Characterization and expression of the pseudorabies virus (NYJ strain) glycoproteins in Bombyx mori cells and larvae

To characterize the NYJ strain of pseudorabies virus (PRV; Alphaherpesvirus of swine) isolated from the serum of an infected swine in Korea, the nucleotide sequence of three major glycoproteins (gB, gC, and gD) was analyzed. The expression of most potent immunogenic glycoprotein (gD) was also investigated using a Bombyx mori nucleopolyhedrovirus (BmNPV) expression system. The length of the glycoprotein genes corresponding to gB, gC, and gD of the NYJ strain were 2751 bp, 1443 bp, and 1203, respectively, and their identity ranged from 94.2% to 99.8% when compared with other strains. Phylogenetic analyses using these sequences showed that the NYJ strain forms a distinct branch with high bootstrap support. A novel transfer vector (pBmKSK4) was engineered with the polyhedrin promoter of BmNPV and a 6xHis tag to express glycoprotein gD in Bm5 cells and silkworm, B. mori, larvae. The immunogenicity of recombinant gD was demonstrated by its specific detection in both Bm5 cells and silkworm larvae by porcine anti-PRV antibody. The results of this study have implications both for the taxonomy of Korean PRV strains and vaccine development.     

Oil production from five marine microalgae for the production of biodiesel

Marine microalgae were studied as potential resources for the production of biodiesel. Five marine microalgae, Tetraselmis suecica, Phaeodactylum tricornutum, Chaetoceros calcitrans, Isochrysis galbana, and Nannochloropsis oculata were cultured in f/2 media, 12:12 L:D cycle at 20 ± 1°C with a light intensity of 36.3 μmol/m²/sec using a 15-L circular cylindrical photobioreactor. The dry cell weight, specific growth rate, biomass productivity, oil content and fatty acid composition of palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid of microalgae were determined. T. suecica, I. galbana, and N. oculata showed high dry cell weights of 0.58, 0.57, and 0.57 g/L, respectively. The culture period of T. suecica to reach the stationary phase was 9 days. On the other hand, N. oculata showed the longest culture period of 28 days to reach the stationary phase. T. suecica absorbed nitrate at the initial stages of cell growth, decreasing the nitrate concentration to 0.5 mg/L on day-7 of the culture. The highest oil contents were observed in P. tricornutum with a 25.31% dry cell weight and I. galbana with a 23.15% dry cell weight on day-9 after the stationary phase. I. galbana showed 417.33 mg of palmitic acid per g oil and T. suecica showed 235.61 mg of oleic acid per g oil. Stearic acid, linoleic acid, and linolenic acid did not exceed 30.02 mg/g oil in any of the microalgae. T. suecica showed the shortest culture period of 9 days to reach the stationary phase. Therefore, the highest biomass production of 0.58 g/L was obtained and I. galbana showed high biomass production of 0.57 g/ L and oil content of 23.15% of dry cell weight. Therefore, T. suecica and I. galbana can be selected as a potential candidate for the production of biodiesel.     

Distribution of Sunn Pest,Eurygaster integriceps Puton (Hemiptera: Scutelleridae), in overwintering sites

Sunn Pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), is the most serious pest of wheat in West and Central Asia. Following harvest, the pest migrates from wheat fields to high elevations to escape high summer temperatures and, the following spring, moves back to the fields. In this work, Sunn Pest populations were monitored in 21 overwintering sites located in seven provinces (with 800–2500 m elevation) in Iran. Litter samples were taken twice in each site, once in July–November 2002 and again in March 2003. At each site 60 Artemesia sp. bushes were selected at random and litter from each bush was collected and the number of live male and female adult Sunn Pest counted. More Sunn Pests were collected in higher elevations in the first sampling. Golestan (893 m), Kermanshah (1583 m), and Qazvin (1675 m), located in valleys and surrounded by mountainous areas, had < 5 adults/bush. However Azarbayejan (1750 m), Markazi (1883 m), Kordestan (2067 m), and Esfahan (2141 m), located in mountainous areas and/or close to mountains, had 10–52 adults/bush. No noticeable changes in Sunn Pest populations were observed in 17 overwintering sites among 21 between the two sampling times. These results suggest that, after Sunn Pest moves to more highly elevated sites at aestivation time, most of them remain in these sites during overwintering before migration to wheat fields in the spring. Sunn Pest can be targeted for management in these areas during the fall and/or early spring as their movement is clearly limited.     

