Ex vivo Study of Antimalarial Activity of Canarium Odontophyllum Leaf Extracts Against Plasmodium Berghei NK65

2021 ◽  
Vol 1 ◽  
Author(s):  
Shafariatul Akmar-Ishak ◽  
Fifi Fariza-Azmi ◽  
Adibah Syahnaz-Zahari ◽  
Dayang Fredalina-Basri
Keyword(s):  
Plasmodium Berghei ◽  
Antimalarial Activity ◽  
Ex Vivo ◽  
Sybr Green ◽  
Fluorescence Assay ◽  
Sybr Green I ◽  
Leaf Extracts ◽  
Significant Difference ◽  
The One ◽  
Canarium Odontophyllum

Background: Malaria is a parasite that is transmitted to human through the bite of a female Anopheles mosquito. Every year human was exposed to the threat of malaria infection. This disease becomes more fatal as these parasites show resistance towards the drug available. Thus, searches for new antimalarial drug are crucial. This study was carried out to evaluate the antimalarial activity in Canarium odontophyllum leaf extracts (methanol, acetone and aqueous) against erythrocytes infected with Plasmodium berghei NK65 using Plasmodium Lactate Dehydrogenase (pLDH) Assay and SYBR green I fluorescence Assay. Method: Three types of solvents were used to extracts Canarium odontophyllum leaf according to increasing polarity index; acetone, methanol and aqueous. These extracts were made into eight-fold serial dilution; concentrations ranging from 0.00001μg / ml as the lowest concentration until 100 μg / ml as the highest concentration and further tested on Plasmodium berghei NK65 infected erythrocytes via ex-vivo. The IC50 (inhibition concentration) 50 readings were taken at the point of 5% parasitemia level and in the synchronization process. Both PLDH assay and SYBR green I fluorescence assay were being carried out simultaneously. Result: The One-way ANOVA showed that there is no significant difference between extracts at 5% parasitemia level, even so methanol was further tested on synchronization process as it showed the lowest reading of IC50 among the three extracts for PLDH assay and SYBR green I fluorescence assay respectively, (IC50 0.00045μg / ml, 0.002 μg / ml). For synchronization stages, the One-way ANOVA result showed there is no significant difference between stages of morphology. However, methanol extracts showed the most potent on schizont, (1.16x10-5 μg / ml ) and young trophozoite, (0.00195 μg / ml ) stages for each method respectively. Conclusions: All three extracts of Canarium odontophyllum leaf were effective on Plasmodium berghei NK65, however methanol showed most promising results and further research on the fractions were required for proper drug development. The Ministry of Higher Learning funded this project, Government of Malaysia, under the Fundamental Research Grant Scheme Code No. FRGS/2/2014/SG05/UKM/02/3.

scholarly journals Assessment of SYBR Green I Dye-Based Fluorescence Assay for Screening Antimalarial Activity of Cationic Peptides and DNA Intercalating Agents

2015 ◽  
Vol 59 (5) ◽  
pp. 2886-2889 ◽  
Author(s):  
Rakesh Bhatia ◽  
Ankur Gautam ◽  
Shailendra K. Gautam ◽  
Divya Mehta ◽  
Vinod Kumar ◽  
...  
Keyword(s):  
Antimalarial Activity ◽  
Cell Penetrating Peptides ◽  
Sybr Green ◽  
Fluorescence Assay ◽  
Sybr Green I ◽  
Cationic Peptides ◽  
Binding Agents ◽  
Dna Binding Agents ◽  
Incorporation Assay ◽  
Intercalating Agents

ABSTRACTThe SYBR green I (SG) dye-based fluorescence assay for screening antimalarial compounds is based on direct quantitation of parasite DNA. We show that DNA-interacting cationic cell-penetrating peptides (CPPs) and intercalating agents compete with SG dye to bind to DNA. Therefore, readouts of this assay, unlike those of the [3H]hypoxanthine incorporation assay, for the antimalarial activity of the above DNA binding agents may be erroneous. In the case of CPPs, false readouts can be improved by the removal of excess peptides.

