Anthelmintic Activity of Ethanolic and Aqueous Extracts of Khaya grandifoliola Stem Bark against Heligmosomoides polygyrus: In Vitro and In Silico Approaches.

Background: Parasitic infection remains a serious health trade for humans and livestock. The purpose of this study was to present scientific proof of the anthelmintic properties of Khaya grandifoliola, which the native population uses to cure helminthiasis. Method: Fresh Heligmosomoides polygyrus eggs were isolated from faecal samples of experimentally infected mice. The faecal material was cultured, and L1 and L2 larval stages were recovered after 48 and 120 hours, respectively. Using the worm microtracker, the anthelminthic efficacy of the extracts against H. polygyrus was assessed. Two different extracts (aqueous and ethanol extracts) were prepared. For the ovicidal and larvicidal activities, 100 µL of various concentrations of plant extracts, levamisole and 1.5% dimethyl sulfoxide (DMSO), were introduced into a 96-well microplate titer followed by the addition of 100 µL of embryonated eggs (60 eggs) for the ovicidal activity and 100 µL of L 1 or L 2 larvae (50 larvae) for the larvicidal activity. The movement of the worm was monitored for 24 hours in the worm microtracker at 27°C. The Glide module of the Schrodinger Maestro software was used to perform docking studies. Results: For the aqueous extracts, the highest percentage of inhibition of hatching was 42.77 ± 12% at 7.5 mg/mL. The IC50 values for the ethanol (0.36 mg/mL) extract showed that the ethanol extract had a good inhibitory effect on the ability of parasites to hatch from eggs. The inhibition percentage of L1 larvae motility at 7.5 mg/mL was 98.0 ± 1.66% and 83.33 ± 1.66% for ethanol and aqueous extracts, respectively. The negative controls, distilled water and 1.5% DMSO, had no inhibitory impact on larvae. On L1-larvae, the drug of choice levamisole (positive control) had the highest percentage effect (100.0%). Six compounds had the highest docking score and their interactions with the receptor as well. Grandiamide A interacts most with tyrosine, glycine, phenylalanine, asparagine, and serine, and its benzene ring and oxygens inhibit these receptors. Carbonyl and hydroxyl (OH) groups connect grandiamide D to asparagine, isoleucine, and phenylalanine, respectively. By donating hydrogen to the receptor through OH groups, D-glucopyranose-6-phosphate also forms relatively strong hydrogen bonds with its oxygen-bound phosphorus and the receptor. 1-O-deacetylkhayanolide E interacts most with serine and glutamic acid. The carbamic acid benzyl ester of carbamic acid [(1S)-1-phenyl-2-[(4-methylphenyl) sulfinyl] ethyl] interacts most with the receptor with carbonyl groups and with asparagine and serine. With its abundant hydroxide, D-mannitol acts as a hydrogen donor and acceptor and interacts most strongly with amino acids such as glycine, asparagine, aspartic acid, alanine, and glutamic acid. Conclusions: K. grandifoliola extracts possess anthelminthic properties. However, in vivo studies are still necessary to demonstrate the effectiveness of this plant for the treatment of helminthiasis.

In Vitro Anthelmintic Activities of Khaya anthotheca and Faidherbia albida Extracts Used in Chad by Traditional Healers for the Treatment of Helminthiasis and In Silico Study of Phytoconstituents.

