Acute and subacute toxicity assessment of liquid CO2 extract of Phaleria macrocarpa fruits flesh in mice model

Objectives Phaleria macrocarpa (Scheff.) Boerl., an evergreen tree is found in Indonesia, Malaysia and Oceania, explicitly Northern Territory of Australia and Papua New Guinea. The fruits are traditionally used to treat various ailments including diabetes, psoriasis, looseness of the bowels, skin inflammation, malignancy, kidney, liver and heart sicknesses. However, to date, no comprehensive study investigating its toxicity profile utilizing liquid CO2 extract (LCE) has been reported in ICR mice. Hence, this research was performed to investigate the both acute and sub-acute toxicities of the liquid carbon dioxide extract from ripe fruit flesh of the P. macrocarpa in mice. Methods Acute toxicity was assessed at a single dose of 3000 mg/kg bw for 2 weeks according to the up and down method of OECD 425 guideline, while the sub-acute test was assessed by liquid carbon dioxide extract at doses of 250, 500, 1000, and 2000 mg/kg bw for 28 days. The animals’ general behaviour, food intake, body weight, organ coefficients, biochemical and histological morphology were observed and analysed. Results After acute exposure to LCE, there was no evidence of any severe or fatal effects in the mice. An acute toxicity result exhibited that LD50 of LCE was > 3000 mg/kg bw. The findings of subacute toxicity evaluation revealed that LCE at doses of 250 and 500 mg/kg bw had no significant detrimental impact. Moreover, LCE at 1000 and 2000 mg/kg bw demonstrated toxicity to the heart, liver, kidney and lung in mice identified by histological and biochemical investigation. Conclusions The results confirmed that the LCE dosage at 500 mg/kg bw is considered a safe dose that can provide a better therapeutic effect without eliciting any adverse side effects

The therapeutic potential of skin mucus from Asian swamp eel (Monopterus albus): In vivo evaluation and histological evidence

Objectives The Asian swamp eel (Monopterus albus), is commonly distributed in Asian countries. However, its therapeutic potential has not been thoroughly investigated yet. The current study aimed to evaluate the in-vivo therapeutic properties of the skin mucus of this fish. Methods The eel mucus was collected fleshly and topical gel with carbopol 934 was formulated to study the antibacterial activity on the infected skin of the rats. Sprague Dawley rats were used in the study and divided into 4 groups negative, positive, normal control, and treated groups. Results Intracutaneous injections of pathogenic bacteria (Streptococcus pyogenes, Staphylococcus aureus) and fungi (Microsporum gypseum, Candida albicans) were injected into the rats. The development of tinea capitis, impetigo, and cutaneous candidiasis in the animal model was confirmed based on clinical and histopathological observations. To treat the infected rats, a formulated gel of eel skin mucus was applied on the infected rat’s skins topically. The histological analysis confirms a complete recovery in the skin tissues similar to commercial antifungal and antibacterial agents used in the positive control groups. Conclusion The present novel eel skin mucus is an efficient therapeutic candidate in treating skin infections associated with pathogenic microbes

Design, synthesis, in vitro antiproliferative effect and in situ molecular docking studies of a series of new benzoquinoline derivatives

Quinoline derivatives have been reported to possess multi-therapeutic potential owing to the manifestations of different pharmacological effects. The current research work describes about the design and synthesis of a series of novel benzoquinoline analogues with an objective to evaluate their antiproliferative structure–activity relationship against colon, breast and hepatocellular cancers. Upon synthesis, all derivatives’ chemical structures were elucidated through FTIR, 1 HNMR and 13CNMR spectroscopic analysis. All derivatives were investigated for their in vitro anti-proliferative property against three different cancer cell lines (viz., colon carcinoma HT29, Caucasian breast adenocarcinoma MCF7, hepatocellular carcinoma HepG2) and a normal non-transformed human foreskin fibroblast Hs27 cell line. All derivatives demonstrated varied degrees of strong anticancer effect against all of the cell lines with the 2-Amino-4-(4-nitrophenyl)-5,6-dihydrobenzo[h]quinoline-3-carbonitrile (CNMP, 2) exhibited the most potent antiproliferative effect viz. LC50 21.23 lM for breast, 8.24 lM for colon, and 26.15 lM for the hepatocellular, respectively. Molecular docking studies against all the the target crystal structures of cancer proteins (1HK7, 3EQM, 3IG7 and 4FM9) revealed significant binding affinities via hydrophobic and H-bonding interactions with all the compounds in conformity with the wet lab results. CNMP showed the highest binding energy of �7.55 in the HT29, �6.9 (both in MCF7 HepG2) kcal/mol. Based on the results obtained from wet lab and dry lab experiments, it can be proposed that CNMP might prove to be a potential lead structure for the design and synthesis of more potent anticancer candidates

