The objectives of this study were to evaluate the antimicrobial effects of ε-poly-L-lysine mixture (EPM) on cheon-ggukjang and to determine the optimal mixing ratio in cheonggukjang. Increasing the EPM up to 2% decreased via-ble cell counts at both 15oC and 30oC storage. The pH of cheonggukjang without EPM increased slowly over thestorage periods, but cheonggukjang with EPM showed a slight increasing tendency, with the highest pH at 2% EPM.The sensory scores, such as texture and flavor, were highest in samples containing 2% EPM; however, overall pref-erence was not significantly different when compared to the control. For the optimum cheonggukjang processingconditions, 13 experimental points were selected. Soybean and EPM were chosen as independent variables. Viablecell count, pH, texture, and overall preference were measured. The optimum formulation of cheonggukjang usingthe numerical analysis was found to be 98.52% soybean and 1.48% EPM, resulting in a 0.722 desirability value.

The cerebellum is known to control balance, equilibrium, and muscle tone. If the cerebellum becomes damaged, the body is unable to retain its balancing functions or involuntary muscle movement. This is why, in stroke patients, there is a high risk of functional disability, as well as a myriad of other disabilities secondary to stroke. Ischemia was induced in SD mice by occluding the common carotid artery for 5 minutes, after which blood was reperfused. Needle electrode electrical stimulation(NEES) was applied to acupuncture points, at 12, 24, and 48 hours post-ischemia on the joksamri. Protein expression was investigated through caspase-3 antibody immuno-reactive cells in the cerebral nerve cells and Western blotting. The results were as follows: The number of caspase-3 reactive cells in the corpus cerebellum 12 and 24 hours post-ischemia was significantly (p<.05) smaller in the NEES group compared to the GI group. caspase-3 expression 12 and 24 hours post-ischemia was significantly(p<.05) smaller in the NEES group compared to the GI group. Based on these results, NEES seems to have a significant effect on Caspase-3 in the cerebellum in an ischemic state at 12 and 24 hours post ischemia, NEES delays the occurrence of early stage apoptosis-inducing Caspase-3, delaying and inhibiting apoptosis. Further systematic studies will have to be conducted in relation to the application of this study’s results on stroke patients.

Insect pollinators of the endanger orchid Cypripedium japonicum were surveyed and identified during two years, as a part of a conservation project of the orchid at Jukyeup-san and Hwaak-san (Mt.), South Korea. In total 40 individuals of 16 species in 4 families were identified. The dominant family was Halictidae, and Lasioglossum exiliceps Vachal visited the most frequently C. japonicum during the surveys. The average visiting frequency was 2.5 individuals per hour and the highest 4.3, from 12:00 – 13:00 in a day. After 15:00 insects did not visit the flowers at all. However, all of the visiting insects were found to not carry a pollinium or pollens of the orchard on their bodies; pollen carryover by any of the visiting insects did not occur at all. The orchid seems to require certain pollinators in particular body thickness due to its unique pollination mechanism. The orchid has two exit route openings, around 1 cm in diametre, where the entrapped insects can exit and an anther is situated just in front of each opening. It was inferred that a pollen carrier should be around 1 cm in body thickness. Therefore, the candidate species as the proper pollen carriers can be Tetralonia nipponensis Perez, Xylocopa appendiculata circumvolans Smith and Bombus consobrinus Dahlbom among the surveyed visitors.

Mushrooms are considered not only as food but also for source of physiologically beneficial medicines. The culinarymedicinal mushrooms may important role in the prevention of age-associated neurological dysfunctions, including Alzheimer’s and Parkinson’s diseases. Hericium erinaceus (H. erinaceus), is edible mushrooms, is a parasitic fungus that grows hanging off of logs and trees and well established candidate for brain and nerve health. H. erinaceus contains high amounts of antioxidants, beta-glucan, polysaccharides and a potent catalyst for brain tissue regeneration and helps to improve memory and cognitive functions. Its fruiting bodies and the fungal mycelia exhibit various pharmacological activities, including the enhancement of the immune system, antitumor, hypoglycemic and anti-aging properties. H. erinaceus stimulates the synthesis of Nerve Growth Factor (NGF) which is the primary protein nutrient responsible for enhancing and repairing neurological disorders. Especially hericenones and erinacines isolated from its fruitin body stimulate NGF, synthesis. This fungus is also utilized to regulate blood levels of glucose, triglycerides and cholesterol. H. erinaceus can be considered as useful therapeutic agents in the management and/or treatment of neurodegeneration diseases. However, this review focuses on in vitro, in vivo and clinical trials for neurodegerative disease.

