Phytochemicals and acute oral toxicity studies of the aqueous extract of Vernonia amygdalina from state of Malaysia

Vernonia amygdalina (VA) is a member of Asteracea family that use as traditional folk medicine in treating various infection and diseases. This study aim to determine the phytochemical characterization of Malaysian local VA and determine the acute oral toxicity. Phytochemical investigation was done by standard procedures. The crude extract of VA was observed to contained flavonoids, terpenoids, saponin and tannin. FTIR spectroscopy was done against VA aqueous extract revealed a presence of high content of flavonoids and terpenoids, phenols, methoxy compounds, ester carbonyl and amide. Sighting study for acute toxicity was conducted as per Organization of Economic Cooperation and Development (OECD) 425 guidelines which divided into sighting study and main study. One female Sprague Dawley rat was given single oral dose of VA aqueous extract at progressively dose of 175, 550, 2000 and 5000 mg/kg of body weight (BW), dissolved in 1ml distilled water and observed daily for 14 days. VA aqueous extract of 5000 mg/kg was used in main study as it exhibited no toxicity signs and mortality. All the treated rats survived and no toxicity signs were observed. No differences in body weights and organs weights. Liver function tests showed the levels of aspartate aminotransferase (AST) and globulin slightly increased but the levels of alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), albumin, total protein and total bilirubin were in normal range. Despite that, no elevation in the level of Alkaline phosphatase (ALP). Therefore, it can be concluded that the toxicity of VA is greater than 5000mg/kg BW as proved by AOT425 Statistical Programme

In vivo antidiabetic efficacy of Malaysian Vernonia amygdalina aqueous extract

Diabetes mellitus (DM) is a major world health concern especially DM types 2. In Malaysia, about 2.6 million adults were diagnosed with diabetes and most of them turn to herbal medicine for treatment. Vernonia amygdalina or locally known as bitter leaf is believed by the local people and traditional inhaler to reduce blood glucose level. This study aims to prove the antidiabetic property of Malaysian local V. amygdalina as claim by the local Malaysian. V. amygdalina aqueous extract was prepared by reflux method and was further concentrated. Twenty male Sprague Dawley rats were divided into 4 groups; non-diabetic rats as normal control (NC), diabetic induced rats as diabetic control (DC), diabetic induced rats treated with 150 mg/kg metformin and diabetic induced rats treated with 50 mg/kg V. amygdalina aqueous extract. The diabetic rats were induced by using 40 mg/kg of Streptozotocin (STZ). The study was conducted for 28 days. Body weights (BW) were taken at weekly interval for 4 weeks. Fasting blood glucose was measured in 3 day intervals. At the end of experiment, blood samples were collected for lipid profile test and insulin secretion assay. There were no significant differences in BW, relative organ weight (ROW) and no organs abnormalities were observed in all experimental groups. Significant decreases in triglycerides, total cholesterol and fasting blood glucose gave a prove evidence that V. amygdalina possessing antidiabetic property. However, the levels of insulin in diabetic induced rats treated with V. amygdalina aqueous extract were not significant as compared to the diabetic induced rats treated with metformin. The aqueous extract of V. amygdalina has antidiabetic activity. This work supports the folk use of this plant in treating diabetes. This antidiabetic property of Malaysian V. amygdalina may due to its phytochemical constituents as our previous study revealed the high contents of flavonoids and terpenoids. However, the aqueous extract of V. amygdalina did not act through regulation of insulin hormone since there was no significant change in insulin level

Cytotoxic effect of clinacanthus nutans semi-purified fraction (SF1) in combination with cisplatin against human cervical cancer

