Effects of epigallocatechin-3-gallate (EGCG) on cell cycle distribution and DNA integrity of K562 cells, a human chronic myeloid leukemia

Epigallocatechin-3-gallate (EGCG) is a naturally derived compound from green tea with high antioxidant activity and various anti-cancer properties. EGCG has been widely investigated worldwide. However, effects of EGCG on cell cycle of K562 have not been clearly stated elsewhere. This study was conducted with the aim to investigate the antiproliferative effect of EGCG on K562 human leukemic cells and its underlying mechanism of action on the cells. MTT assay was conducted to determine cytotoxicity effect of EGCG on the K562 cells. Meanwhile, cell cycle analysis and DNA damage on the cells were determined by Flow cytometry and Comet assay respectively. K562 cells were treated with EGCG at concentrations ranging from 0 to 100μg/ml for 48 hours. The results showed that EGCG effectively decreased the percentage of cell viability in a dose dependent manner. The IC10, IC25 and IC50 of EGCG on K562 cell lines were 5 ± 2.44 μg/mL, 10 ± 5.93 μg/mL and 50 ± 1.93 μg/mL, respectively. In cell cycle assay, EGCG has shown no significant effect (p>0.05) on the cell cycle of K562 cell line as compared to negative control, whereas Imatinib mesylate as the positive control showed cell cycle arrest at S phase in this cell line. Hence, EGCG can be verified as a non-cell cycle specific compound. In addition, EGCG was found to cause a significant increase (p<0.05) in tail moment value and percentage of DNA tail in K562 cell line, suggesting DNA damage as an early signal of EGCG induced cell cytotoxicity. In conclusion, by decreasing the cell viability and inducing DNA damage, EGCG showed promising potential as an alternative treatment for leukemia through non-cell cycle specific pathway and further investigation on other mechanisms of action of EGCG on the cells is recommended

Protective effects of Zingiber zerumbet ethyl acetate extract on hydrogen peroxide-induced damage of red blood cells

Zingiber zerumbet is widely used as therapeutic agent in traditional medicine and is reported to exert antioxidant activities. This study evaluated in vitro antioxidant potential of Z. zerumbet rhizome ethyl acetate extract on hydrogen peroxide (H2O2)-induced red blood cells (RBCs) damage; by measuring percentage of haemolysis, oxidative damage and morphological changes in treated RBCs. On a preliminary dose-response analysis, it was observed that incubation of RBCs with extract at doses of 50, 25, 12.5 and 6.25 μg/mL all had no significant effect on RBCs similar to NaCl control and BHT (positive control), therefore, these concentrations were used in the subsequent study. The RBCs were pre-incubated with extract in the chosen concentrations, prior to treatment with H2O2. The results showed that only 6.25 μg/mL+H2O2 group showed significantly (p<0.05) lower percentage of haemolysis and oxidative damage; indicated by low level of malondialdehyde (MDA) and protein carbonyls (PC) compared to the H2O2 alone group. Electron microscopic examination further showed that pre-treatment with 6.25 μg/mL extract reduced the H2O2 –induced morphological changes of RBCs. Phytochemical analysis of extract using Gas Chromatography Mass Spectrometry (GCMS) identified Zerumbone as the highest constituent compound (51.57%) in this extract. In conclusion, this study indicated that 6.25 μg/mL ethyl acetate extract Z. zerumbet rhizome could efficiently protect RBCs against oxidative damage induced by H2O2 and this effects could possibly due the high constituent of Zerumbone

Neuroprotective effects of ethyl acetate extract of Zingiber zerumbet (L.) Smith against oxidative stress on paraquat-induced parkinsonism in rats

