Quantitative Determination of Free Radical Scavenging Activity and Anti-tumor Activity of Some Myanmar Herbal Plants

Antioxidant activities of ethanolic extracts of Ardisia japonica Blume., Ageartum conyzoides Linn., and Cocculus hirsutus Linn Diels. leaves was determined qualitatively and quantitatively in this research. 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical solution was used to investigate free radical scavenging activity of these leaves extracts. Ascorbic acid (Vitamin C) was used as the standard. In the present investigation, it is found that all of these extracts have remarkable antioxidant activities. The EC50 values of these ethanolic extracts were 12.72 μg/ml for A. japonica, 15.19 μg/ml for A. conyzoides, 10.68 μg/ml for C. hirsutus respectively. Among these Myanmar medicinal plants, C. hirsutus showed higher antioxidant activities as well as free radical scavenging activity than black tea (Camellia sinensis), the famous antioxidant, and A. japonica and A. conyzoides showed a rather lower antioxidant activity than tea extracts. According to results from bioassay with carrot discs infected with Agrobacterium tumefaciens, all extracts showed anti-tumor activity after 3 weeks of incubation. No gall was detected in carrot disks treated with C. hirsutus and A. japonica extracts in the dose of 100ppm and in carrot discs treated with A. conyzoides extract in the dose of 1000 ppm. Therefore, the research clearly indicates that these weedy plants of dry farm land are exceptionally advantageous for human health.

Effect of the Seasonal Variation in the Extrinsic Incubation Period on the Long Term Behavior of the Dengue Hemorrhagic Fever Epidemic

The incidences of dengue hemorrhagic disease (DHF) over the long term exhibit a seasonal behavior. It has been hypothesized that these behaviors are due to the seasonal climate changes which in turn induce a seasonal variation in the incubation period of the virus while it is developing the mosquito. The standard dynamic analysis is applied for analysis the Susceptible-Exposed- Infectious-Recovered (SEIR) model which includes an annual variation in the length of the extrinsic incubation period (EIP). The presence of both asymptomatic and symptomatic infections is allowed in the present model. We found that dynamic behavior of the endemic state changes as the influence of the seasonal variation of the EIP becomes stronger. As the influence is further increased, the trajectory exhibits sustained oscillations when it leaves the chaotic region.

Enzymatic Saccharification of Dilute Alkaline Pre-treated Microalgal (Tetraselmis suecica) Biomass for Biobutanol Production

Enzymatic saccharification of biomass for reducing sugar production is one of the crucial processes in biofuel production through biochemical conversion. In this study, enzymatic saccharification of dilute potassium hydroxide (KOH) pre-treated Tetraselmis suecica biomass was carried out by using cellulase enzyme obtained from Trichoderma longibrachiatum. Initially, the pre-treatment conditions were optimised by changing alkali reagent concentration, retention time for reaction, and temperature. The T. suecica biomass after pre-treatment was also characterized using Fourier Transform Infrared Spectra and Scanning Electron Microscope. These analyses revealed that the functional group such as acetyl and hydroxyl groups, structure and surface of T. suecica biomass were changed through pre-treatment, which is favourable for enzymatic saccharification process. Comparison of enzymatic saccharification of untreated and pre-treated microalgal biomass indicated that higher level of reducing sugar can be obtained from pre-treated T. suecica. Enzymatic saccharification of pre-treated T. suecica biomass was optimised by changing temperature, pH, and enzyme concentration to solid ratio ([E]/[S]). Highest conversion of carbohydrate into reducing sugar of 95% amounted to reducing sugar yield of 20 (wt%) from pre-treated T. suecica was obtained from saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1 after 72 h of incubation. Hydrolysate obtained from enzymatic saccharification of pretreated T. suecica biomass was further fermented into biobutanol using Clostridium saccharoperbutyliticum as biocatalyst. The results from this study demonstrate a positive prospect of application of dilute alkaline pre-treatment to enhance enzymatic saccharification and biobutanol production from microalgal biomass.

