Cytotoxicity of extract of Malaysian Mitragyna Speciosa Korth and its dominant Alkaloid Mitragynine

Mitragyna speciosa Korth (Kratom), a herb of the Rubiaceae family is indigenous in southeast Asia mainly in Malaysia and Thailand. It is used as an opium substitute and has been increasingly abused by drug addicts in Malaysia. Recently, the potent analgesic effect of plant extract and its dominant alkaloid mitragynine (MIT) were confirmed in vivo and in vitro. MIT acted primarily on μ- and δ-opioid receptors, suggesting that MIT or similar compounds could be promising alternatives for future pain management treatments. However the potential cytotoxicity of this plant is unknown. Therefore, the cytotoxicity of methanol-chloroform extract (MSE) and MIT on human cell lines (HepG2, HEK 293, MCL-5, cHol and SH-SY5Y cells) has been examined. MSE appeared to exhibit dose-dependant inhibition of cell proliferation in all cell lines examined, at concentration > 100 μg/ml with substantial cell death at 1000 μg/ml. SH-SY5Y was the most sensitive cell line examined. MIT showed a similar response. Clonogenicity assay was performed to assess the longer- term effects of MSE and MIT. The colony forming ability of HEK 293 and SH-SY5Y cells was inhibited in a dose-dependant manner. Involvement of metabolism in cytotoxicity was further assessed by clonogenicity assay using rat liver S9 (induced by Arochlor 1254); toxicity increased 10-fold in both cell lines. To determine if cytotoxicity was accompanied by DNA damage, the Mouse lymphoma tk gene mutation assay was used. The results were negative for both MSE and MIT. Studies on the involvement of metabolism in cytotoxicity of MSE and MIT were performed using MCL-5 and it appeared that CYP 2E1 is involved in activation of cytotoxicity. Studies with opioid antagonists were performed using SH-SY5Y cells treated with MSE and MIT. Naloxone (μ and δ receptor antagonists), naltrindole (δ receptor antagonist) and cyprodime hydrobromide (μ receptor antagonist) confirmed that MSE cytotoxicity was associated with μ and δ receptor while MIT mainly acted on μ receptor. Studies on mechanism of MSE and MIT cytotoxicity showed that cell death observed at high dose was preceded by cell cycle arrest, however MSE cell arrest was independent of p53 and p21 while MIT showed opposite result. Studies have been undertaken to examine the nature of this cell death. Morphological examinations showed that cell death induced by MSE was cell type dependant, in which SH-SY5Y cells appeared to die via apoptosis-like cell death while HEK 293 and MCL-5 cells predominantly via necrosis. Biochemical assessments confirmed that MSE induced cell death independent of p53 or caspases pathway while MIT cell death appeared to be associated with p53 and caspases pathway. The involvement of reactive oxygen species (ROS) generation in MSE and MIT mediating cell death was performed using SH-SY5Y cells. The results appeared negative for both MSE and MIT treated cells. Collectively, the findings of these studies suggest that MSE and its dominant alkaloid MIT produced cytotoxicity effects at high dose. Thus, the consumption of Mitragyna speciosa Korth leaves may pose harmful effects to users if taken at high dose and the evidence for involvement of CYP 2E1 in increasing the MSE cytotoxicity suggests that caution may be required if the leaves are to be taken with CYP 2E1 inducers

A Preliminary Study of Genetic Variation of Selected Species of a Lowland Forest at Ayer Hitam Forest Reserve, Selangor

A preliminary study of the extent and pattern of genetic variation of 10 selected species (Shorea parvifolia, Shorea macroptera, Shorea acuminata, Shorea leprosula, Hopea beccariana, Dipterocarpus crinitus, Endospermum malaccensis, Artocarpus elasticus, Palaquium gutta and Macaranga gigantea at Ayer Hitam Forest Reserve was carried out using the horizontal starch gel electrophoresis. Analysis of eight enzymes indicated that they were coded by 9 to 12 loci. Levels of polymorphism and mean of expected heterozygosities of these species ranged from 0.9 to 1.0 and from 0.454 (S. leprosula) to 0.602 (H. beccariana) respectively

Bioconversion of Palm Oil Mill Effluent for the Production of Organic Acids and Polyhydroxyalkanoates

In this study, palm oil mill effluent (POME) was utilized as a substrate for the production of polyhydroxyalkanoate (PHA) by Alcaligenes eutrophus. PHA production was achieved via a two-stage process; organic acids production from POME followed by PHA production from the organic acids. In the first stage, the study focused on optimisation of organic acids production as part of an integrated zero-emission system. Optimisation of organic acids production was performed by batch and continuous anaerobic treatments. In batch treatment, POME was mixed with different inoculum sizes (POME sludge) of 25%, 33%, 50% and 33% diluted sludge (two times) with tapwater in all conical flask. The result showed using inoculum size of 25%, 33% and 50% gave high organic acids production (7.0 g/L) after 4 days/of treatment.Continuous treatment was carried out at four different retention times (2.5, 5, 7.5 and 10 days). POME was mixed with an equal ratio of anaerobic POME sludge in a 12 L stirred-tank bioreactor. The pH was controlled at 7.0 to enhance organic acids production, particularly acetic and propionic. Total organic acids produced were 7.7 to 9.6 g/L for 2.5, 5 and 7.5 days retention times. More than 75% of the chemical oxygen demand (COD) conversion of POME was achieved during anaerobic treatment at 5, 7.5 and 10 days retention times. The best retention time was 5 days which produced 9.53±1.28 g/L of total organic acids, 11700 mg/L COD and 77% COD conversion of POME at steady state. After separation of the organic acids in the treated effluent by ion exchange chromatography, the residual COD of the effluent was less than 100 mg/L which met the effluent discharge standard set for the industry by the Department of Environment of Malaysia. The effluent collected during continuous anaerobic treatment at 5 days retention time was concentrated by evaporation to be used in the second stage for polyhydroxyalkanoate production. The results showed that by evaporation, the organic acids could be concentrated ten-fold to about 100 giL for use as substrates for the fed-batch fermentation. The concentrated organic acids were successfully converted to PHA by Alcaligenes eutrophus strain ATCC 17699 under a non-sterile fermentation system when the initial cell density was kept high at 4 g/L. After 150 hours, 20 g/L cells were obtained with more than 60% (gig) PHA content being produced

Reproductive performance of Kedah-Kelantan cattle at Pusat Ternakan Haiwan Pantai Timur, Malaysia.