Zingiber zerumbet has been traditionally used as an anti-inflammation and antioxidant agent. The present study investigates the neuroprotective effects of ethyl acetate extract of Z. zerumbet against oxidative stress on paraquat (PQ)-induced Parkinsonism in rats. Forty male Sprague-Dawley rats were divided into five groups: Negative control (normal saline), positive control (N-acetylcysteine, NAC 20 mg/kg + PQ 10 mg/kg), PQ only, 200 mg/kg Z. zerumbet + PQ and 400 mg/kg Z. zerumbet + PQ. The extract was given orally for 19 consecutive days and PQ was administered intraperitoneally on day 8-12th of the treatment regime. Both serum and fresh brains containing substantia nigra (SN) region were taken for biochemical and histological analysis. Administration of both 200 and 400 mg/kg ethyl acetate Z. zerumbet extracts to the PQ-treated groups have resulted in: Decreased levels of MDA and PC in the SN homogenates; and increased SOD, GPx; and CAT activities in the SN and serum. Overall, ethyl acetate extract of Z. zerumbet reduced oxidative stress in the SN of PQ-induced neuronal damages, therefore, has the potential to be developed as a preventive agent for neurodegenerative disorders caused by environmental toxins

Supercritical fluid extraction of bioactive flavonoid from Strobilanthes crispus (pecah kaca) and its comparison with solvent extraction

Supercritical carbon dioxide extraction (SC-CO 2) of bioactive flavonoid from Strobilanthes crispus (Pecah Kaca) was performed to study the effects of various parameters such as pressure, temperature and dynamic extraction time on the yield and composition of bioactive flavonoid. The results were also compared with those obtained by conventional Soxhlet extraction in lab conditions. The results from SFE showed that the effect of extraction variables on extraction yields decreased in the following order: pressure, temperature and dynamic extraction time. The extraction pressure played a dominant role in the yield of the sample while the effect of time could be ignored. This study also revealed that both Soxhlet extraction and SC-CO 2 extraction can be used to obtain flavonoid compound. Under the optimum conditions, the highest bioactive flavonoid compound content was obtained at 3.98% and eight flavonoid compounds were identified by HPLC

Herbal Formulation C168 Attenuates Proliferation and Induces Apoptosis in HCT 116 Human Colorectal Carcinoma Cells: Role of Oxidative Stress and DNA Damage

The use of herbal formulations has gained scientific interest, particularly in cancer treatment. In this study, the herbal formulation of interest, denoted as C168, is a mixture of eight genera of plants. This study aims to investigate the antiproliferative effect of C168 methanol extract (CME) on various cancer cells and its underlying mechanism of action on the most responsive cell line, namely, HCT 116 cells. CME exerted antiproliferative activities on HCT 116 colorectal carcinoma cells and HepG2 hepatocellular carcinoma cells but not on CCD-841-CoN normal colon epithelial cells, Jurkat E6.1 lymphoblastic leukemic cells, and V79-4 Chinese hamster lung fibroblasts. Further investigation on HCT 116 cells showed that CME induced G2/M cell-cycle arrest and apoptosis. Treatment of CME induced oxidative stress in HCT 116 cells by increasing the superoxide anion level and decreasing the intracellular glutathione. CME also increased tail moment value and H2AX phosphorylation in HCT 116 cells, suggesting DNA damage as an early signal of CME induced apoptosis. Loss of mitochondrial membrane potential in CME-treated cells also indicated the involvement of mitochondria in CME induced apoptosis. This study indicated the selectivity of CME toward colon cancer cells with the involvement of oxidative damage as its possible mechanism of action

Zingiber zerumbet (L.) Smith hexane crude extract caused DNA damage on Leptospira spp.

Numerous attempts have been made to control leptospirosis by using chemoprophylaxis, but with limited success. The present study was done to investigate the antileptospiral potential of hexane, ethyl acetate and methanol extracts of Zingiber zerumbet rhizomes. The extracts were assayed for antileptospiral activity using broth microdilution method against Leptospira interrogans (serovar Batavie, Canicola, Australis) and Leptospira biflexa serovar Patoc. The Z. zerumbet hexane extract exhibited antileptospiral activity, with IC50 values of 248 μg/mL against L. interrogans serovar Canicola, IC50 of 125 μg/mL against L. interrogans serovar Australis, IC50 of 15.63 μg/mL against L. interrogans serovar Batavie and IC50 of 109 μg/mL against L. biflexa serovar Patoc. However, both ethyl acetate and methanol extracts did not show any distinct antileptospiral activity. Since the hexane extract of Z. zerumbet showed antileptospiral activity, the DNA-damaging properties of this extract were tested according to their IC50 and IC25 values that were specific to each serovars. The DNA-damaging properties were determined by treating the selected Leptospira spp. with the hexane extract and subjecting its DNA to electrophoresis and analysis on agarose gels. The results demonstrated that the hexane extract had DNA-damaging properties towards L. biflexa serovar Patoc and L. interrogans serovar Australis, as proven by the appearance of fragmented DNA on the gels. We conclude that the Z. zerumbet hexane extract could inhibit the growth of Leptospira spp. serovar Patoc and Australis through DNA-damaging activity and thus, could be a potential antileptospiral agent. Further studies are needed to investigate the potential of this hexane extract as an antileptospiral agent using in vivo rat models of leptospirosis