Solid State Fermentation of Cassava Peel with Trichoderma viride (ATCC 36316) for Protein Enrichment

Solid state fermentation of cassava peel with emphasis on protein enrichment using Trichoderma viride was evaluated. The effect of five variables: moisture content, pH, particle size (p), nitrogen source and incubation temperature; on the true protein and total sugars of cassava peel was investigated. The optimum fermentation period was established to be 8 days. Total sugars were 5-fold higher at pH 6 relative to pH 4 and 7-fold higher when cassava peels were fermented at 30oC relative to 25oC as well as using ammonium sulfate as the nitrogen source relative to urea or a combination of both. Total sugars ranged between 123.21mg/g at 50% initial moisture content to 374mg/g at 60% and from 190.59mg/g with particle size range of 2.00>p>1.41mm to 310.10mg/g with 4.00>p>3.35mm.True protein ranged from 229.70 mg/g at pH 4 to 284.05 mg/g at pH 6; from 200.87 mg/g with urea as nitrogen source and to 254.50mg/g with ammonium sulfate; from 213.82mg/g at 50% initial moisture content to 254.50mg/g at 60% moisture content, from 205.75mg/g in cassava peel with 5.6>p> 4.75mm to 268.30 in cassava peel with particle size 4.00>p>3.35mm, from 207.57mg/g at 25oC to 254.50mg/g at 30oC Cassava peel with particle size 4.00>p>3.35 mm and initial moisture content of 60% at pH 6.0, 30oC incubation temperature with ammonium sulfate (10g N / kg substrate) was most suitable for protein enrichment with Trichoderma viride. Crude protein increased from 4.21 % in unfermented cassava peel samples to 10.43 % in fermented samples.

Growth Behaviors, Thermostable Direct Hemolysin Secretion and Fatty Acid Profiles of Acid-adapted and Non-adapted Vibrio parahaemolyticus

Three strains of Vibrio parahaemolyticus (690, BCRC 13023 and BCRC 13025) implicated in food poisoning outbreaks in Taiwan were subjected to acid adaptation at pH 5.5 for 90 min. The growth behaviors of acid-adapted and non-adapted V. parahaemolyticus in the media supplemented with various nitrogen and carbon sources were investigated. The effects of acid adaptation on the thermostable direct hemolysin (TDH) secretion and fatty acid profiles of V. parahaemolyticus were also examined. Results showed that acid-adapted and non-adapted V. parahaemolyticus 690, BCRC 13023 and BCRC 13025 grew similarly in TSB-3% NaCl and basal media supplemented with various carbon and nitrogen sources during incubation period. Higher TDH secretion was noted with V. parahaemolyticus 690 among the three strains. However, acid-adapted strains produced less amounts of TDH than non-adapted strains when they were grown in TSB-3% NaCl. Additionally, acid adaptation increased the ratio of SFA/USFA in cells of V. parahaemolyticus strains.

Leatherback Turtle (Dermochelys coriacea) after Incubation Eggshell in Andaman Sea, Thailand Study: Microanalysis on Ultrastructure and Elemental Composition