Cervical cancer is one of the most diagnosed malignancies in the world, and it is associated with HPV virus infection and invasion. A standard fraction from the medicinal plant Clinacanthus nutans was successfully separated, isolated, and identified as SF1 (semi-purified fraction). According to the previous study, SF1 has shown cytotoxic action against the cervical cancer cell line, SiHa. This study aims to further annotate the effect of SF1 in combination with cisplatin against SiHa to increase the selectivity of the anti-cancer treatment. The MTT assay was used to analyse the combination treatment’s cytotoxicity, and its index value was calculated using the Chou-Talalay method. A cell survival test was conducted to evaluate the reproducibility of cancer cells after being treated. The combination treatment has enhanced cytotoxic action and inhibited SiHa’s cell proliferation the most (IC50 value = 5.10 ± 0.86 μg/mL) compared to the individual cytotoxic activity of SF1 (IC50 value = 9.60 ± 0.20 µg/mL) and cisplatin (IC50 value = 3.60 ± 0.60 μg/mL). Simultaneously, the combination study has shown lesser cytotoxic activity towards normal cells, Vero (IC50 value = 10.30 ± 3.10 μg/mL) compared to SiHa cells. The combination also showed an antagonism effect with CI values of 2.60 to 1.50 and fractional inhibition (Fa) of 0.50 to 0.90. The findings have demonstrated that in contrast with single-agent therapy, the treatment of SF1 and cisplatin in combination has increased the efficacy because it selectively targets cancer cells by antagonism action

Malaysian herbs as potential natural resources of anticancer drugs: from folklore to discovery

Cancer is a complex disease and ranks as a leading cause of death globally. Despite many advances made in cancer therapeutics, adverse side effects and treatment resistance remain a great problem. In that sense, there are increasing demands to discover new anticancer agents from naturallyderived compounds. Medicinal plants represent a valuable source of new drugs with promising efficacy and safety. They produce various secondary metabolites, which exhibit unique structures and a pharmacological spectrum of activity, including antitumour activity. Clinacanthus nutans, Strobilanthes crispus, Ficus deltoidea, Curcuma longa, Centella asiatica and Piper betle are among the plants species commonly used to cure cancer in traditional medicine formulae in Malaysia. The present review aims to highlight the anticancer properties of the listed Malaysian herbs with a focus on their bioactive compounds and the mode of action. Overall, many studies have disclosed the presence of active metabolites in these plants, including phenols, alkaloids, flavonoids, terpenoids, saponin, curcumin and Asiatic acid. They possess significant cytotoxic or antiproliferative effects primarily via the induction of apoptosis, elevation of antioxidant activity and inhibition of cancer activating enzymes. Hence, further investigation into their clinical therapeutic potential may be noteworthy. Additionally, this review article also provides the reader with information concerning the conventional anticancer drugs and their limitations, recent developments and milestones achieved in plant- derived cancer therapeutics as well as different approaches to enhance the production of these anticancer molecules

Chemical profiling of volatile organic compounds from shoe odour for personal identification

Background: Body odour of an individual has a unique composition due to the combined influences of genetic, dietary, and environmental factors. This exploratory study was conducted to investigate the feasibility of obtaining unique chemical signatures of individuals from the shoe odour as a human chemical fingerprint for forensic identification. Methods: Five adult males and five adult females participated in this study and provided with two pairs of new canvas sports shoes. The participants were asked to do strenuous activities while wearing the shoes for 2 weeks. Two different extraction methods: swabbing technique, followed with direct extraction using different solvents, and air passive sampling technique, were compared for the effectiveness to extract the volatile organic compounds (VOCs) from the shoes. The variations of VOCs were investigated using gas chromatography coupled with flame ionisation detector (GC-FID), and the resultant chromatogram profiles collected from the shoe odour were further studied for individual identification purposes. In addition, principal component-discriminant analyses (PCA-DA) were then carried out on the GC dataset. Results: This study demonstrated that air passive sampling technique using methanol as the extraction solvent was effective for the detection of shoe odour. PCA-DA had successfully distinguished GC profile patterns and discriminated the shoe odour samples accordingly to their owners. The accuracy of DA for classification was 100%, with all samples were classified correctly to their groups. Conclusion: The chemical compounds of shoe odour have the potential to be utilised to aid the forensic investigation by identifying and narrowing down the possible suspect if shoes were recovered as evidence at the crime scene. © 2020, The Author(s)

