Optimization of cyclodextrin glycosyltransferase production by response surface methodology approach.

The aim of the study was to optimized cyclodextrin glycosyltransferase (CGTase) production using local isolated strain MK6 which was identified as Bacillus sp. Optimum activity obtained at temperature of 70 °C and the enzyme shows a wide range of pH stability ranging from 4-10 when stored at 4 °C for 24 h and temperature stability ranging from 30-80 °C at 1 h incubation period. The CGTase activity was even maintained at 0.4 U mL-1 at 90 °C for 40 min incubation. Prior to optimization of CGTase production, selection for the best carbon source through detection using modified phenolphthalein method containing different types of starch were performed. Sago starch gave significant result and was used for further optimization using statistical analysis namely Response Surface Methodology (RSM) approach. The optimal calculated values were 3.34% sago starch, initial pH of 10.15 and agitation speed of 187 rpm; with predicted activity of 2.07 U mL-1 of CGTase. These predicted optimal parameters were confirmed in the laboratory and the final CGTase activity obtained was very close to the predicted value of 2.56 U mL-1

Molecular typing of Aeromonas species using RAPD and ERIC-PCR fingerprinting

Two molecular typing methods were used in this study to examine the intra/inter-specific genetic relatedness among the A. hydrophila, A. caviae and A. veronii biovar sobria strains. In the analysis by RAPDPCR and ERIC-PCR, the size for RAPD and ERIC fragments ranged from 0.25 to 10.0 kb with an average number of sixteen and eight bands, respectively. For the inter-species similarity, RAPD could generate 16 major clusters and 36 single isolates at the similarity of 30%. 10 clusters are significant since it grouped the strains according to their own species, however, ERIC-PCR could discriminate the strains into 4 significant clusters and 64 single isolates at the similarity of 50%. Moreover, there was no genetic similarity between food and environmental strains of Aeromonas sp. isolated from different geographical areas as well as from the same geographical area. Eighty five genotypes among the 85 A. hydrophila, A. caviae and A. veronii biovar sobria isolates were generated using RAPD and ERIC-PCR which indicated that the strains were very diverse

Tissue engineering approach to repair abdominal wall defects using cell-seeded bovine tunica vaginalis in a rabbit model

The aim of this study was to engineer skeletal muscle tissue for repair abdominal wall defects. Myoblast were seeded onto the scaffolds and cultivated in vitro for 5 days. Full thickness abdominal wall defects (3 × 4 cm) were created in 18 male New Zealand white rabbits and randomly divided into two equal groups. The defects of the first group were repaired with myoblast-seeded-bovine tunica vaginalis whereas the second group repaired with non-seeded-bovine tunica vaginalis and function as a control. Three animals were sacrificed at 7th, 14th, and 30th days of post-implantation from each group and the explanted specimens were subjected to macroscopic and microscopic analysis. In every case, seeded scaffolds have better deposition of newly formed collagen with neo-vascularisation than control group. Interestingly, multinucleated myotubes and myofibers were only detected in cell-seeded group. This study demonstrated that myoblast-seeded-bovine tunica vaginalis can be used as an effective scaffold to repair severe and large abdominal wall defects with regeneration of skeletal muscle tissue

Angiotensin II stimulates expression of transcription factors c-Jun and c-Fos in cyclosporine induced human gingival fibroblasts

The present study demonstrates that the expression of c-Jun and c-Fos are elevated in gingival fibro- blast cells treated with angiotensin II and cyclosporine. The healthy human gingival tissues were collected and gingival fibroblasts were isolated and cultured. We used RT-PCR and Western blot analysis to identify the expression of c-Jun and c-Fos in cyclosporine and angiotensin II treated human gingival fibroblast cells. We found that angiotensin II in combination with cyclosporine induces c-Jun and c-Fos expressions significantly; however, the angiotensin II antagonist losartan inhibits the expression of c-Jun and c-Fos (p < 0.01). The data suggest that angiotensin II in combination with cyclosporine modulates the expression of c-Jun and c-Fos in human gingival fibroblast cells

Crude ethyl acetate extract of marine microalga, Chaetoceros calcitrans, induces Apoptosis in MDA-MB-231 breast cancer cells

Background: Marine brown diatom Chaetoceros calcitrans and green microalga Nannochloropsis oculata are beneficial materials for various applications in the food, nutraceutical, pharmaceutical and cosmeceutical industries. Objective: This study investigated cytotoxicity of different crude solvent extracts from C. calcitrans and N. oculata against various cancer cell lines. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was carried out to screen the cytotoxic effects of hexane (Hex), dichloromethane (DCM), ethyl acetate, and methanol extract from C. calcitrans and N. oculata toward various cancer cell lines. Flow cytometry cell cycle was used to determine the cell cycle arrest while the mode of cell death was investigated through acridine orange/propidium iodide (AOPI) staining, Annexin V-Fluorescein Isothiocyanate (FITC) and Terminal deoxynucleotidyl transferase-mediated d-UTP Nick End Labeling (TUNEL) assays. Expression profile of apoptotic and proliferative-related genes was then determined using the multiplex gene expression profiler (GeXP). Results: Crude ethyl acetate (CEA) extract of C. calcitrans inhibited growth of MDA-MB-231 cells, with IC 50 of 60 μg/mL after 72 h of treatment. Further studies were conducted to determine the mode of cell death at various concentrations of this extract: 30, 60 and 120 μg/mL. The mode of cell death was mainly apoptosis as shown through apoptosis determination test. The expression data from GeXP showed that caspase-4 was upregulated while B-cell leukemia/lymphoma 2(Bcl-2) was down regulated. Thus, caspase-4 induction endoplasmic reticulum death pathway is believed to be one of the mechanisms underlying the induction of apoptosis while Bcl-2 induced S and G2/M cell cycle phase arrest in MDA-MB-231 cells. Conclusion: CEA extract of C. calcitrans showed the highest cytotoxicity on MDA-MB-231 via apoptosis