The antibiotic peptide PAJE (RWKIFKKPFKISIHL-NH2), designed incorporating the N-terminal α-helical segments of papiliocin and jelleine, is a 15-residue hybrid peptide that has a broad spectrum of activity against Gram-negative, positive bacteria and fungi. In this study, we successfully expressed bioactive PAJE in Escherichia coli cells that are highly sensitive to this peptide. For the efficient production of peptide, we synthesized gene encoding PAJE, and fused the sequence in-frame to ketosteroid isomerase (KSI) gene to construct an expression vector pET29b-PAJE-KSI, which was then used to transform E. coli BL21 (DE3). The fusion protein PAJE-KSI was expressed as inclusion body at high level (more than 30% of the total proteins). Recombinant PAJE was easily released by cleavage of the fusion protein with cyanogen bromide (CNBr). Subsequently, we purified the recombinant PAJE by FPLC chromatography. The purified PAJE displayed considerably antibacterial activity identical to that previously reported for chemically synthesized PAJE. The results indicated that successful expression of PAJE in E. coli cells and efficient procedure for purification may lead to a cost-effective platform for the mass production of PAJE.

Antimicrobial peptides (AMPs) can be produced in mealworms, currently being used as animal feeds, by the infection of genetically engineered-entomopathogenic fungi. In this work, we integrated Bombyx mori (Bm) AMP, cecropin A to Beauveria bassiana ERL1170 by restriction enzyme-mediated integration method, which was confirmed by RT-PCR and an antibacterial activity assay. For the extracellular secretion of Bm cecropin A protein, the active domain of the cecropin A gene was tailed to the signal sequence of B. bassiana chitinase (Bbs). To exchange Bbs-cecropin A gene with egfp gene in pBARKS1-egfp, Bbs-cecropin A fragment was cut from pGEM-Bbs-cecropin A using XbaI/blunted and BamHI and ligated with cut pBARKS1-egfp using NcoI/blunted and BamHI, designated as pBARKS1-Bbscecropin A. After the transformation, transformants were grown on Czapek’s solution agar containing 600 μg ml-1PPT. Expression of Bm cecropin A was confirmed by RT-PCR. Strong clear zone was observed in the co-culture of the transformant D-6 and Bacillus subtilis on fourth strength Sabouraud dextrose agar 1 day after the culture at 25°C, whereas the wild type had no clear zone. This work suggests that Bm cecropin A can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

Brown planthopper, Nilaparvata lugens is giving enormous damage to rice production. In this work, virulence of four Beauveria bassiana isolates against brown planthoppers was investigated by applying fungal granules on the water of pots, and we further examined the growth of hypha on the rice plants from the water surcae to explain the insecticidal mode of action. We used Beauveria bassiana (Bb) ERL 836, 1170, 1575 and 1578 isolates, which produced ~2×108 conidia/g of millet grain in a solid culture. Rice seeding was grown in breeding boxes at 28±2℃ for 5 days. Mycotized millet grains were treated on the water in a box at 1 g/box and the rice seeding was infested with 18~25 brown planthopper adults per box. A chemical pesticide and water treatment served as controls. Among the treatments, Bb ERL 836-treated plants had the lowest damage, rather than the other fungal treatments in laboratory assays. Hyphal growth on the stem of rice plants was observed in Petridish conditions under a fluorescent microscopy. This work suggests a possible control of brown planthoppers using entomopathogenic fungi.

It is true that the proper environmental risk assessments for many GM insects almost have not been executed in Korean research situation. Therefore, we tested the environmental risk assessment of GM silkworms if there is any difference between GM and non-GM silkworms by three. First, we measured their mobility in the breeding environment conditions with food and without food. Secondly, we measured their viability at the Korean artificial extreme environmental conditions (temperature, humidity, food) after escaping from their breeding environments. We observed the egg productivity and the hatchability between non-GM silkworm and transgenic silkworms with four different pair experiments. The mobility of non-GM silkworms and GM silkworms statistically did not differ and the egg production and hatchability was not also different. The hatchability by couple of GM female silkworm and non-GM male silkworm was lower than by non-GM male and female couple. We observed their viability (High Temp., wet and with food: p=0.0434; High Temp., wet and without food p=0.0430; High Temp., dry and with food: p=0.0005; High Temp., dry, without food: p=0.0479) between the GM silkworm and non-GM silkworm, and there was statistically different. Relatively, the viability of GM silkworm was lower than non-GM silkworms. We could not exactly test for viability of silkworm in low temperature conditions because of their hibernating. Although there was any difference in viability and hatchability between GM silkworm and non-GM silkworm, the all ability of GM silkworm was lower than non-GM silkworm. Conclusively, risk of GM silkworm was lower than non-GM silkworm.