Background: Helminthiasis is endemic in Chad and constitutes a public health problem, particularly among school-age children. The aim of this study was to evaluate the anthelmintic activity of extracts of Khaya anthotheca and Faidherbia albida used in Chad by traditional healers for the treatment of helminthiasis. Methods: The anthelmintic activity was assessed against Heligmosomoides polygyrus and Caenorhabditis elegans larvae using the Worm Microtracker. Embryonated eggs, L1, L2, and L3 larvae of H. polygyrus were obtained after 24 h, 48 h, and 7 days of coproculture and L4 larvae of C. elegans culture using standard procedures. One hundred microliters of extracts at various concentrations, with albendazole and distilled water were, put in contact with 100 µL of H. polygyrus suspension (containing 50 parasites at various developmental stages) in a microplate and incubated for 20 h at 25°C in the Worm Microtracker. The same procedure was adopted for C. elegans, but with 180 µL of OP50. 19 µL of C. elegans suspension (containing 50 larvae) was put in contact with 1 µL of extract at various concentrations and incubated in the Worm Microtracker. Docking studies were carried out using the Schrodinger Maestro software's Glide module. The score function in the software was used to rank and group distinct possible adduct structures generated by molecular docking. Results: The aqueous and ethanolic extracts of F. albida at a concentration of 2.5 mg/mL showed the same activity as albendazole (100 ± 0.00) on hatching. The IC50s of the aqueous extracts of the two plants (IC50: 0.6212 mg/mL and 0.71 mg/mL, respectively) were comparable on egg hatching of H. polygyrus with no significant difference (p ≥ 0.05) with respect to the ethanol extracts (IC50: 0.70 mg/mL and 0.81 mg/mL, respectively). There was no significant difference between the percentage inhibition of extracts and albendazole on the L1 larvae of H. polygyrus (p ≥ 0.05). The aqueous extracts acted more effectively than the ethanol extracts on the L1 larvae of H. polygyrus with an IC50 of 0.5588 and ∼9.858e - 005 mg/ml, respectively, for K. anthotheca and F. albida. The aqueous extracts of K. anthotheca and F. albida on L3 larvae of H. polygyrus had inhibitory percentages of 92.6 ± 0.62 and 91.37 ± 0.8 at 2.5 mg/mL which were lower than albendazole (100 ± 0.00). The aqueous extracts of K. anthotheca and F. albida on C. elegance showed IC50 of 0.2775 µg/mL and 0.5115 µg/mL, respectively, and were more effective than the ethanol extracts. Examining K. anthotheca and F. albida through the interaction with the protein receptor and its results also confirmed our assumption that the compound used has hydroxyl and carbonyl groups as well as aromatic rings and is exposed to phenolic and flavonoid groups in a more specific way, and it shows a better inhibitory effect. Conclusions: This study scientifically validates the use of extracts of the two plants in the traditional treatment of helminthiasis. However, it will be necessary to evaluate the in vivo anthelmintic activity and toxicity. Examining the ADME properties of these compounds also supports the potential of these ligands to be transformed into pharmaceutical forms.

In Vitro Antiplasmodial, Cytotoxicity, and Antioxidant Activities of Lophira lanceolata (Ochnaceae): A Cameroonian Plant Commonly Used to Treat Malaria.

Background: Malaria is the leading cause of morbidity and mortality in African countries. We aimed this study at evaluating the in vitro antiplasmodial, antioxidant, and cytotoxicity activity of Lophira lanceolata extracts. Method: The aqueous and ethanol extracts were obtained by maceration. It tested in vitro the extracts against Plasmodium falciparum 3D7 and multiresistance Dd2. Macrophage cell lines (RAW 264.7 cells) and red blood cells were used for cytotoxicity tests. The antioxidant activity was assessed by 1,1-diphenyl-2-picrylhydrazine (DPPH), hydrogen peroxide (H2O2), nitric oxide (NO) reduction, and ferric reducing antioxidant power (FRAP) scavenging. Results: The in vitro antiplasmodial results showed that the ethanol extract was the most active, with IC50 of 24.51 ± 4.77 µg/mL and 31.86 ± 3.10 µg/mL, respectively, on the resistant Dd2 and sensitive 3D7 strains unlike the aqueous which indicated moderate activity with an IC50 of 51.36 ± 4.86 µg/mL and 56.36 ± 4.27 µg/mL, respectively, on the resistant Dd2 and sensitive (3D7) strains. However, the ethanol extract had the highest activity, with an IC50 of 8.153 g/mL, 1915 g/mL, 30.81 g/mL, and 54.66 g/mL, respectively, for DPPH, H2O2, NO, and FRAP, while the aqueous extract had an IC50 of 6.724, 2387681, 185.7, and 152.0 g/mL, respectively, for DPPH, H2O2, NO, and FRAP. The cytotoxicity test reveals that both extracts do not promote red blood cell haemolysis. They presented weak activity against RAW 264.7 cells and red blood cells. Conclusion: According to these findings, the aqueous and ethanol extracts have antiplasmodial and antioxidant activity but with no cytotoxic effects on red blood cells or RAW cells. However, it will be important to investigate the in vivo antiplasmodial and antioxidant activity of these extracts.

Antihelminthic Activity of Lophira Lanceolata on Heligmosomoides polygyrus Using an Automated High-Throughput Method.

Background: There are about 13 parasitic infections that are responsible for significant morbidity and mortality but have not received the attention they deserve; thus, they are now known as "neglected tropical diseases" (NTDs). This study was aimed at evaluating the antihelminthic activities of Lophira lanceolata using an automated high-throughput method. Methods: The antihelminthic activity effect of the extracts against H. polygyrus was determined using an automated high-throughput method. For the egg-hatching test, 100 µL of embryonated egg suspension (60 eggs) was added to 100 µL of various concentrations of extracts, levamisole, and 1.5% DMSO in a 96-well round-bottom microtitre plate. After mixing, the 96-well microplate was placed in WMicroTracker and incubated for 24 h at 25°C; the movements were recorded every 30 minutes. The same procedure was used for the larval motility assays, where 100 µL of L1 or L2 larvae (50 larvae) were put in contact with 100 µL of various concentrations of extracts. Results: The ovicidal activity (hatching) had an IC50 of 1.4 mg/mL for the ethanol extract. The aqueous and ethanol extracts of L. lanceolata showed larvicidal activity on the L1 larvae with IC50 of 1.85 mg/mL and 2.4 mg/mL, respectively, as well as on the L2 larvae with IC50 values of 1.08 mg/mL and 1.02 mg/mL for the aqueous and ethanol extracts, respectively. These results showed that the aqueous extract exhibited a stronger inhibitory power on the hatching rate of parasites than ethanol extracts, while the contrary effect was observed for the larval motility assays. Conclusion: This study provides scientific data on the use of L. lanceolata by the local population for the treatment of helminthiases. However, in vivo and toxicity tests are necessary to assess its activity and safety.

