MyMedR

Fulltext Cloning and expression of a subfamily 1.4 lipase from Bacillus licheniformis IBRL-CHS2

Reddy, Nidyaletchmy Subba, Rashidah Abdul Rahim, Darah Ibrahim, Kumar, K. Sudesh

Trop Life Sci Res, 2016;27(11):145-150.

MyJurnal

We report on the cloning of the lipase gene from Bacillus licheniformis IBRLCHS2
and the expression of the recombinant lipase. DNA sequencing analysis of the
cloned lipase gene showed that it shares 99% identity with the lipase gene from
B. licheniformis ATCC 14580 and belongs to subfamily 1.4 of true lipases based on amino
acid sequence alignment of various Bacillus lipases. The 612 bp lipase gene was then
cloned into the pET-15b(+) expression vector and the construct was transformed into
E. coli BL21 (DE3) for bulk expression of the lipase. Expression was analysed by SDSPAGE
where the lipase was found to have a molecular weight of about 23 kDa.

In vitro antimicrobial activity against Pseudomonas aeruginosa and acute oral toxicity of marine algae Gracilaria changii

Sasidharan S, Darah I, Noordin MK

N Biotechnol, 2010 Sep 30;27(4):390-6.

PMID: 20170762 DOI: 10.1016/j.nbt.2010.02.002

Methanol extract of the Gracilaria changii has been screened for antimicrobial activity against Pseudomonas aeruginosa. Antimicrobial activities were carried out using disc diffusion assay and broth dilution method against P. aeruginosa. The methanol extract of G. changii showed a good antimicrobial activity against P. aeruginosa with MIC (Minimum Inhibitory Concentration) value of 6.25mg/ml. Exposure of P. aeruginosa cells to 6.25mg/ml of methanol extract of G. changii resulted in complete inhibition of the bacterial cells. The main abnormalities noted via SEM and TEM studies were the alterations in morphology and cytology of the bacterial cells. The main reason for this deterioration was discussed. The effect of the methanol extract on the growth profile for the bacteria was also done and confirmed the bactericidal effect of the G. changii methanol extract on P. aeruginosa by changing the normal growth profile of P. aeruginosa. In an acute toxicity study using mice, the median lethal dose (LD(50)) of the extract was greater than 2000 mg/kg, and we found no pathological changes in macroscopic examination by necropsy of mice treated with extract. We conclude that G. changii might be safely used as an antimicrobial agent.

Fulltext Preliminary isolation and in vitro antiyeast activity of active fraction from crude extract of Gracilaria changii

Sasidharan S, Darah I, Noordin MK

Indian J Pharmacol, 2008 Oct;40(5):227-9.

PMID: 20040962 DOI: 10.4103/0253-7613.44155

OBJECTIVE: To isolate the active fraction from crude extract of Gracilaria changii and to determine its in vitro antifungal activity.

MATERIALS AND METHODS: The active fraction was isolated from the crude extract of G. changii by various purification procedures such as column chromatography, thin layer chromatography, bioauthograph etc. The in vitro antifungal activity (Candida albicans) of the active fraction (1.00, 0.50, and 0.25 mg/ml) was studied by disc diffusion method and the effect of the active fraction on the morphology of yeast was done by scanning electron microscope (SEM) studies.

RESULTS: An active fraction with remarkable antifungal activity was separated from the crude extract. The active fraction was effective as a fungicide against C. albicans and showed a dose-dependent antifungal activity. A Scanning Electron Microscope (SEM) study confirmed the fungicidal effect of G. changii active fraction on C. albicans, by changing the normal morphology of C. albicans.

CONCLUSION: From G. changii crude extract, an active fraction with remarkable in vitro antifungal activity has been isolated.

Fulltext Acute and subchronic toxicity study of Euphorbia hirta L. methanol extract in rats

Yuet Ping K, Darah I, Chen Y, Sreeramanan S, Sasidharan S

Biomed Res Int, 2013;2013:182064.