Data between 1996 and 2009 at Pusat Ternakan Haiwan Pantai Timur, Malaysia were used to analyse the age at first calving, age at conception, calving interval and mean birth weight of Kedah-Kelantan (KK) cattle. Monthly rainfall from 2006 to 2008 were also analysed to determine the correlation between rainfall and calving rate and calf mortality rate. The results indicated that the mean age at first calving was 1146 ± 148 days, the mean age at conception was 858 ± 148 days, the mean for first, second, and third calving interval were 403 ± 137, 389 ± 116, and 376 ± 90 days, respectively. The result also showed that the calving interval subsequently decreased with increasing parity. No significant differences were found between the three calving intervals (P > 0.05). The mean birth weight was significantly different between female (12.93 kg) and male (13.97 kg) calves (P < 0.05). There was a weak correlation between the calving rate and monthly rainfall pattern (r = 0.26) and between mortality of calves below 3 months of age with rainfall pattern (r = 0.17). The results also showed that there was low negative correlation between mortality rate of calves below 6 months of age (r = -0.28) and total mortality (r = -0.04) with rainfall. In conclusion, Kedah-Kelantan cattle showed very good reproductive performance and are suitable for commercial beef production in Malaysia

The Isolation and Spectroscopic Characterization of Products from the Metatheses of Alkyl Oleates

The products from the metathesis reactions of methyl and ethyl oleate were isolated using column chromatography. The various components obtained were characterized using infrared (IR), 1Hand 13C nuclear magnetic resonance spectroscopy (NMR) and gas chromatography mass spectrometry (GC-MS

Stability and rheology of concentrated of concentrated O/W emulsions based on soybean oil/palm kernel olein blends.

Droplet size distribution and rheological properties of egg yolk-stabilized emulsions were studied before and after storage (25 °C, 30 days). The dispersed phase (70%) of the emulsions was based on soybean oil (SBO) and 10–40% palm kernel olein (PKO) replacements of SBO. Replacement of PKO resulted in a significant increase in droplet mean diameters and a decrease in rheological properties of the emulsions. All emulsion exhibited a gel-like characteristic with storage modulus higher than loss modulus and tan δ greater than 0.3. Significant increase (p < 0.05) was found for droplet mean diameters and rheological properties of the emulsions after storage. Emulsion with fully SBO and the highest PKO replacement (40%) were found to be the most unstable, which was ascribed to a strong flocculation. With 10–30% PKO replacements, the emulsions displayed a better stability after storage, most probably promoted by significant content of short-medium chain fatty acids in PKO

Droplet characterization and stability of soybean oil/palm kernel olein O/W emulsions with the presence of selected polysaccharides.

Droplet characteristics, flow properties and stability of egg yolk-stabilized oil-in-water (O/W) emulsions as affected by the presence of xanthan gum (XG), carboxymethyl cellulose (CMC), guar gum (GG), locust bean gum (LBG) and gum Arabic (AG) were studied. The dispersed phase (40%) of the emulsions was based on soybean oil/palm kernel olein blend (70:30) that partially crystallized during extended storage at 5 °C. In freshly prepared emulsions, the presence of XG, CMC, GG and LBG had significantly decreased the droplet mean diameters. XG, LBG, GG and CMC emulsions exhibited a shear-thinning behavior but AG emulsion exhibited a Bingham plastic behavior and control (without gum) emulsion almost exhibited a Newtonian behavior. Both control and AG emulsions exhibited a severe phase separation after storage (30 days, 5 °C). The microstructure of stored XG emulsion showed the presence of partially coalesced droplets, explaining a large increase in its droplet mean diameters. Increases in droplet mean diameters and decreases in flow properties found for stored GG and LBG emulsions were attributed to droplet coalescence. Nevertheless, the occurrence of droplet coalescence in these emulsions was considered to be small as no free oil could be separated under centrifugation force. Increases in flow properties and excellent stability towards phase separation found for stored CMC emulsion suggested that CMC could retard partial coalescence. Thus, the results support the ability of CMC, GG and LBG in reducing partial coalescence either by providing a sufficiently thick continuous phase or by acting as a protective coating for oil droplets

Monoclonal gammopathy with systemic amylodosis: an evaluation of diagnostic elements

Monoclonal gammopathies result from an overproduction of a single abnormal clone of plasma cell or B lymphocyte that produce an immunologically homogenous immunoglobulin (Ig) commonly referred to as paraprotein or monoclonal (M) protein. The circulating M-protein may consist of an intact immunoglobulin, the light chain only, or (rarely) the heavy chain only. The heavy chain is from one of the five immunoglobulin classes G, A, M, D or E, while the light chain is either kappa (κ) or lambda (λ) in type. Accurate detection and quantitation of monoclonal immunoglobulins is important for the diagnosis and management of monoclonal gammopathies. We report a case of a 71 year old lady with a history of chronic gastritis and recurrent lower respiratory tract infection whereby no specific diagnosis was made until a computed tomography (CT) guided lung biopsy and orogastroduodenoscopy