plasmodium falciparum.Results:The combination of 1mg/L 2,4-D and 0.5 mg/L BAP in dark incubation was the best treatment for callus induction of tempuyung. It produced the best quality of callus and the shortest period for callusing. Sucrose treatment had various effects on leaves callusing, but had no effect on petioles callusing, whereby 4% sucrose was the best treatment for leaves callusing in dark incubation. The methanol extract of the best callus had anti-plasmodial activity with IC50=0.343 µg/mL.Conclusion:Methanol extract of tempuyung callus shows potential as an antimalarial drug but more studies would be required.

A methodology for the isolation of marine microorganisms tolerant to hexavalent chrome for its bioreduction potential in wastewater

Over the past few decades, complementary medicine therapy using medicinal plants have been developed in healthcare. Phytochemical studies about medicinal plants have been conducted to verify their potency as medicinal remedies in modern therapeutics. Dipterocarpus littoralis commonly known as Meranti Jawa in Indonesia is traditionally used to treat diseases such as diarrhea, diabetic and malaria. This study aimed to isolate bioactive compounds from D. littoralis using bioguided fractionation method. The bioactivity measured were antioxidant, antidiabetic, and antiplasmodial activity. Alpha-glucosidase and alpha-amylase assays were applied to estimate the in vitro antidiabetic activity of D. littoralis. The antioxidant activities were determined by using the free radical scavenging assays 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-2″-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Analysis of total flavonoid and phenolic contents were expressed as Quercetin Equivalent (QE) and Gallic Acid Equivalent (GAE), respectively. The in vitro antiplasmodial activity test of methanol extract of D. littoralis was also conducted against Plasmodium falciparum strain 3D7. Purification of the ethyl acetate fraction of the methanol extract of D. littoralis resulted in an oligostilbenes namely α-viniferin (1). The structure of the α-viniferin was characterized by comprehensive spectral analysis including IR, 1D and 2D NMR, and in comparison with the literature data. Compound 1 showed an alpha-glucosidase and alpha-amylase inhibitory activity with IC50 values of 256.17 and 212.79 μg/mL, respectively. The in vitro antiplasmodial activity test against Plasmodium falciparum strain 3D7 at a concentration of 100 μg/mL revealed a strong antiplasmodial inhibitory activity with IC50 value of 2.76 μg/mL. Our findings indicated that α-viniferin (1) which is isolated from D. littoralis extract could be regarded as potential antidiabetic and antiplasmodial resources in the future.

Flavonoid Analysis and Antioxidant Activities of the Bryonia alba L. Aerial Parts.

Bryonia alba L. is the only Bryonia species found in Romanian flora, being known as a remedy for inflammatory pathologies or for its hepatoprotective and adaptogen activities. The present investigation studied the flavonoid composition and antioxidant activities of the aerial parts of this species. Flavonoid profile was evaluated by HPLC coupled with Diode Array Detection (DAD), while antioxidant capacity was assessed by various methods, testing different antioxidant mechanisms: DPPH (2,2-diphenyl-1-picrylhydrazyl), CUPRAC (cupric reducing antioxidant capacity), FRAP (ferric reducing ability of plasma), TEAC (Trolox equivalent antioxidant capacity), EPR (electron paramagnetic resonance method) and SNPAC (silver nanoparticles antioxidant capacity). Cytotoxicity was tested on human cancerous and healthy cell lines. Anti-plasmodial tests were performed on two strains of Plasmodium falciparum. Whole organism toxicity was assessed on zebrafish larvae. The HPLC-DAD analysis proved the presence of lutonarin, saponarin, isoorientin, and isovitexin as the major flavonoids in the composition of tested samples. Significant results were obtained for all antioxidant capacity assays. The cytotoxicity tests proved the absence of cellular and parasitic toxicity and these results were confirmed by the lack of toxicity on the zebrafish larvae model. This study proves a promising potential of the aerial parts of Bryonia alba L. as antioxidant agents.

THE EFFECT OF MALACCA LEAVES (Phyllantus emblica) ETHANOLIC EXTRACT ON Plasmodium falciparum GROWTH IN VITRO

The aim of this research was to find out in vitro antiplasmodium activity of Malacca leaves (Phyllantus emblica) ethanolic extract against Plasmodium falciparum growth. In this study, Plasmodium culture contained 5% parasitemia in ring stage was cultured using candle jar method and antiplasmodial activity test was carried out using microculture. The treatments were divided into 7 groups with four repetitions. K1 as negative control group was given Roswell Park Memorial Institute (RPMI), while K2 as positive control group was given artesdiaquine. Groups K3, K4, K5, K6, and K7 group was added with 100 µg/mL, 75 µg/mL, 50 µg/mL, 25 µg/mL, and 5 µg/mL of Malacca leaves ethanolic extract, respectively. Antiplasmodial activity was determined by inhibition concentration of 50% parasite growth (IC50). The data were analyzed using ANOVA and followed by Duncan test. The average of parasitemia level in group K1, K2, K3, K4, K5, K6, and K7 were 55.25±15.62, 8.50±2.52, 8.50±3.00, 9.25±0.95, 9.00±2.70, 9.79±2.06, and 10.75±2.22, respectively. The average of inhibition percentage in group K1, K2; K3; K4; K5; K6; and K7 were 0.00±0.00%, 84.62±4.55%; 84.62±5.43%; 83.26±1.73%; 83.71±4,90%; 82.35±3,73%; and 80.54±6.83%, respectively (P0.01). The results showed that the administration of malacca leaves ethanolic extract significantly affect (P0.01) the inhibition of Plasmodium growth as compared to group K1 (negative control). Probit analysis reveals the IC50 value was 3.889 µg/mL. In conclusion, all doses of malacca leaves ethanolic extract used in this study was able to inhibit Plasmodium falciparum growth with IC50 value was 3.889 µg/mL.

