A New Approach In Protein Folding Studies Revealed The Potential Site For Nucleation Center

A new approach to predict the 3D structures of proteins by combining the knowledge-based method and Molecular Dynamics Simulation is presented on the chicken villin headpiece subdomain (HP-36). Comparative modeling is employed as the knowledge-based method to predict the core region (Ala9-Asn28) of the protein while the remaining residues are built as extended regions (Met1-Lys8; Leu29-Phe36) which then further refined using Molecular Dynamics Simulation for 120 ns. Since the core region is built based on a high sequence identity to the template (65%) resulting in RMSD of 1.39 Å from the native, it is believed that this well-developed core region can act as a 'nucleation center' for subsequent rapid downhill folding. Results also demonstrate that the formation of the non-native contact which tends to hamper folding rate can be avoided. The best 3D model that exhibits most of the native characteristics is identified using clustering method which then further ranked based on the conformational free energies. It is found that the backbone RMSD of the best model compared to the NMR-MDavg is 1.01 Å and 3.53 Å, for the core region and the complete protein, respectively. In addition to this, the conformational free energy of the best model is lower by 5.85 kcal/mol as compared to the NMR-MDavg. This structure prediction protocol is shown to be effective in predicting the 3D structure of small globular protein with a considerable accuracy in much shorter time compared to the conventional Molecular Dynamics simulation alone.

The Effect of Pleurotus sajor-caju (PSC) Addition on the Nutritional Composition and Sensory Properties of Poultry-Based Patty

The nutrient composition and sensory properties of poultry-based patties (PBPs) incorporated with various levels of grey oyster mushroom (Pleurotus sajor-caju, PSC) were studied. The PBPs were formulated with either 0%, 25% or 50% of fresh ground PSC. Results show poultry patty formulated with 25% PSC had protein content of 17.46% lower than the control patty which had 18.13% but it was not significant. Meanwhile, both cooked poultry patties containing 25% and 50% PSC significantly recorded lower concentration of fat at 10.67% and 7.15%, respectively. On the other hand, poultry patty added with 50% ground PSC shows the highest concentration of total dietary fibre (TDF) of 4.90 g/100g compared to poultry patty containing 25% of mushroom (3.40 g/100g) and to the control (1.90g/100g). In addition, patty incorporated with 25% PSC had moisture content of 57.91% which is significantly lower than patty formulated with 50% which had moisture of 61.80%. In the sensory evaluation, there were no differences recorded in all sensory attributes of PSC-based patties judged by untrained panelists. In conclusion, the addition of PSC to replace poultry meat can be recommended for the purpose of lowering production cost, enhancing nutritional composition and maintaining the acceptability of poultry patties.

Yield, Yield Components, Soil Minerals and Aroma of KDML 105 Rice in Tungkularonghai, Roi-Et,Thailand

Pearson-s correlation coefficient and sequential path analysis has been used for determining the interrelationship among yield, yield components, soil minerals and aroma of Khao Dawk Mali (KDML) 105 rice grown in the area of Tungkularonghai in Roi-Et province, located in the northeast of Thailand. Pearson-s correlation coefficient in this study showed that the number of panicles was the only factor that had positive significant (0.790**) effect on grain yield. Sequential path analysis revealed that the number of panicles followed by the number of fertile spikelets and 100-grain weight were the first-order factors which had positive direct effects on grain yield. Whereas, other factors analyzed had indirect effects influencing grain yield. This study also indicated that no significant relationship was found between the aroma level and any of the factors analyzed.

Plaque Formation of Toxoplasma gondii in Vero Cells using Carboxymethylcellulose

Toxoplasma gondii is an intracellular parasite capable of infecting all nucleated cells in a diverse array of species. Toxoplasma plaque assay have been described using Bacto Agar. Because of its experimental advantages carboxymethyl cellulose overlay, medium viscosity was choosing and the aim of this work was to develop alternative method for formation of T. gondii plaques. Tachyzoites were inoculated onto monolayers of Vero cells and cultured at 37° C under 5 % CO2. The cultures were followed up by microscopy inspection. Small plaques were visible by naphtol blue stain 4 days after infection. Larger plaques could be observed by day 10 of culture. The carboxymethyl cellulose is a cheap reagent and the methodology is easier, faster than assays under agar overlay. This is the first description of the carboxymethyl cellulose overlay use for obtaining the formation of T. gondii plaques and may be useful in consequent obtaining tachyzoites for detailed studies.