There are few studies on eggshell of leatherback turtle which is endangered species in Thailand. This study was focusing on the ultrastructure and elemental composition of leatherback turtle eggshells collected from Andaman Sea Shore, Thailand during the nesting season using scanning electron microscope (SEM). Three eggshell layers of leatherback turtle; the outer cuticle layer or calcareous layer, the middle layer or middle multistrata layer and the inner fibrous layer were recognized. The outer calcareous layer was thick and porosity which consisted of loose nodular units of various crystal shapes and sizes. The loose attachment between these units resulted in numerous spaces and openings. The middle layer was compact thick with several multistrata and contained numerous openings connecting to both outer cuticle layer and inner fibrous layer. The inner fibrous layer was compact and thin, and composed of numerous reticular fibers. Energy dispersive X-ray microanalysis detector revealed energy spectrum of X-rays character emitted from all elements on each layer. The percentages of all elements were found in the following order: carbon (C) > oxygen (O) > calcium (Ca) > sulfur (S) > potassium (K) > aluminum (Al) > iodine (I) > silicon (Si) > chlorine (Cl) > sodium (Na) > fluorine (F) > phosphorus (P) > magnesium (Mg). Each layer consisted of high percentage of CaCO3 (approximately 98%) implying that it was essential for turtle embryonic development. A significant difference was found in the percentages of Ca and Mo in the 3layers. Moreover, transition metal, metal and toxic non-metal contaminations were found in leatherback turtle eggshell samples. These were palladium (Pd), molybdenum (Mo), copper (Cu), aluminum (Al), lead (Pb), and bromine (Br). The contamination elements were seen in the outer layers except for Mo. All elements were readily observed and mapped using Smiling program. X-ray images which mapped the location of all elements were showed. Calcium containing in the eggshell appeared in high contents and was widely distributing in clusters of the outer cuticle layer to form CaCO3 structure. Moreover, the accumulation of Na and Cl was observed to form NaCl which was widely distributing in 3 eggshell layers. The results from this study would be valuable on assessing the emergent success in this endangered species.

Cloning and Over Expression of an Aspergillus niger XP Phytase Gene (phyA) in Pichia pastoris

A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.

Pollution Induced Community Tolerance(PICT) of Microorganisms in Soil Incubated with Different Levels of PB

Soil microbial activity is adversely affected by pollutants such as heavy metals, antibiotics and pesticides. Organic amendments including sewage sludge, municipal compost and vermicompost are recently used to improve soil structure and fertility. But, these materials contain heavy metals including Pb, Cd, Zn, Ni and Cu that are toxic to soil microorganisms and may lead to occurrence of more tolerant microbes. Among these, Pb is the most abundant and has more negative effect on soil microbial ecology. In this study, Pb levels of 0, 100, 200, 300, 400 and 500 mg Pb [as Pb(NO3)2] per kg soil were added to the pots containing 2 kg of a loamy soil and incubated for 6 months at 25°C with soil moisture of - 0.3 MPa. Dehydrogenase activity of soil as a measure of microbial activity was determined on 15, 30, 90 and 180 days after incubation. Triphenyl tetrazolium chloride (TTC) was used as an electron acceptor in this assay. PICTs (€IC50 values) were calculated for each Pb level and incubation time. Soil microbial activity was decreased by increasing Pb level during 30 days of incubation but the induced tolerance appeared on day 90 and thereafter. During 90 to 180 days of incubation, the PICT was gradually developed by increasing Pb level up to 200 mg kg-1, but the rate of enhancement was steeper at higher concentrations.

Study on the Effect of Sulphur, Glucose, Nitrogen and Plant Residues on the Immobilization of Sulphate-S in Soil

In order to evaluate the relationship between the sulphur (S), glucose (G), nitrogen (N) and plant residues (st), sulphur immobilization and microbial transformation were monitored in five soil samples from 0-30 cm of Bastam farmers fields of Shahrood area following 11 treatments with different levels of Sulphur (S), glucose (G), N and plant residues (wheat straw) in a randomized block design with three replications and incubated over 20, 45 and 60 days, the immobilization of SO4 -2-S presented as a percentage of that added, was inversely related to its addition rate. Additions of glucose and plant residues increased with the C-to-S ratio of the added amendments, irrespective of their origins (glucose and plant residues). In the presence of C sources (glucose or plant residues). N significantly increased the immobilization of SO4 -2-S, whilst the effect of N was insignificant in the absence of a C amendment. In first few days the amounts of added SO4 -2-S immobilized were linearly correlated with the amounts of added S recovered in the soil microbial biomass. With further incubation the proportions of immobilized SO4 -2-S remaining as biomass-S decreased. Decrease in biomass-S was thought to be due to the conversion of biomass-S into soil organic-S. Glucose addition increased the immobilization (microbial utilization and incorporation into the soil organic matter) of native soil SO4 -2-S. However, N addition enhance the mineralization of soil organic-S, increasing the concentration of SO4 - 2-S in soil.