SF1 : a standardised fraction of clinacanthus nutans that inhibits the stemness properties of cancer stem-like cells derived from cervical cancer

Cancer stem cells (CSCs) are a small population of tumour cells that are responsible for tumour initiation, metastases, recurrence, and resistance to conventional therapy. Hence, targeting CSCs is crucial in the fight against cancer. SF1, a standardised fraction from Clinacanthus nutans leaf extract, has been reported to exhibit potent and selective antineoplastic effects against cervical cancer cells. In this study, the potential of SF1 to inhibit the stemness of cervical cancer stem-like cells has been evaluated. SF1 extraction was carried out using the dry column vacuum chromatography technique. SiHa cell lines were cultured as spheres in CSC-conditioned medium (cervospheres), and the IC50 of SF1 against cervospheres was determined using the OZBlue Cell Viability Kit. The effects of SF1 on the cervosphere’s stemness markers, including CD49f, CK17, SOX2, OCT4, and NANOG, were assessed using a flow cytometry assay. Self-renewal inhibition and anti-tumorigenesis effects of SF1 in cervospheres were evaluated using a sphere formation assay and a xenograft mouse model. The present study shows that SF1 treatment at an IC50 of 17.07 µg/mL inhibited the proliferation, self-renewal, and tumorigenic capacity of SiHa cervospheres in vitro and in vivo. A decrease in the expressions of CK17, SOX2, CD49f, and OCT4 in cervical CSCs indicated that SF1’s inhibitory effects were also associated with the suppression of stemness markers. These results suggest that SF1 possesses an antitumor effect against cervical CSCs and may be regarded as a promising approach to the development of targeted anticancer agents for cervical cancer

SF1: a standardised fraction of Clinacanthus nutans that inhibits the stemness properties of cancer stem-like cells derived from cervical cancer = (SF1: fraksi piawaian Clinacanthus nutans yang merencat sifat stem sel menyerupai sel stem kanser diperolehi daripada kanser serviks)

Cancer stem cells (CSCs) are a small population of tumour cells that are responsible for tumour initiation, metastases, recurrence, and resistance to conventional therapy. Hence, targeting CSCs is crucial in the fight against cancer. SF1, a standardised fraction from Clinacanthus nutans leaf extract, has been reported to exhibit potent and selective antineoplastic effects against cervical cancer cells. In this study, the potential of SF1 to inhibit the stemness of cervical cancer stem-like cells has been evaluated. SF1 extraction was carried out using the dry column vacuum chromatography technique. SiHa cell lines were cultured as spheres in CSC-conditioned medium (cervospheres), and the IC50 of SF1 against cervospheres was determined using the OZBlue Cell Viability Kit. The effects of SF1 on the cervosphere’s stemness markers, including CD49f, CK17, SOX2, OCT4, and NANOG, were assessed using a flow cytometry assay. Self-renewal inhibition and anti-tumorigenesis effects of SF1 in cervospheres were evaluated using a sphere formation assay and a xenograft mouse model. The present study shows that SF1 treatment at an IC50 of 17.07 µg/mL inhibited the proliferation, self-renewal, and tumorigenic capacity of SiHa cervospheres in vitro and in vivo. A decrease in the expressions of CK17, SOX2, CD49f, and OCT4 in cervical CSCs indicated that SF1’s inhibitory effects were also associated with the suppression of stemness markers. These results suggest that SF1 possesses an antitumor effect against cervical CSCs and may be regarded as a promising approach to the development of targeted anticancer agents for cervical cancer

Evaluation of acute and sub-acute oral toxicity of the aqueous extract of aquilaria malaccensis leaves in sprague dawley rats