Anti-obesity and antihyperlipidemic effects of Phaleria macrocarpa fruit liquid CO2 extract: In vitro, in silico and in vivo approaches

Objective: Phaleria macrocarpa fruit has been reported to be effectively used in Malaysia and neighboring countries to prevent obesity. Despite its anti-obesity potential, no research has ever predicted the compound-lipase, compound-HMG-CoA reductase interactions and in vivo investigations to further confirm its anti-obesity properties. Thus, the purpose of this study was to assess the anti-obesity and antihyperlipidemic effects of P. macrocarpa by in vitro, computational (in silico), and in vivo assays. Method: Initially, fruit was extracted through liquid CO2 and heating under reflux extraction methods to obtain liquid CO2 extracts (LCE-1 and LCE-2) and heating under reflux extract (HRE), respectively. Subsequently,allthreeextractswereassessedforantioxidantpotentialsthroughinvitrobioassaysand FTIRanalysistoidentifydifferenttypesoffunctionalgroupspresentintheexistingbioactivecompounds. Successively,themostactivefraction(LCE-2)wassubjectedtoGCMSanalysisfortheidentificationof lipidloweringandlipaseinhibitorsthroughmoleculardockingapproach.Finally,anti-obesityandlipid loweringeffectswerefurtherconfirmedthroughaninvivoassayusingmice. Result:LCE-2exhibitedhigherDPPH(IC50=0.172mg/mL)andFRAP(78.98AAEmg/g)antioxidantactivitiesandshowedmorepotentpeaksinFTIRchromatogramsthanLCE-1andHRE.Amongtheidentified tentativebioactivecompoundsviz.methylpalmitate,palmiticacid,ethylpalmitate,methyloleate,oleic acid,cis-vaccenicacid,3-deoxyestradiolandphenol,2,20-methylenebis[6-(1,1-dimethylethyl)-4-methylwerefoundasananti-obesityandlipidloweringcompoundswhoseprotein–ligandinteractionwasconfirmedbybindingaffinity,aminoacidresiduesandbondinginteractions.Similarly,anti-obesityandlipid loweringfindingswerealsofoundinamicemodelafter6weekstreatmentatadoseof250mg/kgb.w. Conclusion:Basedontheaforementionedinvitro,insilicoandinvivofindings,itisconcludedthatthe LCE-2possesseslipaseandHMG-CoAreductaseinhibitorsthatcanassisttodevelopthisplant’sextract asanalternativesafelipidloweringherbalmedicineinfuture

Neurotoxicity of aluminium chloride and okadaic acid in zebrafish: Insights into Alzheimer's disease models through anxiety and locomotion testing, and acute toxicity assessment with Litsea garciae bark's methanolic extract