Antiplasmodial, Antioxidant and Cytotoxicity Activity of Ethanol and Aqueous Extracts of Khaya grandifoliola Stem Bark.

Background: Malaria is a serious public health problem, especially in sub-Saharan Africa. The aim of this study was to scientifically provide baseline information on the use of Khaya grandifoliola stem bark as an antimalaria drug by traditional healers. Method: The stem barks of K.grandifoliola were harvested and dried to obtain powder, and fifty grams of the powder were soaked in ethanol and hot distilled water respectively, for the preparation of ethanol and aqueous extracts, then dried in an oven at 40°C for the ethanol extract and 50°C for the aqueous extract. Plasmodium falciparum strains 3D7 sensitive and Dd2 resistant to chloroquine, were used to evaluate in vitro antiplasmodial activity using SYBR Green. The ability of the extracts to prevent oxidative stress was assessed by trapping 2, 2'-diphenyl-1-picrylhydrazyl (DPPH); nitric oxide, hydrogen peroxide and ferric reducing power. The cytotoxicity test of the extracts was carried out on RAW 264.7 cell lines and on erythrocytes. The data obtained were entered in the Excel software, then in Graph pad where the IC50 was calculated and the curves plotted. Results: The fifty percent inhibition (IC50) of the antiplasmodial activity of the chloroquine-resistant strain PfDd2 were 54.27 ± 2.41 µg/mL and 31.19 ± 4.06 µg/mL respectively, for the aqueous and ethanol extracts. As for the Chloroquino-sensitive Pf3D7, IC50 of 53.06 µg/mL was obtained for the aqueous extract and 28.03 ± 1.90 µg/mL for ethanol. The DPPH radical scavenging activity presented IC50 of 104 µg/mL for the aqueous and 2.617 µg/mL for the ethanol extract; for the Nitric oxide (NO) presented an IC50 of 301 ± 21 µg/mL for the aqueous extract 140.7 ± 21 µg/mL for the ethanol; for hydrogen peroxide the ethanol and aqueous presented IC50 of 845.1 ± 21 µg/mL and 509.4 ± 21 µg/mL respectively. The cytotoxicity on RAW 264.7 cells presented High CC50 in particular >1000 µg/mL and 467.4 µg/mL respectively for the aqueous and ethanol extract. Conclusion: Extracts of Khaya grandifoliola exhibited antiplasmodial activity. The ability to inhibit oxidative stress as well as lower cell toxicity on RAW 264.7 and erythrocytes, is a good indicator. However, in vivo tests remain important in order to confirm the use of this plant for the treatment of malaria.

Risk Factors Associated with Prevalence of Candida albicans, Gardnerella vaginalis, and Trichomonas vaginalis among Women at the District Hospital of Dschang, West Region, Cameroon.

BACKGROUND: Vaginal or genitourinary infections are a major cause of morbidity, sterility, and increase in the vulnerability to cancers and HIV/AIDS infection. The aim of this study was to determine the prevalence of vaginal infections of C. albicans, G. vaginalis, and T. vaginalis among women in the locality of Dschang, West Region of Cameroon. METHOD: A prospective study was carried out in the District Hospital of Dschang. After obtaining informed consent, one thousand and one (1001) samples of vaginal swabs were collected. Biological diagnosis was carried out on fresh samples, Gram stained, and then cultivated in Sabouraud agar in a Petri dish, in order to isolate and identify the various infectious agents. RESULTS: Five hundred and twenty-five (525) women were diagnosed positive, hosting at least one of these microorganisms, making an overall prevalence of 52.44%. Two hundred and fifty-six (256) women (25.57%) were infected with C. albicans, and 171 (17.08%) with G. vaginalis. Ninety-five (9.49%) were infected with both C. albicans and G. vaginalis, 2 (0.20%) with C. albicans and T. vaginalis, and 1 (0.1%) with G. vaginalis and T. vaginalis. CONCLUSION: Drastic measures should be taken in order to improve life styles to regress the frequency of these infections. Results obtained in this study, will help to educate and shed more light on the prevalence of vaginal infections in the West Region of Cameroon.