Aquilaria malaccensis or commonly known as ‘gaharu’ is a species of Aquilaria genus and belongs to the Thymelaeaceae family. It is widely distributed in Malaysia, Indonesia, and the Borneo Islands. Traditionally, its leaves were used to relieve bruises and studies have shown that they function as an antioxidant, aphrodisiac, and tranquilizer. Despite its proven beneficial medicinal properties, information regarding its toxicity is limited. Therefore, we performed a safety evaluation on the aqueous A. malaccensis leaves extract (AMAE) in Sprague Dawley rats. The assessment of acute toxicity based on the Organization for Economic Cooperation and Development (OECD) Guideline 420 revealed that AMAE did not influence mortality, clinical appearance, body weight gain, or necropsy findings at a dose of 2000 mg/kg body weight. In the sub-acute toxicity, all doses did not significantly modify the body weight and food and water intake. In male rats treated with 2000 mg/kg, there was a significant reduction in the relative weight of liver. Not only that, an increase in alkaline phosphatase and alanine transaminase was also observed in different groups among the female rats. A significant decrease in the creatinine level was also seen among male rats administered with different doses of AMAE. In both sexes, histopathological analysis had shown abnormalities in the liver and kidney of rats treated at the dose of 2000 mg/kg. In conclusion, the 50% lethal dose (LD50) of AMAE was estimated to be greater than 2000 mg/kg. In sub-acute duration, the findings suggested that AMAE administered orally is slightly toxic at higher doses (2000 mg/kg) and could provoke functional and structural changes in the kidney and liver of rats. Thus, the extract should be used with caution

Nanocarbon powder for latent fingermark development: a green chemistry approach

Abstract Background Optimal fingermark developing product would have the following attributes; small, rounded and well-dispersed particles, which has high affinity for the fingermark residue but exhibits minimal interaction with the substrate bearing the fingermark. However, most commercial powder formulations still utilise fine particles in the range of 1 to 10 μm. Results Ageing of the filtrate from rice husk acid digestion produced amorphous nanocarbon powder (NP) that exhibited non-smooth, slightly irregular spherical particles with a higher degree of agglomeration. Particle size distribution fell in the range of 100 to 500 nm with mean particle size of 300 nm. Molecular bonding and chemical composition of the NP was characterised using spectroscopic techniques. Findings revealed that the nanoparticle powders and reagent performed on par with the existing commercial powders and exhibited higher selectivity. Nanoparticle synthesis from low-cost precursor in this research did not require special equipment, addition of resins or adhesives, or surface passivation. Conclusion Nanocarbon powder is exploitable as fingermark developing powder generated from a sustainable source

Semi-purified fraction of Clinacanthus nutans induced apoptosis in human cervical cancer, SiHa cells via up-regulation of bax and down-regulation of Bcl-2

Cervical cancer is a leading cause of death in women, worldwide, and second in Malaysia. Nowadays, modern cancer treatment comes with negative and adverse side effects to the patients. Traditional medicinal plants such as Clinacanthus nutans has been locally recognized for its medicinal properties and was claimed for cancer treatment. According to our pilot study, a fraction of C. nutans has shown a potent inhibition on human cervical cancer cells, SiHa, in vitro. Hence, this study aimed to investigate the underlying anticancer mechanism of a semi-purified fraction (SF1) of C. nutans towards cervical cancer cells. Bioassay guided fractionation and further purification was carried out. All collected fractions of C. nutans were examined for cytotoxicity activity towards HeLa, SiHa and NIH cells by MTT Assay. SF1 exhibited better cytotoxic activity with best growth inhibition against SiHa cells (IC50 value = 9.60 μg/mL). Cytoselective effects were shown as no cytotoxicity observed in the treated normal NIH cells. SF1 induced early apoptosis event in SiHa cells as resulted by FITC-Annexin V/propidium iodide staining by flowcytometric analysis. The protein expression by flowcytometry using specific antibody conjugated fluorescent dye showed up-regulation of Bax and down-regulation Bcl-2 as well as increment of cytochrome C levels upon treatment with SF1. In conclusion, the findings suggested that SF1 demonstrated antiproliferative effects and induced apoptosis in SiHa cells through up-regulation of Bax and down-regulation of Bcl-2 via mitochondrial-dependent pathway