Highly expressed cholera toxin B subunit in the fruit of a transgenic tomato (<Emphasis Type="Italic">Lycopersicon esculentum</Emphasis> L.)

The cholera toxin B subunit (CTB), a nontoxic molecule with potent biological properties, is a powerful mucosal and parenteral adjuvant that induces a strong immune response against co-administered or coupled antigens. A gene encoding CTB, which was modified based on the optimized codon usage in the plant, was synthesized and fused to the endoplasmic reticulum retention signal KDEL to enhance its expression level in plants. The synthetic CTB (sCTB) gene was introduced into a plant expression vector adjacent to the CaMV 35S promoter, and was transformed into tomato using an Agrobacterium-mediated transformation method. The integration of the sCTB gene into the genomic DNA of transgenic plants was confirmed by genomic DNA PCR amplification. The synthesis and assembly of CTB protein in transgenic plants was demonstrated through immunoblot analysis and G_M1-ELISA. The highest amount of CTB protein produced in transgenic tomatoes was approximately 0.9% of total soluble fruit protein which was 10-fold greater than the previously 0.081%. G_M1-ELISA indicated that plant-synthesized CTB protein bound specifically to G_M1-gangliosides, suggesting that the CTB subunits formed active pentamers.     

Statistical optimization of fermentation conditions and comparison of their influences on production of cellulases by a psychrophilic marine bacterium, <Emphasis Type="Italic">Psychrobacter aquimaris</Emphasis> LBH-10 using orthogonal array method

The optimal conditions for the production of cellulases by a marine bacterium, Psychrobacter aquimaris LBH-10, were established and their effects were compared using orthogonal array experiments based on the Taguchi method. The optimal conditions of rice bran, peptone and initial pH for the production of avicelase and CMCase by P. aquimaris LBH-10 were 50.0, 3.0, and 8.0 g/L, respectively, whereas those for filter paperase (FPase) were 100.0, 3.0, and 8.0 g/L, respectively. Rice bran was found to be the most important factor for the production of cellulases based on the calculated percentage of participation P (%) from an analysis of the variance (ANOVA). The optimal temperature for the cell growth of P. aquimaris LBH-10 was 25°C, whereas that for the production of avicelase, CMCase and FPase was 30°C. The optimal agitation speed and aeration rate for cell growth was 400 rpm and 1.5 vvm, respectively, whereas those for the production of CMCase were 300 rpm and 1.0 vvm, respectively. Aeration was found to be more important for cell growth and CMCase production than agitation. The maximum production of avicelase, CMCase and FPase in a 100 L bioreactor for 72 h under optimized conditions was 83.2, 388.7, and 75.4 U/mL, respectively.     

Real-time monitoring of oriental fruit moth,Grapholita molesta,populations using a remote sensing pheromone trap in apple orchards

A fusion of information technology (IT) and sex pheromone monitoring provides a remote sensing IT-pheromone trap to monitor Oriental fruit moth, Grapholita molesta, populations in apple orchards. Once a male of G. molesta is attracted to its sex pheromone lure in the trap, an infrared sensor installed at the funnel-shaped orifice generates an electric signal. The signal is processed in a central processor and then transferred to an internet site via a code division multiple access protocol. The signal also contains information about when each male is caught. Daily trapping information from different localities is archived in a website. The accuracy of IT-pheromone traps in detecting male catches was shown by a high correlation (r = 0.956) between the generated IT signals and actual numbers of males caught in the trap in apple orchards. Using this IT-pheromone trap, G. molesta in apple orchards was monitored for one year. These data were compared with monitoring data obtained from a conventional wing type-based sticky trap containing the identical sex pheromone lure. Both showed four characteristic adult peaks from April to September and were significantly correlated (r = 0.695). IT-pheromone traps also gave real-time signals of male catches in the field. These real-time signals of male catches showed a characteristic diel attraction rhythm from 4 pm to midnight. The diel rhythm of the male response to the sex pheromone started earlier in the evening in the spring season compared to mid and late seasons. This study provides a novel sex pheromone trap for G. molesta to monitor its population in field conditions in real-time without visiting or counting. The field monitoring data can be accessed any time through a designated internet website.     