PMID: 24386634 DOI: 10.1155/2013/182064

Despite Euphorbia hirta L. ethnomedicinal benefits, very few studies have described the potential toxicity. The aim of the present study was to evaluate the in vivo toxicity of methanolic extracts of E. hirta. The acute and subchronic oral toxicity of E. hirta was evaluated in Sprague Dawley rats. The extract at a single dose of 5,000 mg/kg did not produce treatment related signs of toxicity or mortality in any of the animals tested during the 14-day observation period. Therefore, the LD 50 of this plant was estimated to be more than 5,000 mg/kg. In the repeated dose 90-day oral toxicity study, the administration of 50 mg/kg, 250 mg/kg, and 1,000 mg/kg/day of E. hirta extract per body weight revealed no significant difference (P > 0.05) in food and water consumptions, body weight change, haematological and biochemical parameters, relative organ weights, and gross findings compared to the control group. Macropathology and histopathology examinations of all organs including the liver did not reveal morphological alteration. Analyses of these results with the information of signs, behaviour, and health monitoring could lead to the conclusion that the long-term oral administration of E. hirta extract for 90 days does not cause sub-chronic toxicity.

Fulltext Cytotoxicity and genotoxicity assessment of Euphorbia hirta in MCF-7 cell line model using comet assay

Ping KY, Darah I, Chen Y, Sasidharan S

Asian Pac J Trop Biomed, 2013 Sep;3(9):692-6.

PMID: 23998008 DOI: 10.1016/S2221-1691(13)60140-9

To evaluate the cytotoxicity and genotoxicity activity of Euphorbia hirta (E. hirta) in MCF-7 cell line model using comet assay.

Fulltext Genotoxicity of Euphorbia hirta: an Allium cepa assay

Yuet Ping K, Darah I, Yusuf UK, Yeng C, Sasidharan S

Molecules, 2012 Jun 26;17(7):7782-91.

PMID: 22735780 DOI: 10.3390/molecules17077782

The potential genotoxic effects of methanolic extracts of Euphorbia hirta which is commonly used in traditional medicine to treat a variety of diseased conditions including asthma, coughs, diarrhea and dysentery was investigated using Allium cepa assay. The extracts of 125, 250, 500 and 1,000 µg/mL were tested on root meristems of A. cepa. Ethylmethanesulfonate was used as positive control and distilled water was used as negative control. The result showed that mitotic index decreased as the concentrations of E. hirta extract increased. A dose-dependent increase of chromosome aberrations was also observed. Abnormalities scored were stickiness, c-mitosis, bridges and vagrant chromosomes. Micronucleated cells were also observed at interphase. Result of this study confirmed that the methanol extracts of E. hirta exerted significant genotoxic and mitodepressive effects at 1,000 µg/mL.

Antibacterial activity and morphological changes of Pseudomonas aeruginosa cells after exposure to Vernonia cinerea extract

Latha LY, Darah I, Kassim MJ, Sasidharan S

Ultrastruct Pathol, 2010 Aug;34(4):219-25.

PMID: 20594042 DOI: 10.3109/01913121003651513

The antibacterial activity of Vernonia cinerea (L.) extract was investigated using the broth dilution method. The extract showed a favorable antimicrobial activity against Pseudomonas aeruginosa with a minimum inhibition concentration (MIC) value of 3.13 mg/mL. V. cinerea extract at (1/2), 1, or 2 times the MIC significantly inhibited bacterial growth with a noticeable drop in optical density (OD) of the bacterial culture, thus confirming the antibacterial activity of the extract on P. aeruginosa. Imaging using scanning (SEM) and transmission (TEM) electron microscopy was done to determine the major alterations in the microstructure of the extract-treated P. aeruginosa. The main abnormalities noted via SEM and TEM studies were the alteration in morphology of the bacterial cells. The main reason for this destruction was the severe alterations of the cell wall with the formation of holes, invaginations, and morphological disorganization caused by the extract. The authors conclude that the extract may be used as a candidate for the development of antimicrobial agents.

Fulltext Pharmacological screening of methanolic extract of Ixora species

Latha LY, Darah I, Jain K, Sasidharan S

Asian Pac J Trop Biomed, 2012 Feb;2(2):149-51.

PMID: 23569886 DOI: 10.1016/S2221-1691(11)60210-4

OBJECTIVE: To investigate the antimicrobial activity of methanolic extracts of different parts of Ixora species.

METHODS: Antimicrobial activity was carried out using disc diffusion assay against fungi, gram-positive and gram-negative bacteria.