Microbial Oil Production by Isolated Oleaginous Yeast Torulaspora globosa YU5/2

Microbial oil was produced by soil isolated oleaginous yeast YU5/2 in flask-batch fermentation. The yeast was identified by molecular genetics technique based on sequence analysis of the variable D1/D2 domain of the large subunit (26S) ribosomal DNA and it was identified as Torulaspora globosa. T. globosa YU5/2 supported maximum values of 0.520 g/L/d, 0.472 g lipid/g cells, 4.16 g/L, and 0.156 g/L/d for volumetric lipid production rate, and specific yield of lipid, lipid concentration, and specific rate of lipid production respectively, when culture was performed in nitrogen-limiting medium supplemented with 80g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sweet potato tubers hydrolysates as carbon source.

Interactions between Cells and Nanoscale Surfaces of Oxidized Silicon Substrates

The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

Detection of Oxidative Stress Induced by Mobile Phone Radiation in Tissues of Mice using 8-Oxo-7, 8-Dihydro-2'-Deoxyguanosine as a Biomarker

We investigated oxidative DNA damage caused by radio frequency radiation using 8-oxo-7, 8-dihydro-2'- deoxyguanosine (8-oxodG) generated in mice tissues after exposure to 900 MHz mobile phone radio frequency in three independent experiments. The RF was generated by a Global System for Mobile Communication (GSM) signal generator. The radio frequency field was adjusted to 25 V/m. The whole body specific absorption rate (SAR) was 1.0 W/kg. Animals were exposed to this field for 30 min daily for 30 days. 24 h post-exposure, blood serum, brain and spleen were removed and DNA was isolated. Enzyme-linked immunosorbent assay (ELISA) was used to measure 8-oxodG concentration. All animals survived the whole experimental period. The body weight of animals did not change significantly at the end of the experiment. No statistically significant differences observed in the levels of oxidative stress. Our results are not in favor of the hypothesis that 900 MHz RF induces oxidative damage.

Construction of Recombinant E.coli Expressing Fusion Protein to Produce 1,3-Propanediol

In this study, a synthetic pathway was created by assembling genes from Clostridium butyricum and Escherichia coli in different combinations. Among the genes were dhaB1 and dhaB2 from C. butyricum VPI1718 coding for glycerol dehydratase (GDHt) and its activator (GDHtAc), respectively, involved in the conversion of glycerol to 3-hydroxypropionaldehyde (3-HPA). The yqhD gene from E.coli BL21 was also included which codes for an NADPHdependent 1,3-propanediol oxidoreductase isoenzyme (PDORI) reducing 3-HPA to 1,3-propanediol (1,3-PD). Molecular modeling analysis indicated that the conformation of fusion protein of YQHD and DHAB1 was favorable for direct molecular channeling of the intermediate 3-HPA. According to the simulation results, the yqhD and dhaB1 gene were assembled in the upstream of dhaB2 to express a fusion protein, yielding the recombinant strain E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP41Y3). Strain BP41Y3 gave 10-fold higher 1,3-PD concentration than E. coliBL21 (DE3)//pET22b+::yqhD-dhaB1_dhaB2 (strain BP31Y2) expressing the recombinant enzymes simultaneously but in a non-fusion mode. This is the first report using a gene fusion approach to enhance the biological conversion of glycerol to the value added compound 1,3- PD.

Salicylhydroxamic Acid Inhibits the Growth of Candida albicans

Candida spp. are common and aggressive pathogens. Because of the growing resistance of Candida spp. to current antifungals, novel targets, found in Candida spp. but not in humans or other flora, have to be identified. The alternative oxidase (AOX) is one such possibility. This enzyme is insensitive to cyanide, but is sensitive to compounds such as salicylhydroxamic acid (SHAM), disulfiram and n-alkyl gallates. The growth Candida albicans was inhibited by SHAM (Ki = 9-15 mM) and cyanide (Ki = 2-4 mM), albeit to differing extents. The rate of O2 uptake was inhibited by less than 10% by 25 mM SHAM and by about 90% by 250 μM KCN. Although SHAM substantially inhibited the growth of C. albicans, it is unlikely that the inhibition of AOX was the cause. Salicylhydroxamic acid is used therapeutically in the treatment of urinary tract infections and urolithiasis, but it also has some potential in the treatment of C. albicans infection.