Improvement of Secondary Metabolites from Phyllanthus odontadenius against Malaria by Mutagenesis

Aims: Majority of deaths in children aged under 5 years are due to Plasmodium falciparum malaria. Malaria deaths in children decreased but malaria remains a major killer of children, taking the life of a child every 2 minutes. This study aims to investigate the increasing of the in vitro antiplasmodial activities by mutagenesis techniques using gamma-rays (Cs-137) or sodium azide (NaN3) as utagens. It will allow the importance of mutagenesis use as tools for improvement of secondary metabolites against malaria parasites using chemical or physical mutagens. Study Design: Laboratory experiment tests : identification of plant material, immersion of seeds in SA (sodium azide) solutions or irradiation by Gamma-rays (Cs-137) of P. odontadenius seeds for improvement of secondary metabolites against malaria parasites, in vitro culture of seeds followed by the in situ culturing of plantlets for obtaining material of study, phytochemical screening of Phyllanthus odontadenius aerial parts to determine the change of compounds in comparison to controls, in vitro antiplasmodial tests for the determination of SA concentrations or those of gammarays doses which killing 50% of malaria parasite populations (IC50). Place and Duration of Study: Department of Biotechnology and Molecular Biology, Department of Biochemistry: General Atomic Energy Commission, Regional Nuclear Studies Center of Kinshasa, P.O. Box. 868 Kinshasa XI, Democratic Republic of the Congo (DRC). National Institute of Biomedical Research (NIBR) at Kinshasa/Gombe (DRC). Laboratory of UMR-MD3, Aix-Marseille University, Faculty of Pharmacy/Marseille, France. The experiments were conducted from Junuary - December 2010; August and December 2011 and during May and September 2012 until July 2014. Methodology: Aerial parts of plants M1 and M2 from Gamma-rays irradiation of P. odontadenius seeds or from immersion of P. odontadenius seeds in sodium azide solutions used as biological material for the in vitro antiplasmodial analysis. The in vitro antiplasmodial activities assays on clinical isolates of P. falciparum and on chloroquine-resistante P. falciparum strain K1 was determined using microscopic method, the isotopic micro-test method and using HRP2-based ELISA assay. Results: Gamma-rays (Cs-137) increased (multiplied) the in vitro antiplasmodial activities from 2.48 up to 7.6 in comparison to control. Thus, the in vitro antiplasmodial activities were improved or exceeded from 147.57% up to 660% than those of control plant. SA had increased (multiplied) the in vitro antiplasmodial activities from 1.24 up to 10.15 comparing to the control plants. The antiplasmodial activities in vitro were exceeded compared to the control plants from 24.43% up to 915%. Conclusion: Treatment of P. odontadenius seeds by Gamma-rays or by SA give plants high in vitro antiplasmodial activities. Values of in vitro antiplasmodial activities varried from 1.24 (147.57%) to 10.15 (915%). 125, 150 and 225 Gy of Gamma-rays (Cs-137) for physical mutagenesis and 10.15 and 17.5 M of SA solutions for the chemical mutagenesis could be used for improving in vitro antiplasmodial activities against P. falciparum (clinical isolates of P. falciparum or chloroquine-resistant P. falciparum strain K1). Thus, plants extracts from treated seeds have justified the usefulness of mutagens in plant breeding particularly in the increasing production of secondary metabolite against malaria parasite.

Phytochemistry and antiplasmodial properties of aqueous and methanol leaf extracts of &lt;i&gt;Jatropa curcas&lt;/i&gt;

This study was aimed at evaluating the phytochemistry and antiplasmodial properties of the aqueous and methanolic leaf extracts of Jatropa curcas against the malaria parasite; Plasmodium falciparum . The need for new therapeutic compounds against malaria parasite is made more urgent by the continued spread of resistance to almost all the available anti malarial drugs. Phytochemical detection followed by antiplasmodial sensitivity test was carried on the two leaf extracts following standard procedures. The pyhtochemicals detected from both extracts were alkaloids, glycosides, tannins, saponins, phenol, steroids and flavonoids. The result of Plasmodium sensitivity tests revealed that both the aqueous and methanolic leaf extracts of Jatropa curcas were highly effective against the malaria parasite. However, the methanolic extract showed greater activity than the aqueous extract. At extract concentration of 10mg/ml, the methanol extract produced the highest parasite clearance rate after 72 hour incubation period, with percentage elimination of 95.4% while the aqueous extract produced 87% elimination at the same concentration and incubation period. These observations showed that Jathropa plant may contain chemicals with promising antimalarial properties which when fully harnessed could constitute a novel therapy for the management of clinical malaria. Keywords : Antiplasmodial, Jatropha Curcas , Extracts, Phytochemistry.