Label-free fluorescence assay coupled exonuclease reaction and SYBR Green I for the detection of T4 polynucleotide kinase activity

2020 ◽  
Vol 12 (6) ◽  
pp. 807-812
Author(s):  
Xu Wu ◽  
Shuyi He ◽  
Julia Xiaojun Zhao
Keyword(s):  
Kinase Activity ◽  
Sybr Green ◽  
Fluorescence Assay ◽  
Sybr Green I ◽  
Label Free ◽  
Cleavage Reaction ◽  
Polynucleotide Kinase ◽  
T4 Polynucleotide Kinase

A sensitive label-free fluorescence assay for monitoring T4 polynucleotide kinase (T4 PNK) activity and inhibition was developed based on a coupled λ exonuclease cleavage reaction and SYBR Green I.

scholarly journals Study of the Antimalarial Activity of the Leaf Extracts and Fractions of Persea americana and Dacryodes edulis and Their HPLC Analysis

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Philip F. Uzor ◽  
Chukwuebuka K. Onyishi ◽  
Adaeze P. Omaliko ◽  
Somtochukwu A. Nworgu ◽  
Onyemaechi H. Ugwu ◽  
...  
Keyword(s):  
Plasmodium Berghei ◽  
Hplc Analysis ◽  
Antimalarial Activity ◽  
Persea Americana ◽  
Leaf Extracts ◽  
Lead Compounds ◽  
Phytochemical Composition ◽  
Potential Sources ◽  
Water Fractions ◽  
Dacryodes Edulis

In the present study, the antimalarial activity of the extracts and fractions of the leaves of Persea americana and Dacryodes edulis as well as their phytochemical compositions were examined. Each of the extracts of the plants was successively fractionated to obtain hexane, ethyl acetate, methanol, and water fractions. The extracts and fractions were tested against Plasmodium berghei in both curative and suppressive antimalarial mouse models. Their major phytochemical composition was studied by the standard chemical tests and HPLC analysis. The extracts and fractions of P. americana and D. edulis demonstrated significant ( p < 0.05 ) maximal plasmodial inhibition as 52.16 ± 2.77% and 57.10 ± 1.98%, respectively, and chemosuppression of parasitemia as 64.01 ± 0.08% and 71.99 ± 0.06%, respectively. The major secondary metabolites identified in the plants include alkaloids, flavonoids, and saponins. It was concluded that P. americana and D. edulis possess promising antimalarial activity and they are potential sources of new lead compounds against malaria.

scholarly journals Antimalarial Activity of Cordia africana (Lam.) (Boraginaceae) Leaf Extracts and Solvent Fractions in Plasmodium berghei-Infected Mice

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Dawit Zewdu Wondafrash ◽  
Dayananda Bhoumik ◽  
Birhanetensay Masresha Altaye ◽  
Helen Bitew Tareke ◽  
Brhane Teklebrhan Assefa
Keyword(s):  
Acute Toxicity ◽  
Plasmodium Berghei ◽  
Antimalarial Activity ◽  
Lethal Dose ◽  
Public Health Problem ◽  
Chloroform Fraction ◽  
Leaf Extracts ◽  
Suppression Test ◽  
Cordia Africana

Background. Malaria remains a major worldwide public health problem leading to death of millions of people. Spread and emergence of antimalarial drug resistance are the major challenge in malaria control. Medicinal plants are the key source of new effective antimalarial agents. Cordia africana (Lam.) is widely used for traditional management of malaria by local people in different parts of Ethiopia. The present study aimed to evaluate in vivo antimalarial effects of leaf extracts and solvent fractions of Cordia africana on Plasmodium berghei-infected mice. Methods. The leaf extracts were prepared and tested for oral acute toxicity according to the OECD guideline. In vivo antimalarial effects of various doses of C. africana extracts and solvent fractions were determined using the four-day suppression test (both crude and fractions), as well as curative and chemoprophylactic tests (crude extracts). Results. The acute toxicity test of the plant extract revealed that the medium lethal dose is higher than 2000 mg/kg. The crude extract of the plant exhibited significant parasitemia suppression in the four-day suppression (51.19%), curative (57.14%), and prophylactic (46.48%) tests at 600 mg/kg. The n-butanol fraction exhibited the highest chemosuppression (55.62%) at 400 mg/kg, followed by the chloroform fraction (45.04%) at the same dose. Conclusion. Our findings indicated that both the crude leaf extracts and fractions of C. africana possess antimalarial effects, supporting the traditional claim of the plant.

Flow cytometric enumeration of Plasmodium berghei-infected red blood cells stained with SYBR Green I

Acta Tropica ◽  
2012 ◽  
Vol 122 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Voravuth Somsak ◽  
Somdet Srichairatanakool ◽  
Yongyuth Yuthavong ◽  
Sumalee Kamchonwongpaisan ◽  
Chairat Uthaipibull
Keyword(s):  
Red Blood Cells ◽  
Plasmodium Berghei ◽  
Blood Cells ◽  
Sybr Green ◽  
Sybr Green I ◽  
Flow Cytometric

scholarly journals Antimalarial Activity of Meriandra dianthera Leaf Extracts in Plasmodium berghei-Infected Mice