Temperature-dependent Structural Perturbation of Tuna Myoglobin

To unveil the mechanism of fast autooxidation of fish myoglobins, the effect of temperature on the structural change of tuna myoglobin was investigated. Purified myoglobin was subjected to preincubation at 5, 20, 50 and 40oC. Overall helical structural decay through thermal treatment up to 95oC was monitored by circular dichroism spectrometry, while the structural changes around the heme pocket was measured by ultraviolet/visible absorption spectrophotometry. As a result, no essential structural change of myoglobin was observed under 30oC, roughly equivalent to their body temperature, but the structure was clearly damaged at 40oC. The Soret band absorption hardly differed irrespective of preincubation temperature, suggesting that the structure around the heme pocket was not perturbed even after thermal treatment.

Isolation and Identification of an Acetobacter Strain from Iranian White-Red Cherry with High Acetic Acid Productivity as a Potential Strain for Cherry Vinegar Production in Foodand Agriculture Biotechnology

According to FDA (Food and Drug Administration of the United States), vinegar is definedas a sour liquid containing at least 4 grams acetic acid in 100 cubic centimeter (4% solution of acetic acid) of solution that is produced from sugary materials by alcoholic fermentation. In the base of microbial starters, vinegars could be contained of more than 50 types of volatile and aromatic substances that responsible for their sweet taste and smelling. Recently the vinegar industry has a great proportion in agriculture, food and microbial biotechnology. The acetic acid bacteria are from the family Acetobacteraceae. Regarding to the latest version of Bergy-s Mannual of Systematic Bacteriology that has categorized bacteria in the base of their 16s RNA differences, the most important acetic acid genera are included Acetobacter (genus I), Gluconacetobacter (genus VIII) and Gluconobacter (genus IX). The genus Acetobacter that is primarily used in vinegar manufacturing plants is a gram negative, obligate aerobe coccus or rod shaped bacterium with the size 0.6 - 0.8 X 1.0 - 4.0 μm, nonmotile or motile with peritrichous flagella and catalase positive – oxidase negative biochemically. Some strains are overoxidizer that could convert acetic acid to carbon dioxide and water.In this research one Acetobacter native strain with high acetic acid productivity was isolated from Iranian white – red cherry. We used two specific culture media include Carr medium [yeast extract, 3%; ethanol, 2% (v/v); bromocresol green, 0.002%; agar, 2% and distilled water, 1000 ml], Frateur medium [yeast extract, 10 g/l; CaCO3, 20 g/l; ethanol, 20 g/l; agar, 20 g/l and distilled water, 1000 ml] and an industrial culture medium. In addition to high acetic acid production and high growth rate, this strain had a good tolerance against ethanol concentration that was examined using modified Carr media with 5%, 7% and 9% ethanol concentrations. While the industrial strains of acetic acid bacteria grow in the thermal range of 28 – 30 °C, this strain was adapted for growth in 34 – 36 °C after 96 hours incubation period. These dramatic characteristics suggest a potential biotechnological strain in production of cherry vinegar with a sweet smell and different nutritional properties in comparison to recent vinegar types. The lack of growth after 24, 48 and 72 hours incubation at 34 – 36 °C and the growth after 96 hours indicates a good and fast thermal flexibility of this strain as a significant characteristic of biotechnological and industrial strains.