Adsorption Studies on the Removal of Pesticides(Carbofuran) using Activated Carbon from Rice Straw Agricultural Waste

In this study, we used a two-stage process and potassium hydroxide (KOH) to transform waste biomass (rice straw) into activated carbon and then evaluated the adsorption capacity of the waste for removing carbofuran from an aqueous solution. Activated carbon was fast and effective for the removal of carbofuran because of its high surface area. The native and carbofuran-loaded adsorbents were characterized by elemental analysis. Different adsorption parameters, such as the initial carbofuran concentration, contact time, temperature and pH for carbofuran adsorption, were studied using a batch system. This study demonstrates that rice straw can be very effective in the adsorption of carbofuran from bodies of water.

Neutralization of Alkaline Waste-Waters using a Blend of Microorganisms

The efficient operation of any biological treatment process requires pre-treatment of incompatible pollutants such as acids, bases, oil, toxic substances, etc. which hamper the treatment of other major components which are otherwise degradable. The pre-treatment of alkaline waste-waters, generated from various industries like textile, paper & pulp, potato-processing industries, etc., having a pH of 10 or higher, is essential. The pre-treatment, i.e., neutralization of such alkaline waste-waters can be achieved by chemical as well as biological means. However, the biological pretreatment offers better package over the chemical means by being safe and economical. The biological pre-treatment can be accomplished by using a blend of microorganisms able to withstand such harsh alkaline conditions. In the present study, for the proper pre-treatment of alkaline waste-waters, a package of alkalophilic bacteria is formulated to neutralise the alkaline pH of the industrial waste-waters. The developed microbial package is cost-effective as well as environmental friendly.

Microalgal Lipid Production by Microalgae Chlorella sp. KKU-S2

The objective of this work is to produce heterotrophic microalgal lipid in flask-batch fermentation. Chlorella sp. KKU-S2 supported maximum values of 0.374 g/L/d, 0.478 g lipid/g cells, and 0.112 g/L/d for volumetric lipid production rate, and specific yield of lipid, and specific rate of lipid production, respectively when culture was performed on BG-11 medium supplemented with 50g/L glucose. Among the carbon sources tested, maximum cell yield coefficient (YX/S, g/L), maximum specific yield of lipid (YP/X, g lipid/g cells) and volumetric lipid production rate (QP, g/L/d) were found of 0.728, 0.237, and 0.619, respectively, using sugarcane molasses as carbon source. The main components of fatty acid from extracted lipid were palmitic acid, stearic acid, oleic acid and linoleic acid which similar to vegetable oils and suitable for biodiesel production.

Reduce of Fermentation Time in Composting Process by Using a Special Microbial Consortium

Composting is the process in which municipal solid waste (MSW) and other organic waste materials such as biosolids and manures are decomposed through the action of bacteria and other microorganisms into a stable granular material which, applied to land, as soil conditioner. Microorganisms, especially those that are able to degrade polymeric organic material have a key role in speed up this process. The aim of this study has been established to isolation of microorganisms with high ability to production extracellular enzymes for degradation of natural polymers that are exists in MSW for decreasing time of degradation phase. Our experimental study for isolation designed in two phases: in first phase we isolated degrading microorganism with selected media that consist a special natural polymer such as cellulose, starch, lipids and etc as sole source of carbon. In second phase we selected microorganism that had high degrading enzyme production with enzymatic assay for seed production. However, our findings in pilot scale have indicated that usage of this microbial consortium had high efficiency for decreasing degradation phase.

A Model to Study the Effect of Excess Buffers and Na+ Ions on Ca2+ Diffusion in Neuron Cell

Calcium is a vital second messenger used in signal transduction. Calcium controls secretion, cell movement, muscular contraction, cell differentiation, ciliary beating and so on. Two theories have been used to simplify the system of reaction-diffusion equations of calcium into a single equation. One is excess buffer approximation (EBA) which assumes that mobile buffer is present in excess and cannot be saturated. The other is rapid buffer approximation (RBA), which assumes that calcium binding to buffer is rapid compared to calcium diffusion rate. In the present work, attempt has been made to develop a model for calcium diffusion under excess buffer approximation in neuron cells. This model incorporates the effect of [Na+] influx on [Ca2+] diffusion,variable calcium and sodium sources, sodium-calcium exchange protein, Sarcolemmal Calcium ATPase pump, sodium and calcium channels. The proposed mathematical model leads to a system of partial differential equations which have been solved numerically using Forward Time Centered Space (FTCS) approach. The numerical results have been used to study the relationships among different types of parameters such as buffer concentration, association rate, calcium permeability.