Alzheimer's disease (AD) is a complicated neurodegenerative disorder that presents significant challenges for the development of effective therapeutic interventions. Understanding disease mechanisms and exploring potential treatments require the use of animal models that accurately replicate the pathology of AD. In this study, we investigated the potential of two neurotoxin inducers, aluminium chloride (AlCl3) and okadaic acid (OKA), to validate the zebrafish as a model organism for AD. AD can impact locomotor activity and induce anxiety-like behaviors. To assess these behaviors, a 6-minute novel tank test was conducted. Zebrafish were administered with low, medium, or high doses of neurotoxic agent (AlCl3 or OKA) intraperitoneally twice weekly for 21 days. Behavioral activities were recorded at three time points: day 7 (short duration), day 14 (moderate duration), and day 21 (extended duration). The behavioral task required the evaluation of four endpoints. Methanolic extract of Litsea garciae bark was selected as a potential plant for the treatment of AD in this study, based on its previously demonstrated antioxidant effect. However, the acute toxicity of this plant has not been previously assessed. Therefore, this research was aimed to investigate the acute toxicity of the L. garciae bark’s methanolic extract in adult zebrafish. The extract was immersed in a static system following OECD Test Guideline No. 203, and the acute toxicity test involved monitoring the adult zebrafish for 96 h for any deaths or apparent abnormalities. Regarding the behavioural task, the groups induced with 100 nM of OKA demonstrated significant differences in all measured parameters compared to the control group at the 21-day time point. In contrast, none of the parameters were significantly different between the AlCl3-induced groups and the control group at any of the three time points (7, 14, or 21 days). Regarding acute toxicity, neither the test group (100 mg/L) nor the control group recorded any deaths or abnormalities. Therefore, no LC50 value could be determined. These findings confirm the acceptance of OKA as an inducer in the zebrafish model of AD and highlight the significance of the safe and non-toxic nature of L. garciae bark's methanolic extract for future ethnopharmacological investigations

Neurotoxicity of aluminium chloride and okadaic acid in zebrafish: insights into alzheimer's disease models through anxiety and locomotion testing, and acute toxicity assessment with Litsea garciae bark's methanolic extract

Alzheimer's disease (AD) is a complicated neurodegenerative disorder that presents significant challenges for the development of effective therapeutic interventions. Understanding disease mechanisms and exploring potential treatments require the use of animal models that accurately replicate the pathology of AD. In this study, we investigated the potential of two neurotoxin inducers, aluminium chloride (AlCl3) and okadaic acid (OKA), to validate the zebrafish as a model organism for AD. AD can impact locomotor activity and induce anxiety-like behaviors. To assess these behaviors, a 6-minute novel tank test was conducted. Zebrafish were administered with low, medium, or high doses of neurotoxic agent (AlCl3 or OKA) intraperitoneally twice weekly for 21 days. Behavioral activities were recorded at three time points: day 7 (short duration), day 14 (moderate duration), and day 21 (extended duration). The behavioral task required the evaluation of four endpoints. Methanolic extract of Litsea garciae bark was selected as a potential plant for the treatment of AD in this study, based on its previously demonstrated antioxidant effect. However, the acute toxicity of this plant has not been previously assessed. Therefore, this research was aimed to investigate the acute toxicity of the L. garciae bark’s methanolic extract in adult zebrafish. The extract was immersed in a static system following OECD Test Guideline No. 203, and the acute toxicity test involved monitoring the adult zebrafish for 96 h for any deaths or apparent abnormalities. Regarding the behavioural task, the groups induced with 100 nM of OKA demonstrated significant differences in all measured parameters compared to the control group at the 21-day time point. In contrast, none of the parameters were significantly different between the AlCl3-induced groups and the control group at any of the three time points (7, 14, or 21 days). Regarding acute toxicity, neither the test group (100 mg/L) nor the control group recorded any deaths or abnormalities. Therefore, no LC50 value could be determined. These findings confirm the acceptance of OKA as an inducer in the zebrafish model of AD and highlight the significance of the safe and non-toxic nature of L. garciae bark's methanolic extract for future ethnopharmacological investigations

Potential anticancer agents identification of Hystrix brachyura bezoar through gas chromatography-mass spectrometry-based metabolomics and protein-ligand interaction with molecular docking analyses

