Biological Activitivies Of Acanthophora Spicifera (VAHL) Borgesen From Pulau Gedung, Penang

A total of eight extract of Acanthophora spicifera extracted using two different methods using solvents of different polarity, were used throughout this study. These extracts were assayed to determine the antimicrobial activities, antioxidant activities, in-vitro cytotoxicity and total phenolic contents. Most of the extracts exhibited antibacterial activity against the tested bacteria. Meanwhile, no antifungal activity was observed on tested fungi. Among the extract, crude methanol extract demonstrated promising antibacterial activity against Gram-negative Pseudomonas aeruginosa ATCC 27853 with inhibition zone of 11.0 mm, MIC values of 250 μg mL-1 and MBC value of 500 μg mL-1. A. spicifera also exhibited antioxidant properties

Studies of glucose-6-phosphate dehydrogenase

Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme that catalyzes the conversion of glucose-6-phosphate to 6-phosphoglucolactone with the reduction of NADP+ to NADPH. It is the first component of the pentose phosphate pathway (PPP) that is a central and important biochemical pathway. The PPP functions to produce building block compounds and intermediates for energy production. The first three enzymes of the PPP are referred to as the oxidative phase and produce NADPH that is crucial for dealing with oxygen stress, especially in people with G6PD deficiency. There are tests for G6PD deficiency, but those used in the field are not reliable. The existing assays for G6PD use the NADPH it generates to reduce a dye and produce a colour change, which can be observed with the naked eye and can be used to measure the kinetic parameters of the purified G6PD in the laboratory. Unfortunately, it suffers from a number of problems when used with crude preparations under field conditions. The equilibrium constant for the G6PD reaction is close to 1 so that the G6PD substrate will only be depleted if subsequent enzymes in the PPP are present. The extent of colour change will also depend upon the activity of the next two enzymes in the PPP and these may vary from person to person. The mutant forms of G6PD may be unstable, and may gives rise to false positive results during the assays. Hence, sensitive and reliable assays are needed. Here, we report an improvised colorimetric method for G6PD detection using tetrazolium salt, WST-8. The method used enzyme-coupling reaction of G6PD with another two downstream enzymes in the PPP. Addition of large excess of the two enzymes decreased the response time and increases the sensitivity of the test. Consequently, this enables detection of G6PD enzyme activity at low substrate concentrations compared with an assay of NADPH absorbance at 340 nm. The methods also generate NADPH eight times faster than reaction with G6PD alone. This makes the test more sensitive and predictive, which is also ideal for industrial applications. We also extend the application of the colorimetric WST-8 assay for screening of recombinant libraries in a directed evolution study for improved stability. The screening protocols were modified to colony agar-blotting using filter paper as primary screening and 96 well plates for secondary screening. The orange coloured formazans produced by the reaction, make it suitable and reliable for high-throughput qualitative and quantitative assays for dehydrogenases. With only four rounds of evolution, the method successfully identified promising variants with far improved thermal and chemical (urea) stability. Studies characterizing the properties and understanding the effect of mutations on variants are also discussed. The mutations responsible for improved stability were found located at the large structural NADP+ domain that is important for integrity and stability of the structure. The results also showed that the mutations brought changes in the oligomeric forms of G6PD. The thermostable variants were found to form higher order oligomers compared to the native enzyme. The correlation between enzyme stability and oligomerisation was also studied briefly

Bioactivity of clitoria ternatea crude extracts against pathogenic bacteria

Clitoria ternatea, sometimes referred to as the Asian pigeon wings blue pea, the butterfly pea, or the Darwin pea, is a Fabaceae plant species that has been shown to possess antibacterial effects against several pathogenic microbes. Hence, the present study has been carried out to access the antibacterial activity of C. ternatea flower extracted with water and methanol against pathogenic bacteria. The well and disk diffusion assays were performed to determine the antibacterial activity of C. ternatea flower extracts. The efficacy of the extracts was then evaluated via broth microdilution assay to obtain MIC and MBC values and the growth reduction assay. Meanwhile, the DPPH scavenging test was used to assess the antioxidant activity of the crude extracts. The results of the well and disc diffusion assays showed that Gram-positive bacteria were more sensitive to both extracts compared to Gramnegative bacteria. Meanwhile, the methanolic extract showed higher antibacterial activity on both Gram-positive and Gram-negative bacteria compared to the aqueous extract. The results of the MIC and MBC tests showed that the methanolic extract was bactericidal to both Gram-positive and Gram-negative bacteria. The aqueous extract, however, demonstrated bacteriostatic activity against Gram-negative bacteria and bactericidal activity solely against Gram-positive bacteria. After a 24-h exposure period, a growth reduction assay showed that the methanolic extract could suppress both Gram-positive and Gram-negative bacteria by up to 99%. Meanwhile, the aqueous extract showed an inhibition percentage value ranging from 75% to 96% after an incubation period. The aqueous extract had the lowest antioxidant activity, with an EC50 value of 87.78 µg/mL, whereas the methanolic extract had a fair amount of antioxidant activity when compared to the control (quercetin), according to the DPPH scavenging assay. The present study suggests that C. ternatea extracts as a potential antibacterial agent against pathogenic bacteria with significant antioxidant activity and this activity may be due to the presence of anthocyanin and its derivatives