Analisa sitogenetik sel bukal petani di Tanjung Karang dan Kelantan yang terdedah kepada pestisid

Pestisid dan baja kimia digunakan secara meluas dalam sektor pertanian bagi meningkatkan hasil pertanian dalam kalangan petani. Namun, pendedahan kepada pestisid akan memberi potensi risiko kepada kesihatan manusia. Kajian ini bertujuan menganalisa kekerapan pembentukan mikronukleus (MN) dan binukleus (BNu) pada mukosa sel bukal petani yang terdedah kepada pestisid dengan menggunakan asai MN. Perbandingan kekerapan MN dan Bnu dilakukan di dua kawasan iaitu Tanjung Karang, Selangor dan Kelantan kerana aktiviti pertanian dan jenis pestisid yang digunakan adalah berbeza. Pengambilan sel bukal dilakukan pada petani di Tanjung Karang (n = 32) dan petani di Kelantan (n = 43) dengan mnggunakan kayu penyendal lidah. Borang soal selidik juga digunakan untuk mendapatkan data demografik para petani. Analisa sitogenetik dilakukan dengan menggunakan pewarnaan Akridin Jingga (AO) 0.0025% (w/v). Kekerapan MN dan BNu yang terbentuk melalui analisa dibawah mikroskop fluoresen dijadikan sebagai petunjuk kerosakan sitogenetik. Keputusan kajian menunjukkan kekerapan MN dan BNu petani di Tanjung Karang adalah lebih tinggi secara signifikan (p 0.05) dan amalan pemakaian PPE (Personal Protective Equipment) (p > 0.05). Selain itu, ujian korelasi yang dijalankan menunjukkan terdapat korelasi positif antara kekerapan MN dengan tempoh pendedahan pestisid petani di Tanjung Karang (p > 0.05, r = 0.015) dan Kelantan (p > 0.05, r = 0.0158). Manakala, kekerapan BNu juga mempunyai korelasi positif dengan pendedahan pestisid petani di Tanjung Karang (p > 0.05, r = 0.036) dan petani di Kelantan (p > 0.05, r = 0.013). Justeru, kajian ini membuktikan bahawa pendedahan pestisid boleh meningkatkan pembentukan MN dan BNu dalam kalangan petani dan ini menjelaskan bahawa penggunaan pestisid dalam jangka masa panjang boleh mengaruh genotoksisiti dan kerosakan DNA kepada manusia

Preventive effects of Polygonum minus essential oil on cisplatin-induced hepatotoxicity in sprague dawley rats