Background: Bezoar (PB) is a rare, solidified form of undigested food commonly found in the gastrointestinal tract of porcupine (Hystrix brachyura). It is believed to be traditionally used to treat various diseases including different kinds of cancers in Malaysia. However, its active principles have not been found out yet. The purpose of this study was to investigate the anticancer property of PB extract as well as to identify the metabolites responsible for its anticancer effect through a widely acclaimed metabolomics approach. Methods: Initially, 25 PB extracts using various solvent ratios of methanol–water (100, 75, 50, 25, 0% v/v) were prepared in regard to metabolomics approach and subsequently the cytotoxicity of each extract was determined against (melanoma) A375 cell line. The metabolites profiling of the most potent extract was conducted using gas chromatography mass spectrometry (GC–MS) and in silico investigation was performed on Bcl-2 and cyclin/CDK1 complex protein. Results: The correlation of the bioactivity with GC–MS data produced an orthogonal partial least square (OPLS) model which pinpointed four putative active compounds namely (1) cholest-7-en-3-beta-ol,4,4- dimethyl-,acetate; (2) 4-androsten-4-ol-3,17-dione; (3) isolongifolol and (4) gallic acid. The in silico data suggested the binding score and binding mode of active metabolites with the amino acid residues of protein via hydrophobic interactions. Conclusion: This study is the first to report the identified anticancer compounds from PB extract and evaluate them using molecular docking. This further confirms and justifies its traditional usage as an alternative medicine for the treatment of cancer in Malaysia

Potential anticancer agents identification of Hystrix Brachyura Bezoar through gas chromatography-mass spectrometry-based metabolomics and protein-ligand interaction with molecular docking analyses

Background: Bezoar (PB) is a rare, solidified form of undigested food commonly found in the gastrointestinal tract of porcupine (Hystrix brachyura). It is believed to be traditionally used to treat various diseases including different kinds of cancers in Malaysia. However, its active principles have not been found out yet. The purpose of this study was to investigate the anticancer property of PB extract as well as to identify the metabolites responsible for its anticancer effect through a widely acclaimed metabolomics approach. Methods: Initially, 25 PB extracts using various solvent ratios of methanol–water (100, 75, 50, 25, 0% v/v) were prepared in regard to metabolomics approach and subsequently the cytotoxicity of each extract was determined against (melanoma) A375 cell line. The metabolites profiling of the most potent extract was conducted using gas chromatography mass spectrometry (GC-MS) and in silico investigation was performed on Bcl-2 and cyclin/CDK1 complex protein. Results: The correlation of the bioactivity with GC-MS data produced an orthogonal partial least square (OPLS) model which pinpointed four putative active compounds namely (1) cholest-7-en-3-beta-ol,4,4-dimethyl-,acetate; (2) 4-androsten-4-ol-3,17-dione; (3) isolongifolol and (4) gallic acid. The in silico data suggested the binding score and binding mode of active metabolites with the amino acid residues of protein via hydrophobic interactions. Conclusion: This study is the first to report the identified anticancer compounds from PB extract and evaluate them using molecular docking. This further confirms and justifies its traditional usage as an alternative medicine for the treatment of cancer in Malaysia

Exploring the Anti-Corrosion, Photocatalytic, and Adsorptive Functionalities of Biogenically Synthesized Zinc Oxide Nanoparticles

This study reported the synthesis of ZnO nanoparticles (ZnO NPs) using Cucurbita pepo L. seed extract and explored their multifunctional properties such as anti-corrosion, photocatalytic, and adsorption capabilities. The synthesized ZnO NPs were characterized by Fourier-transform infrared spectroscopy (FTIR) to identify their functional groups, thermogravimetric analysis (TGA) to assess their thermal stability, transmission electron microscopy (TEM), and scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDX) to determine their size, morphology, and elemental composition. The characterization of biofabricated ZnO NPs revealed an average particle size of 32.88 nm; however, SEM displayed a tendency for the particles to agglomerate. Furthermore, the X-ray diffraction (XRD) and EDX analysis confirmed the NPs as ZnO, matching patterns reported in the literature. In this study, the potential of the biogenic ZnO NPs was explored for multifunctional applications. Zinc oxide nanoparticles exhibited a higher capacity for adsorbing hydrogen sulfide (H2S) compared to bulk zinc oxide, mostly because of their larger surface area. In addition, electrochemical studies demonstrated a substantial enhancement in the corrosion resistance of mild steel in a 1.0 M HCl solution. ZnO NPs also demonstrated remarkable photodegradation effectiveness, reducing 75% of methyl orange in 60 min under sun-light irradiation. This implies that they could be used to remediate organic pollutants (organic dyes) from wastewater