A review of Malaysian medicinal plants with potential anticancer activity

The global cancer incidence and its high mortality rate indicate limitations in its current treatment and chemotherapeutic strategies. This sparked a worldwide interest in the demand for chemical diversity in searching for therapeutic drugs derived from natural products. Natural products from medicinal plants, whether as pure compounds or crude extracts, offer inexhaustible sources of new drugs because of their unparalleled chemical diversity. This review aims to disseminate detailed information on the anticancer potential of Malaysian medicinal plants, focusing on the bioactive phytochemicals and mechanisms of action against cancer development in both in vitro and in vivo studies. A comprehensive search of PubMed, Google Scholar, Scopus, and ScienceDirect databases was conducted to find relevant articles on the anticancer activity of Malaysian medicinal plants. A total of hundred and twenty-two (122) articles on the anticancer activity of Malaysian medicinal plants was identified and reviewed. Eighty-five (85) plants (in vitro) and 16 plants (in vivo) have been identified to possess anticancer activity. The activity reported was attributed primarily to diverse chemical groups of naturally occurring phytochemicals such as flavonoids, phenolics, glycosides, quercetin, and gallic acid. Henceforth, the findings will hope to aid further research in understanding the underlaying mechanism and the efficiency of the isolation of the bioactive compounds

Oryzias latipes (JAPANESE MEDAKA) as genetic model to study causative genes of epilepsy disease: an in-silico approach

Epilepsy is a chronic neurological disorder that has affected around 50-70 million people worldwide. Various animal models have previously been used in epilepsy research. To expand the knowledge of the disease, a new animal model is suggested to be explored considering the genetic and phenotypic heterogeneity that contributes to the complexity of the disease. This study was undertaken to analyze 14 causative genes of epilepsy disease in Japanese medaka (Oryzias latipes), humans, and the established model of this disease which is zebrafish (Danio rerio) by assessing the variation in the genes by using MEGA X and predicting the functional motif and secondary structure of the proteins by using PROSITE and GORIV respectively. Results from the variation analysis showed the lowest percentage of conserved genes in Japanese medaka was 60%.50% of the genes of Japanese medaka were found to be more conserved than zebrafish in comparison to a human. The functional motifs present in all genes in Japanese medaka showed the same motifs present in humans. All the secondary structures of Japanese medaka genes were predicted to contain the alpha helix, extended strand, and random coil. In conclusion, it can be inferred that Japanese medaka could be a reliable animal model for epilepsy disease

Cytotoxic effect of dillapiole on human breast cancer MCF-7 CELLS

Plant secondary metabolites and their derivatives play a significant role in anticancer drug therapy since they are effective against specific characteristics while reducing side effects. Dillapiole is a phenylpropanoid that holds several bioactivities like anti-fungal, anti-inflammatory, anti-leishmanial, and anti-cancer. This study aims to investigate the possible cytotoxic effect of dillapiole on human breast cancer, MCF-7 cells. Cells were cultured in complete RPMI media and incubated at standard culture conditions. After the cells reached 80% confluency, cells were treated with various concentrations (ranging from 0 μM to 150 μM) of dillapiole and tamoxifen as a positive control. Cells were later incubated at 48 and 72 h. Using WST-1 assay, the cytotoxic effect was determined for both incubation times. Results show tamoxifen inhibited the MCF-7 cells with the IC50 at 75.9 μM and 39.8 μM for 48 and 72 h respectively. Parallel with the positive control results, there was a significant cytotoxic effect of dillapiole against MCF-7 cells at 48- and 72-hr incubation with the IC50 at 92.1 μM and 63.1 μM respectively. Based on these results, dillapiole was cytotoxic against MCF-7 cells and its cytotoxic activity was both in a time and dose-dependent manner (<0.05). The morphological analysis supported the WST-1 assay. Our preliminary finding is in agreement with other previous studies, suggesting that dillapiole appears to be a promising anti-cancer agent and opens a wider possibility of downstream analysis on its underlying cytotoxicity mechanism

Cytotoxic effect of dillapiole on human breast cancer MCF-7 cells

Plant secondary metabolites and their derivatives play a significant role in anticancer drug therapy since they are effective against specific characteristics while reducing side effects. Dillapiole is a phenylpropanoid that holds several bioactivities like anti-fungal, anti-inflammatory, anti-leishmanial, and anti-cancer. This study aims to investigate the possible cytotoxic effect of dillapiole on human breast cancer, MCF-7 cells. Cells were cultured in complete RPMI media and incubated at standard culture conditions. After the cells reached 80% confluency, cells were treated with various concentrations (ranging from 0 μM to 150 μM) of dillapiole and tamoxifen as a positive control. Cells were later incubated at 48 and 72 h. Using WST-1 assay, the cytotoxic effect was determined for both incubation times. Results show tamoxifen inhibited the MCF-7 cells with the IC50 at 75.9 μM and 39.8 μM for 48 and 72 h respectively. Parallel with the positive control results, there was a significant cytotoxic effect of dillapiole against MCF-7 cells at 48- and 72-hr incubation with the IC50 at 92.1 μM and 63.1 μM respectively. Based on these results, dillapiole was cytotoxic against MCF-7 cells and its cytotoxic activity was both in a time and dose-dependent manner (<0.05). The morphological analysis supported the WST-1 assay. Our preliminary finding is in agreement with other previous studies, suggesting that dillapiole appears to be a promising anti-cancer agent and opens a wider possibility of downstream analysis on its underlying cytotoxicity mechanism