Bioactivity studies and chemical constituents of Murraya paniculata (Linn) Jack

Murraya paniculata (Linn) Jack (Orange Jasmine), known as "Kemuning Putih" in Malaysia, has been widely used as food flavor additive in cuisine by local residences. This is due to the strong fragrances of the leaves which make it suitable to be used in Indian and Malay dishes. Besides as a flavoring, leaves, branches, stem barks and roots of the plant are used in folk medicine to treat dysentery and morning sickness. Flowers of the plants are used in cosmetics. Since 1970's, flavonoids and coumarins were isolated from Murraya paniculata, but no further bioactivity has been tested from the isolated compounds. The aim of this paper is to review and update the research related to chemical constituents and bioactivities of Murraya paniculata (L) Jack

Cytotoxic effect of damnacanthal, nordamnacanthal, zerumbone and betulinic acid isolated from Malaysian plant sources

The present study was to evaluate the toxicity of damnacanthal, nordamnacanthal, betulinic acid and zerumbone isolated from local medicinal plants towards leukemia cell lines and immune cells by using MTT assay and flow cytometry cell cycle analysis. The results showed that damnacanthal significantly inhibited HL-60 cells, CEM-SS and WEHI-3B with the IC50 value of 4.0 μg/mL, 8.0 μg/mL and 3.3 μg/mL, respectively. Nordamnacanthal and betulinic acid showed stronger inhibition towards CEM-SS and HL-60 cells with the IC50 value of 5.7 μg/mL and 5.0 μg/mL, respectively. In contrast, Zerumbone was demonstrated to be more toxic towards those leukemia cells with the IC50 value less than 10 μg/mL. Damnacanthal, nordamnacanthal and betulinic acid were not toxic towards 3T3 and PBMC compared to doxorubicin which showed toxicity effects towards 3T3 and PBMC with the IC50 value of 3.0 μg/mL and 28.0 μg/mL, respectively. The cell cycle analysis exhibited that damnacanthal exerted its toxicity effect towards HL-60 cells by inducing apoptosis with value of 25% after 72 hours treatment. Thus, these compounds could be the potential anticancer drug with less toxic side effect

Estrogenic activity of Elaeis guineensis leaf

To assess the estrogenic activities of Oil Palm Fronds (Elaeis guineensis) extracts a vaginal cytology assay in normal and ovariectomized rats were used. The alcoholic extract of Elaeis guineensis (OPLME) was administered orally to adult normal cycling and ovareiectomized Sprague/Dawley rats. Bilaterally ovariectomized rats were divided into four groups (n = 8) receiving different treatments, consisting of vehicle, 150 and 300 mg/kg body weight OPLME and Premarin (conjugated estrogens) at a dose of 2.5 mg/kg body weight. Estrogenic activity was assessed by taking percentage vaginal cornification and uterine wet weight as parameters of assessment. The OPLME resulted in an irregular estrous cycle with lengthened estrus in dose dependent manner. Restoration of normal estrous cycles after withdrawal of treatment indicates the reversible effect of alcoholic extract in normal rats. OPLME administration produced statistically significant (P < 0.001), 2.54/fold increase in circulating 17 β /estradiol levels. OPLME showed a significant increase in percentage vaginal cornification, and uterine wet weight (P < 0.001), compared to the control in a dose dependent manner in ovariectomized rat. This estrogenic property of OPLME may be a possible explanation for the OPLME effects on the blood lipid profile. The estrogenic activity shown by OPLME can be attributed to the presence of flavonoids and phenolic compounds

Anti-proliferative effects of pandan leaves (Pandanus amarylfolius), kantan flower(Etlingera elatior) and turmeric leaves (Curcuma longa).

Purpose – The purpose of this paper is to screen cytotoxic activities of commonly used culinary plants in Malaysia, Pandanus amaryllifolius (daun pandan), Curcuma longa (turmeric leaves) and Etlingera elatior (kantan flower) against selected cancer cell lines. Design/methodology/approach – Plant samples were extracted exhaustively with ethanol and concentrated under rotary evaporator. Cytotoxic evaluation was carried out with plant extracts(0-100mg/ml) using 72-h MTT assay. Findings – Exposure of plant extracts reduced cell viability of HepG2 (hepatocellular carcinoma),HT-29 (colon carcinoma), MDA-MB-231 (non-hormone-dependent breast cancer), MCF-7(hormone-dependent breast cancer) and HeLa (cervical cancer); 50 percent inhibitory values (IC50) were obtained for MDA-MB-231, HepG2, HT-29. Extracts within the concentrations of 10-100mg/ml were found not to be effective against proliferation of MCF-7 and HeLa