RESULTS: All methanolic extracts of different parts of Ixora species showed a broad-spectrum of antibacterial and antiyeast activities, which inhibited the growth of at least one bacterium or yeast. There was no remarkable difference between different Ixora species observed in this study.

CONCLUSIONS: The significant antimicrobial activity shown by this Ixora species suggests its potential against infections caused by pathogens. The extract may be developed as an antimicrobial agent.

Fulltext Identification of small molecule inhibitors of p27(Kip1) ubiquitination by high-throughput screening

Ooi LC, Watanabe N, Futamura Y, Sulaiman SF, Darah I, Osada H

Cancer Sci, 2013 Nov;104(11):1461-7.

PMID: 23910095 DOI: 10.1111/cas.12246

Dysregulation of p27(Kip1) due to proteolysis that involves the ubiquitin ligase (SCF) complex with S-phase kinase-associated protein 2 (Skp2) as the substrate-recognition component (SCF(Skp2)) frequently results in tumorigenesis. In this report, we developed a high-throughput screening system to identify small-molecule inhibitors of p27(Kip1) degradation. This system was established by tagging Skp2 with fluorescent monomeric Azami Green (mAG) and CDK subunit 1 (Cks1) (mAGSkp2-Cks1) to bind to p27(Kip1) phosphopeptides. We identified two compounds that inhibited the interaction between mAGSkp2-Cks1 and p27(Kip1): linichlorin A and gentian violet. Further studies have shown that the compounds inhibit the ubiquitination of p27(Kip1) in vitro as well as p27(Kip1) degradation in HeLa cells. Notably, both compounds exhibited preferential antiproliferative activity against HeLa and tsFT210 cells compared with NIH3T3 cells and delayed the G1 phase progression in tsFT210 cells. Our approach indicates a potential strategy for restoring p27(Kip1) levels in human cancers.

In vitro antioxidant activity potential of lantadene A, a pentacyclic triterpenoid of Lantana plants

Grace-Lynn C, Darah I, Chen Y, Latha LY, Jothy SL, Sasidharan S

Molecules, 2012 Sep 19;17(9):11185-98.

PMID: 22992785

Lantadenes are pentacyclic triterpenoids present in the leaves of the plant Lantana camara. In the present study, in vitro antioxidant activity and free radical scavenging capacity of lantadene A was evaluated using established in vitro models such as ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picryl-hydrazyl (DPPH•), hydroxyl radical (OH•), nitric oxide radical (NO•), superoxide anion scavenging activities and ferrous ion chelating assay. Interestingly, lantadene A showed considerable in vitro antioxidant, free radical scavenging capacity activities in a dose dependant manner when compared with the standard antioxidant in nitric oxide scavenging, superoxide anion radical scavenging and ferrous ion chelating assay. These findings show that the lantadene A possesses antioxidant activity with different mechanism of actions towards the different free radicals tested. Since lantadene A is a very popular drug in modern medicine, it is a promising candidate for use as an antioxidant and hepatoprotective agent.

Fulltext Cassia spectabilis (DC) Irwin et Barn: a promising traditional herb in health improvement

Jothy SL, Torey A, Darah I, Choong YS, Saravanan D, Chen Y, et al.

Molecules, 2012 Aug 29;17(9):10292-305.

PMID: 22932211 DOI: 10.3390/molecules170910292

The genus Cassia, comprising about 600 species widely distributed worldwide is well known for its diverse biological and pharmacological properties. Cassia spectabilis (sin Senna spectabilis) (DC) Irwin et Barn (Fabaceae) is widely grown as an ornamental plant in tropical and subtropical areas. C. spectabilis has been commonly used in traditional medicine for many years. Information in the biomedical literature has indicated the presence of a variety of medicinally-important chemical constituents in C. spectabilis. Pharmacological studies by various groups of investigators have shown that C. spectabilis possesses significant biological activity, such as antibacterial, antibiofilm, antifungal and antioxidant properties. Beside this, toxicity studies of this plant have revealed no toxic effect on mice. In view of the immense medicinal importance of C. spectabilis, this review aimed at compiling all currently available information on C. spectabilis’s botany, phytochemistry, pharmacology, and mechanism of actions, toxicology and its ethnomedicinal uses.

Displaying all 11 publications