Immunostimulatory effect by aqueous extract of <Emphasis Type="Italic">Hizikia fusiforme</Emphasis> in RAW 264.7 macrophage and whole spleen cells

Hizikia fusiforme is a commonly used food that possesses potent anti-bacterial, anti-fungal, and anti-inflammatory activities. The immunostimulatory activities of aqueous extract of Hizikia fusiforme (HFAE) in RAW 264.7 macrophages and whole spleen cells were investigated. HFAE activated RAW 264.7 macrophages to produce cytokines such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in a dose-dependent manner. In addition, HFAE induced the mRNA expression of TNF-α, IL-1β, and IL-6 in RAW 264.7 macrophages. Moreover, HFAE stimulated proliferation of whole spleen cells and reference mitogen. Taken together, the results demonstrate that HFAE potently activates the immune function by regulating NO, TNF-α, IL-1β, and IL-6 in RAW 264.7 macrophage and promoting spleen cell proliferation.     

Potential agents for cancer and obesity treatment with herbal medicines from the green garden

Many classes of bioactive drug-like molecules derived from traditional herbal plants are becoming attractive as alternative medicines for the treatment of severe chronic diseases such as cancer and obesity. A set of chemically synthesized drugs that is capable of both inhibiting cancer growth and reducing body weight for treatment of obesity have severe side effects including nausea, vomiting, diarrhea as well as producing increased blood pressure and headache, respectively. For decades, drug candidates from herbal plants have been considered as potential therapeutic agents because they are generally safer, less toxic, and have fewer lethal side effects than chemically synthesized or semi-synthetic drugs. Understanding the key factors affecting pharmacological effects and clinical outcomes has been a critical theme of natural product research. However, standardized sample preparation methods, well-controlled scientific studies, and validation studies are needed before herbal therapeutics can be introduced into the global market. This review will address the current advances in using traditional herbal plants, including the pharmacological effects and the challenges faced during the development of new drugs. The safety issues associated with toxicity and the effectiveness of the herbs in specific diseases such as cancer and obesity are also discussed.     

Insecticide susceptibility of Ephemera orientalis (Ephemeroptera: Ephemeridae) and two mosquito species,Anopheles sinensis and Culex pipiens in the Republic of Korea

Field-collected populations of mayflies, Ephemera orientalis were tested for susceptibility to 10 different insecticides using a direct-contact mortality bioassay. Ephemera orientalis subimagoes were susceptible to the insecticides chlorpyrifos, fenitrothion and chlorfenapyr with LD₅₀ values of 69.7, 78.8 and 81.9 μg/♀, and adults had LD₅₀ values of 71.9, 78.8 and 85.4 μg/♀, respectively. Susceptibility ratios (SRs) of subimagoes and adults of E. orientalis to the 10 insecticides were 1.0 to1.2 folds. The mayflies showed higher susceptibility to organophosphates than to pyrethroids. The SRs of Anopheles sinensis to E. orientalis were 514 to 1438 folds higher for organophosphates (LD₅₀ values of 0.05 to 0.23 μg/♀) and 62 to 1155 folds higher for pyrethroids (LD₅₀ values of 0.13 to 2.41 μg/♀). The SRs of Culex pipiens to E. orientalis were 606 to 3595 folds higher for organophosphates with LD₅₀ values of 0.02–0.17 μg/♀ and 81 to 1365 folds higher for pyrethroids with LD₅₀ values of 0.11–1.83 μg/♀. These results indicate that the use of ineffective insecticides will result in unsatisfactory control against field populations of the subimagoes and adults of E. orientalis.