The Kinetic of Biodegradation Lignin in Water Hyacinth (Eichhornia Crassipes) by Phanerochaete Chrysosporium using Solid State Fermentation (SSF) Method for Bioethanol Production, Indonesia

Lignocellulosic materials are considered the most abundant renewable resource available for the Bioethanol Production. Water Hyacinth is one of potential raw material of the world-s worst aquatic plant as a feedstock to produce Bioethanol. The purposed this research is obtain reduced of matter for biodegradation lignin in Biological pretreatment with White Rot Fungi eg. Phanerochaete Chrysosporium using Solid state Fermentation methods. Phanerochaete Chrysosporium is known to have the best ability to degraded lignin, but simultaneously it can also degraded cellulose and hemicelulose. During 8 weeks incubation, water hyacinth occurred loss of weight reached 34,67%, while loss of lignin reached 67,21%, loss of cellulose reached 11,01% and loss of hemicellulose reached 36,56%. The kinetic of losses lignin using regression linear plot, the results is obtained constant rate (k) of reduction lignin is -0.1053 and the equation of reduction of lignin is y = wo - 0, 1.53 x

Mathematical Model of Dengue Disease with the Incubation Period of Virus

Dengue virus is transmitted from person to person through the biting of infected Aedes Aegypti mosquitoes. DEN-1, DEN-2, DEN-3 and DEN-4 are four serotypes of this virus. Infection with one of these four serotypes apparently produces permanent immunity to it, but only temporary cross immunity to the others. The length of time during incubation of dengue virus in human and mosquito are considered in this study. The dengue patients are classified into infected and infectious classes. The infectious human can transmit dengue virus to susceptible mosquitoes but infected human can not. The transmission model of this disease is formulated. The human population is divided into susceptible, infected, infectious and recovered classes. The mosquito population is separated into susceptible, infected and infectious classes. Only infectious mosquitoes can transmit dengue virus to the susceptible human. We analyze this model by using dynamical analysis method. The threshold condition is discussed to reduce the outbreak of this disease.

Middle East towards Incubator Benefits: Case Studies

In the context of business incubation (BI) as strategic enablers, this paper critically reviews the literature relating to the strategic benefits of BI in the Middle East. The taxonomy of BI benefits in the strategic elements on 1) type, 2) financial model, 3) services, 4) objectives, 5) number of clients, 6) number of graduates, and 7) jobs creation. Understanding the importance of BI benefits can be significant in the economic development although most incubators lead to diversify the economy. Thus, taxonomies of the benefits of BI are produced from both the academic literature and published case studies. In this way, a classification of strategic benefits elements as they relate to incubators has been developed to provide a greater understanding of the benefits needed to obtain a specific element. The result of this paper is Business incubators is aimed entrepreneurship, jobs creation, research commercialization and profitable enterprises in Middle Eastern countries.

Production of Cellulases by Aspergillus Heteromorphus from Wheat Straw under Submerged Fermentation

To investigate the production of cellulases from Aspergillus heteromorphus, submerged fermentation was performed using wheat straw as substrate. Optimization of saccharification conditions like pH, temperature and time were studied. Highest reducing sugar was released on 5th day at 5 pH, 30° C temperature. When A. heteromorphous was grown on wheat straw in submerged fermentation after 5 days incubation at 30 ° C, 3.2 IU/ml and 83 IU/ml, filter paper activity and CMCase activity respectively.

Optimization of Ethanol Fermentation from Pineapple Peel Extract Using Response Surface Methodology (RSM)