2020 ◽  
Vol 2020 ◽  
pp. 1-8 ◽  
Author(s):  
Kalay Hagazy ◽  
Gereziher G. Sibhat ◽  
Aman Karim ◽  
Gebretsadkan H. Tekulu ◽  
Gomathi Periasamy ◽  
...  
Keyword(s):  
Plasmodium Berghei ◽  
Leaf Extract ◽  
Antimalarial Activity ◽  
Negative Control ◽  
Mice Model ◽  
Oral Toxicity ◽  
Leaf Extracts ◽  
Traditional Use ◽  
Crude Leaf Extract

Objective. To evaluate the antimalarial effect of aqueous methanolic extract and solvent fractions of Meriandra dianthera leaves against Plasmodium berghei in mice model. Method. M. dianthera leaves were extracted with 80% methanol and dried. The dried crude extract was then defatted and further fractionated with chloroform, ethyl acetate, and butanol. Acute oral toxicity test was performed as per the Organization for Economic Cooperation and Development guideline 425. Peter’s 4-day suppressive test was used to determine the in vivo antimalarial activity of the extract and fractions. Result. The crude leaf extract of Meriandra dianthera showed parasite inhibition of 42.28% and 45.52% at doses of 400 and 600 mg/kg, respectively, as compared to the negative control. Moreover, the mice which received chloroform and aqueous fractions at the dose of 400 mg/kg/day showed significant (P<0.001) chemosuppression compared to the negative control. Both the extract and fractions were able to prevent P. berghei induced body weight loss and body temperature reduction and also increased the survival time of the mice as compared to the negative control. The aqueous methanolic leaf extract of M. dianthera showed no gross signs of toxicity or mortality in mice until a single oral dose of 2000 mg/kg. Conclusion. The extracts of M. dianthera leaves showed promising antimalarial activity, with no sign of toxicity and therefore may support its traditional use for the treatment of malaria.

Microplate fluorescence assay for the quantification of double stranded DNA using SYBR Green I dye

2006 ◽  
Vol 28 (19) ◽  
pp. 1587-1594 ◽  
Author(s):  
Johanna Leggate ◽  
Ray Allain ◽  
Leah Isaac ◽  
Burton W. Blais
Keyword(s):  
Sybr Green ◽  
Fluorescence Assay ◽  
Sybr Green I ◽  
Double Stranded Dna ◽  
Microplate Fluorescence Assay

scholarly journals Oral acute toxicity and antimalarial potentials of aqueous and methanolic extracts of roots, leaves and stem of Dictyandra arborescens (Welw.) on Plasmodium berghei infected mice

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Uchechi E. Enenebeaku ◽  
Nnamdi C. Ukwandu ◽  
Ifeyinwa C. Mgbemena ◽  
Harriet C. Nwigwe ◽  
Conrad K. Enenebeaku ◽  
...  
Keyword(s):  
Acute Toxicity ◽  
Plasmodium Berghei ◽  
Antimalarial Activity ◽  
Scientific Evidence ◽  
Negative Control ◽  
Methanol Extracts ◽  
Methanolic Extracts ◽  
Ms Analysis ◽  
Significant Difference ◽  
Oral Acute Toxicity

Abstract Background Malaria is one of the tropical diseases of universal concern particularly with continuous appearance of resistant strains of P.falciparum. This calls for continous screening of traditional plants such that new and effective antimalarial agents will be developed. This study therefore explored the oral acute toxicity and antimalarial potentials of aqueous and methanolic extracts of roots, leaves and stem of Dictyandra arborescens on Plasmodium berghei infected mice. Results No mortality was recorded in any of the experimental animal groups even at the highest tested dose (5000 mg/kg b.wt) of the extract after monitoring them for 4hrs and subsequently for 7 days. Out of the six extracts, methanolic extracts of the roots and leaves exhibited more antimalarial activity than others. A significant difference (P < 0.05) was statistically observed in the parasite count of groups that received methanol extracts of roots and leaves of D. arborescens. This observation was made when these two extracts were compared with other groups as well as the negative control. However, activity of the standard antimalarial drug (artesunate) was higher (p˂0.05) than those of the extracts. Phytochemicals such as tannins, alkaloids, saponins, terpenoids, flavonoids etc. were present in the extracts in varying quantities. GC–MS analysis of methanol extract of the root of this plant showed different chemical compounds. Conclusion Administration of aqueous and methanol extracts of roots, leaves and stem of D. arborescens in mice is not harmful at any dose less than or equal to 5000 mg/kg. Methanol extracts exhibited more antimalarial activity than aqueous extracts suggesting that antimalarial activity of the plant parts could be affected by the solvent used for extraction and antimalarial activity may be more in a particular part of a plant. The presence of different bioactive compounds identified in phytochemical and GC–MS analysis could be the fundamental scientific evidence for the antimalarial activity exhibited by this plant especially in the root.

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