Cisplatin is a chemotherapeutic agent widely used in treating various types of cancer. However, its usage is restricted due to the adverse hepatoxicity, as seen in approximately 36% of cancer patients receiving cisplatin treatment. Polygonum minus essential oil has high antioxidant capacity, and is enriched with terpenoids and phenolic compounds. The objective of this study was to investigate effects of P. minus essential oil (PmEO) supplementation on cisplatin-induced hepatotoxicity in rats. Male rats were divided into seven different groups, namely: control (C), cisplatin-induced (CP), positive control with β-caryophyllene 150 mg/kg (BCP), PmEO 100 mg/kg (PmEO100CP), PmEO 200 mg/kg (PmEO200CP), PmEO 400 mg/kg (PmEO400CP) and PmEO 400 mg/kg alone (PmEO400). PmEO and BCP was given orally for 14 days prior to a single dose cisplatin (10 mg/kg) injection on day 15 and rats were sacrificed on day 18. Liver enzymes, histology, ultrastructural morphology and oxidative stress markers such as glutathione, glutathione peroxidase, catalase, superoxide dismutase and malondialdehyde were assayed. Compared to controls, levels of transaminase enzymes, serum bilirubin and oxidative stress were all increased in CP, PmEO200CP and PmEO400CP groups. However, only PmEO100CP and BCP groups reduced these increases in level of transaminase enzymes and oxidative stress compared to CP group. On both light microscopic and ultrastructural examination, CP and PmEO400CP groups showed hepatotoxicity, exhibited by cytoplasmic vacuolation, congested blood sinusoids and increased number of Kupffer cells. However, these changes were minimized in the PmEO100CP group. Therefore, we concluded that PmEO given at 100 mg/kg has preventive effect against cisplatin-induced hepatotoxicity in rats

Polygonum minus essential oil modulates cisplatin-induced hepatotoxicity through inflammatory and apoptotic pathways

Oxidative stress, inflammation and apoptosis are thought as primary mediators of cisplatin-induced hepatotoxicity. The objective of this study was to determine the protective effect of Polygonum minus essential oil in cisplatin-induced hepatotoxicity. A total of forty-two male rats were randomly divided into seven groups: control, cisplatin, β-caryophyllene 150 mg/kg (BCP), PmEO 100 mg/kg + cisplatin (PmEO100CP), PmEO 200 mg/kg + cisplatin (PmEO200CP), PmEO 400 mg/kg + cisplatin (PmEO400CP) and PmEO 400 mg/kg (PmEO400). Rats in the BCP, PmEO100CP, PmEO200CP, PmEO400CP and PmEO400 group received respective treatment orally for 14 consecutive days prior to cisplatin injection. All animals except for those in the control group and PmEO400 were administered with a single dose of cisplatin (10 mg/kg) intraperitoneally on day 15 and all animals were sacrificed on day 18. PmEO100CP pretreatment protected against cisplatin-induced hepatotoxicity by decreasing CYP2E1 and indicators of oxidative stress including malondialdehyde, 8-OHdG and protein carbonyl which was accompanied by increased antioxidant status (glutathione, glutathione peroxidase, superoxide dismutase and catalase) as compared to cisplatin group. PmEO100CP pretreatment also modulated changes in liver inflammatory markers (TNF-α, IL-1α, IL-1β, IL-6 and IL-10). PmEO100CP administration also notably reduced cisplatin-induced apoptosis significantly as compared to cisplatin group. In conclusion, our results suggested that P. minus essential oil at a dose of 100 mg/kg may protect against cisplatin-induced hepatotoxicity possibly via inhibition of oxidative stress, inflammation and apoptosis

The effects of genetic inheritance E-game to undergraduate students

The COVID-19 outbreak that is occurring worldwide has affected the education system, as the closure of education institutions has halted teaching and learning activities. Nevertheless, online education is being widely applied to complete the curriculum. As learning activities are limited by the prohibition of face-to-face classes, an E-game on genetic disease inheritance was designed and introduced to undergraduate Biomedical Science students of Universiti Kebangsaan Malaysia (UKM). In this approach, the students were divided into small groups of 5-7 students and they were required to grasp the basic knowledge of genetic disease inheritance including drawing a pedigree tree in order to understand genetic-based diseases, develop three case studies, four case studies-related questions and also provide the answer scheme beforehand. During the E-game, the Microsoft Teams online learning platform was used. Each of the groups took turns to present case studies on the topic of genetic disease inheritance and answered questions. Marks were given for each of the answers. The students’ feedback was collected to evaluate the outcome of this approach. Their understanding on pedigree drawing topics, ability to draw pedigree diagram and analysing genetic inheritance case studies were significantly (p<0.001) increased after the e-game implementation. In addition, the e-game helped them to develop several skills such as teamwork and strategizing. Overall, the students were benefited with the approach, even though some limitations in internet accessibility and devices problems were encountered during the e-game. As conclusion, the e-game was successfully tailored to deliver the genetic inheritance topic previously outlined for face to face session