Ethanol has been known for a long time, being perhaps the oldest product obtained through traditional biotechnology fermentation. Agriculture waste as substrate in fermentation is vastly discussed as alternative to replace edible food and utilization of organic material. Pineapple peel, highly potential source as substrate is a by-product of the pineapple processing industry. Bio-ethanol from pineapple (Ananas comosus) peel extract was carried out by controlling fermentation without any treatment. Saccharomyces ellipsoides was used as inoculum in this fermentation process as it is naturally found at the pineapple skin. In this study, the capability of Response Surface Methodology (RSM) for optimization of ethanol production from pineapple peel extract using Saccharomyces ellipsoideus in batch fermentation process was investigated. Effect of five test variables in a defined range of inoculum concentration 6- 14% (v/v), pH (4.0-6.0), sugar concentration (14-22°Brix), temperature (24-32°C) and time of incubation (30-54 hrs) on the ethanol production were evaluated. Data obtained from experiment were analyzed with RSM of MINITAB Software (Version 15) whereby optimum ethanol concentration of 8.637% (v/v) was determined. The optimum condition of 14% (v/v) inoculum concentration, pH 6, 22°Brix, 26°C and 30hours of incubation. The significant regression equation or model at the 5% level with correlation value of 99.96% was also obtained.

The Effect of Variable Incubation Temperatures on Hatchability and Survival of Goldlined Seabream, Rhabdosargus sarba (Forsskål,1775) Larvae

The effect of varying holding temperature on hatching success, occurrence of deformities and mortality rates were investigated for goldlined seabream eggs. Wild broodstock (600 g) were stocked at a 2:1 male-female ratio in a 2 m3 fiberglass tank supplied with filtered seawater (37 g L-1 salinity, temp. range 24±0.5 oC [day] and 22±1 oC [night], DO2 in excess of 5.0mg L-1). Females were injected with 200 IU kg-1 HCG between 08.00 and 10.00 h and returned to tanks to spawn following which eggs were collected by hand using a 100μm net. Fertilized eggs at the gastrulation stage (120 L-1) were randomly placed into one of 12 experimental 6 L aerated (DO2 5 mg L-1) plastic containers with water temperatures maintained at 24±0.5 oC (ambient), 26±0.5 oC, 28± 0.5 oC and 30±0.5 oC using thermostats. Each treatment was undertaken in triplicate using a 12:12 photophase:scotophase photoperiod. No differences were recorded between eggs reared at 24 and 26 oC with respect to viability, deformity, mortality or unhatched egg rates. Increasing temperature reduced the number of viable eggs with those at 30 oC returning poorest performance (P < 0.05). Mortality levels were lowest for eggs incubated at 24 and 26 oC. The greatest level of deformities recorded was that for eggs reared at 28 oC.

Effect of Fermentation Time on Xanthan Gum Production from Sugar Beet Molasses

Xanthan gum is a microbial polysaccharide of great commercial significance. The purpose of this study was to select the optimum fermentation time for xanthan gum production by Xanthomonas campestris (NRRL-B-1459) using 10% sugar beet molasses as a carbon source. The pre-heating of sugar beet molasses and the supplementation of the medium were investigated in order to improve xanthan gum production. Maximum xanthan gum production in fermentation media (9.02 g/l) was observed after 4 days shaking incubation at 25°C and 240 rpm agitation speed. A solution of 10% sucrose was used as a control medium. Results indicated that the optimum period for xanthan gum production in this condition was 4 days.

New Device for Enhancement of Liposomal Magnetofection Efficiency of Cancer Cells

Liposomal magnetofection is the most powerful nonviral method for the nucleic acid delivery into the cultured cancer cells and widely used for in vitro applications. Use of the static magnetic field condition may result in non-uniform distribution of aggregate complexes on the surface of cultured cells. To prevent this, we developed the new device which allows to concentrate aggregate complexes under dynamic magnetic field, assisting more contact of these complexes with cellular membrane and, possibly, stimulating endocytosis. Newly developed device for magnetofection under dynamic gradient magnetic field, “DynaFECTOR", was used to compare transfection efficiency of human liver hepatocellular carcinoma cell line HepG2 with that obtained by lipofection and magnetofection. The effect of two parameters on transfection efficiency, incubation time under dynamic magnetic field and rotation frequency of magnet, was estimated. Liposomal magnetofection under dynamic gradient magnetic field showed the highest transfection efficiency for HepG2 cells.