Oryzias latipes (Japanese medaka) as genetic model to study causative genes of epilepsy disease : an in-silico approach

Epilepsy is a chronic neurological disorder that has affected around 50-70 million people worldwide. Various animal models have previously been used in epilepsy research. To expand the knowledge of the disease, a new animal model is suggested to be explored considering the genetic and phenotypic heterogeneity that contributes to the complexity of the disease. This study was undertaken to analyze 14 causative genes of epilepsy disease in Japanese medaka (Oryzias latipes), humans, and the established model of this disease which is zebrafish (Danio rerio) by assessing the variation in the genes by using MEGA X and predicting the functional motif and secondary structure of the proteins by using PROSITE and GORIV respectively. Results from the variation analysis showed the lowest percentage of conserved genes in Japanese medaka was 60%.50% of the genes of Japanese medaka were found to be more conserved than zebrafish in comparison to a human. The functional motifs present in all genes in Japanese medaka showed the same motifs present in humans. All the secondary structures of Japanese medaka genes were predicted to contain the alpha helix, extended strand, and random coil. In conclusion, it can be inferred that Japanese medaka could be a reliable animal model for epilepsy disease

Aqueous Extract of Clitoria ternatea Attenuates the Growth of Streptococcus mutans

In the human oral cavity, Streptococcus mutans is often observed and is a major contributor to tooth decay. Increased S. mutans levels may be linked to progressively more severe forms of periodontal disease because root exposure in people with periodontitis increases caries rates. Hence, a new potential antibacterial compound needs to be searched to combat this pathogenic bacterium. The butterfly pea, or Clitoria ternatea is an ornamental plant that has been reported to exhibit antibacterial properties against several bacteria. Thus, the goal of this investigation was to determine how well C. ternatea aqueous (CTA) extract inhibited S. mutans. The disk diffusion assay was performed to access the antibacterial properties of the CTA extract. The efficiency of the extract against the test bacterium was then determined through MIC/MBC determinations and a time-kill study. Meanwhile, the toxicity of the extract was tested using a brine shrimp lethality assay (BSLA). The CTA extract demonstrated substantial antibacterial activity against the test bacterium at a concentration of 200 mg/ml, with a diameter of the inhibition zone of 13.4±0.4 mm, according to the disc diffusion assay. The aqueous extract’s MIC and MBC values were found to be 100 and 400 g/mL, respectively. Time-kill analysis revealed the CTA extract exerted a strong bactericidal effect on S. mutans and this activity was dose-dependent. A scanning electron microscope (SEM) exhibited the bacterial cells experienced severe damage after being exposed to CTA extract including formation cavities, irregular shape, and crumpled cells. Thus, the present study suggested the potential of CTA extract as an antibacterial agent against oral cavity bacteria and can be used in the formulation of natural mouthwash due to no toxicity effect

Assessment of cultivation parameters influencing pectinase production by Aspergillus niger LFP-1 in submerged fermentation

Abstract Background Pectinase is helpful in food and beverage industries, particularly in the preparation of fruit juice, the extraction of vegetable oil, and the fermentation of coffee. The current work aimed to screen Aspergillus niger LFP-1, a recently identified fungal strain, for its ability to produce pectinase and to ascertain the contribution of various physicochemical factors to pectinase production. Results The primary and secondary pectinase activity screenings by Aspergillus niger LFP-1 were performed using pectin screening agar and shake flask system, respectively. The finding revealed that the locally isolated strain is able to secrete favourable pectinase production. Before improvement, the pectinase production was 0.88 ± 0.09 U/mL. However, the improved conditions such as 6 days of the cultivation period, agitation speed of 150 rpm, inoculum size of 1 × 106 spores/mL, 2.5% (w/v) citrus pectin, and 0.4% (w/v) ammonium nitrate could significantly increase pectinase production up to 7.41 ± 0.24 U/mL, representing an 88% increase. In this study, supplementing 2.5% (w/v) citrus pectin to the culture medium as a carbon source increased enzyme production by up to 3.07 ± 0.17 U/mL. Meanwhile, 0.4% (w/v) ammonium nitrate was used as a nitrogen source yielding the highest enzyme activity with a value of 6.86 ± 0.07 U/mL. Conclusion Thus, the locally isolated fungal strain, A. niger LFP-1 has outstanding pectinase-producing capability and can be utilized for the commercial production of pectinase. The improved cultural conditions significantly increase pectinase production and shorten the incubation period from 8 days (before improvement) to 6 days (after improvement)