Post-maternal social organisation of Amaurobius ferox (Araneae, Amaurobiidae) provides an opportunity to investigate the mechanisms of social behaviour in spiderlings while controlling for potential effects of maternal presence. Development and characteristics of locomotory behaviours in young individuals of the subsocial Amaurobius ferox were examined. Young collectively show a very rapid and linear locomotion in a short distance in response to an intruder in the natal nest. Such a locomotory behaviour of the spiderlings resembles the successive synchronised stepping towards prey of the permanently social spider, Anelosimus eximius. We observed the development of different types of locomotion relative to the age of young, collective tendency (i.e., proportion of individuals within the clutch engaging in the behaviour), and directional properties of the locomotion. Locomotory behaviours of spiderlings fell into two categories: ‘abrupt locomotion’ (AL), involving fast and linear movement over a short distance; and ‘ordinary locomotion’ (OL), involving slower movement over longer distances and times. The frequency of AL (abrupt locomotion) did not vary much throughout the days post-matriphagy, occurring for a limited period of time. On the other hand, the frequency of OL (ordinary locomotion) varied over the days post-matriphagy and increased during the maternal social period.

Beauveria bassiana isolates have been used in integrated pest management, but little consideration has been given to the studies on fungal gene expressions and their functions. In this work, to determine the functions of genes, B. bassiana ERL1170 was transformed by restriction enzyme-mediated integration method, where pABeG with bar gene was used as a transformation vector. Among seven hundred of transformants, morphologically different ERL1170-pABeG-#160 transformant, particularly dysfunctional in conidiogenesis. The transformant had yellow hyphal growth on fourth strength Sabouraud dextrose agar (SDA/4) and produced very small amount of conidia (<1.0×105 conidia/cm2 agar) in 7 days, whereas wild type had white mycelial growth and significantly greater conidia (3.6×106 conidia/cm2 agar). Additionally under microscopic observation, hyphae of #160 seemed like indian club, compared to the straight forms of wild type hyphae. The next work is figure out possible genes contributing the conidiogenesis of B. bassiana.

Entomopathogenic fungus, Beauveria bassiana can be used in integrated crop management and pharmaceutical applications. Recently some efforts have been given to the fungus to improve its biological performances, but low fungal transformation efficiency is one of the limitations. In this work, B. bassiana ERL1170 isolate was used for fungal transformation by restriction enzyme-mediated integration, where pBARKS1-Bbs-cecropinA was linearied using HindIII. The fungal transformation comprised two steps, preparation of competent blastospores and integration of the plasmid into the cells. To prepare competent blastospores, 2-d cultured blastospores were individually treated with 0.1, 0.2, 0.4 and 0.8 M lithium acetate (LiAc). Secondly in the integration step, concentration of LiAc and calcium chloride (CaCl2), and time period of heat shock were investigated as follows: LiAc, 1, 2 and 4 M; CaCl2, 50, 100 and 200 mM; and heat shock at 42℃, 20, 40 and 60 min. Consequently, combination of 0.2 M LiAc in preparing competent blastospores and 2 M LiAc and 200 mM CaCl2 in the second step showed the highest transformation efficiency. This work would be helpful in the fungal transformation of B. bassiana.

Ischemia, the leading cause of strokes, is known to be deeply related to synaptic plasticity and apoptosis in tissue damage due to ischemic conditions or trauma. The purpose of this study was to research the effects of NEES(needle electrode electrical stimulation) in brain cells of ischemiainduced rat, more specifically the effects of Poly[ADP-ribose] polymerase(PARP) on the corpus striatum. Ischemia was induced in SD mice by occluding the common carotid artery for 5 minutes, after which blood was re-perfused. NEES was applied to acupuncture points, at 12, 24, and 48 hours post-ischemia on the joksamri, and at 24 hours postischemia on the hapgok. Protein expression was investigated through PARP antibody immuno-reactive cells in the cerebral nerve cells and western blotting. The number of PARP reactive cells in the corpus striatum 24 hours post-ischemia was significantly(p<.05) smaller in the NEES group compared to the global ischemia(GI) group. PARP expression 24 hours post-ischemia was very significantly smaller in the NEES group compared to the GI group. Results show that ischemia increases PARP expression and stimulates necrosis, making it a leading cause of death of nerve cells. NEES can decrease protein expression related to cell death, protecting neurons and preventing neuronal apoptosis.

The nasal cavity encounters various irritants during inha¬lation such as dust and pathogens. To detect and remove these irritants, it has been postulated that the nasal mucosa epithelium has a specialized sensing system. The oral cavity, on the other hand, is known to have bitter taste receptors (T2Rs) that can detect harmful substances to prevent ingestion. Recently, solitary chemosensory cells expressing T2R subtypes have been found in the respiratory epithelium of rodents. In addition, T2Rs have been identified in the human airway epithelia. However, it is not clear which T2Rs are expressed in the human nasal mucosa epithelium and whether they mediate the removal of foreign materials through increased cilia movement. In our current study, we show that human T2R receptors indeed function also in the nasal mucosa epithelium. Our RT-PCR data indicate that the T2R subtypes (T2R3, T2R4, T2R5, T2R10, T2R13, T2R14, T2R39, T2R43, T2R44, T2R 45, T2R46, T2R47, T2R48, T2R49, and T2R50) are expressed in human nasal mucosa. Furthermore, we have found that T2R receptor activators such as bitter chemicals augments the ciliary beating frequency. Our results thus demonstrate that T2Rs are likely to function in the cleanup of inhaled dust and pathogens by increasing ciliary movement. This would suggest that T2Rs are feasible molecular targets for the development of novel treatment strategies for nasal infection and inflammation.

Opioid receptors have been pharmacologically classified as µ, δ, κ and ε. We have recently reported that the antinociceptive effect of morphine (a µ-opioid receptor agonist), but not that of β-endorphin (a novel µ/ε-opioid receptor agonist), is attenuated by whole body irradiation (WBI). It is unclear at present whether WBI has differential effects on the antinociceptive effects of µ-, δ-, κ- and ε-opioid receptor agonists. In our current experiments, male ICR mice were exposed to WBI (5Gy) from a 60 Co gamma-source and the antinociceptive effects of opioid receptor agonists were assessed two hours later using the hot water (52℃) tail-immersion test. Morphine and D-Ala2,N-Me-Phe4,Gly-ol-enkephalin (DAMGO), [D-Pen2-D-Pen5]enkephalin (DPDPE), trans-3,4-Dichloro-N-methyl-N-[2-(1-pyrrolidinyl)- cyclohexyl]¬benzeneacetamide (U50,488H), and β-endorphin were tested as agonists for µ, δ, κ, and ε-opioid receptors, respectively. WBI significantly attenuated the antinociceptive effects of morphine and DAMGO, but increased those of β-endorphin. The antinociceptive effects of DPDPE and U50,488H were not affected by WBI. In addition, to more preciously understand the differential effects of WBI on µ- and ε¬opioid receptor agonists, we assessed pretreatment effects of β-funaltrexamine (β-FNA, a µ-opioid receptor antagonist) or β-endorphin1-27 (β-EP1-27, an ε-opioid receptor antagonist), and found that pretreatment with β-FNA significantly attenuated the antinociceptive effects of morphine and β endorphin by WBI. significantly reversed the β-EP1-27 attenuation of morphine by WBI and significantly attenuated the increased effects of β-endorphin by WBI. The results demonstrate differential sensitivities of opioid receptors to WBI, especially for µ- and ε-opioid receptors.

The purpose of this study was to investigate the influence of handicraft activities on hand promptness and grasp in the elderly. Subjects were comprised of 14 senior citizens between the ages of 70-85, with 7 subjects in the experiment group and 7 in the control group. Subjects in the experiment group practiced various handicrafts twice a day, while those in the control group did not participate in any special activity. The Jebsen Taylor Hand Function Test was used to evaluate the results, while a dynamometer and pinch gauge were used to measure hand promptness and grasp. The 7 senior citizens in the experiment group were able to increase their hand promptness and grasping skills. Conclusively, handicrafts can help improve hand promptness and grasp in the elderly. Furthermore, the development and improvement of such skills can have a positive influence on the daily lives of senior citizens. Such skills are expected to improve the overall neuro-function in the elderly population.

The leaf beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) of the species collected from seven Korean localities. A total of 17 haplotypes (CACOI01 ~ CACOI17), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 16 sequence types (ITS2CA01 ~ ITS2CA16), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in COI gene sequence. Phylogenetically, the COI gene provided two haplotype groups with a high nodal support (≥ 87%), whereas ITS2 provided one sequence type group with a high nodal support (≥ 92%). The result of COI gene may suggest the presence of historical biogeographic barriers that bolster genetic subdivision in the species. Different grouping pattern between COI gene and ITS2 sequences were interpreted in terms of recent dispersal, reflected in the ITS2 sequence. Finally, finding of unique haplotypes and sequence types only from Beakryeng-Islet population was interpreted as an intact remnant of ancient polymorphism. As more samples are analyzed using further hyper-variable marker, further fruitful inference on the geographic contour of the species might be available.