• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.17-25
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• 2013

The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.

• Journal of fish pathology
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• v.2 no.1
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• pp.1-18
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• 1989

In Korea, studies on a Nematode, Anguillicola crassa parasitic in the air bladder of eel are not yet reported. This reason led the author to study the parasitic species, state and life history of the A. crassa parasitized in the air bladder of eel in order to take effective control measures against its damage. The size of fully developed eggs was 80 to $92(86.7){\times}62$ to $71(67.4)\;{\mu}m$, larva was 210 to $240(225){\times}18$ to $23(20.6)\;{\mu}m$. The intermediate host of A. crassa was Thermocyclops hyalinus, it was capable for parasitizing the eel after 4 days of invasion and then the size of larva was 360 to $420(390){\times}28$ to $35(31)\;{\mu}m$. Fifty days after eel had ingested the Thermocyclops hyalinus infected with larva of A. crassa, the larvae matured into adult worms in the air bladder of eel. The size of detected adult worms was 7.3 to $31.0(16.5){\times}0.5$ to 2.2(1.2) mm, 4.9 to $13.3(8.3){\times}0.3$ to 0.9(0.4) mm. Investigating the morphology of the worms, they were identified as A. crassa. Monthly the parasitic rate of the worms in the eel was high in June, September and December, but low in January to March. After the investigation on the significance between non-parasitic fish and parasitic fish, it was not significant, therefore it can be considered that there is no effect of infection in the growth of eel. Any abnormality of eels air bladder tissue was not seen by the infection of A. crassa. At 25.0 to $26.7^{\circ}C$ of water temperature the death time of Thermocyclops hyalinus by masoten treatment was 14 hours in 0.5 ppm, 20 hours in 0.4 ppm, 22 hours in 0.3 ppm, 30 hours in 0.2 ppm and 42 hours in 0.1 ppm.

• Toxicological Research
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• v.8 no.2
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

• Journal of Life Science
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• v.24 no.1
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• pp.86-91
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• 2014

Trichoplusia ni cells are used as a host permissive cell line in the baculovirus expression system, which is useful for large-scale production of human sugar transport proteins. However, the activity of endogenous sugar transport systems in insect cells is extremely high. Therefore, the transport activity resulting from the expression of exogenous transporters is difficult to detect. Furthermore, very little is known about the nature of endogenous insect transporters. To exploit the expression system further, the effect of D-fructose on 2-deoxy-D-glucose (2dGlc) transport by T. ni cells was investigated, and T. ni cell-expressed human transporters were photolabeled with [$^3H$] cytochalasin B to develop a convenient method for measuring the biological activity of insect cell-expressed transporters. The uptake of 1 mM 2dGlc by uninfected- and recombinant AcMPV-GTL infected cells was examined in the presence and absence of 300 mM of D-fructose, with and without $20{\mu}M$ of cytochalasin B. The sugar uptake in the uninfected cells was strongly inhibited by fructose but only poorly inhibited by cytochalasin B. Interestingly, the AcMPV-GTL-infected cells showed an essentially identical pattern of transport inhibition, and the rate of 2dGlc uptake was somewhat less than that seen in the non-infected cells. In addition, a sharply labeled peak was produced only in the AcMPV-GTL-infected membranes labeled with [$^3H$] cytochalasin B in the presence of L-glucose. No peak of labeling was seen in the membranes prepared from the uninfected cells. Furthermore, photolabeling of the expressed protein was completely inhibited by the presence of D-glucose, demonstrating the stereoselectivity of labeling.

• Korean Journal of Food Science and Technology
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• v.35 no.1
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• pp.15-22
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• 2003

Nutritional compositions of three Tochukaso species (Paecilomyces tenuipes hosted by Larva and pupa, Cordyceps militaris, C. sinensis) were compared. Fruiting body and host fractions were separately analyzed. Fruiting body fraction of P. tenuipes (36.6%) hosted by larva was higher than that hosted by pupa (10.2%), an indication that the quality of the former is superior to the latter. Carbohydrate content of C. sinensis (39.6%) was $2.5{\sim}7$ times higher than those of others, probably due to the presence of polysaccharides. Protein and crude lipid contents of C. sinensis and C. militaris were 25.8 and 10.3%, and 75.1 and 3.9%, respectively. C. sinensis showed the lowest Ca content and $30{\sim}75$ times higher Fe content among the samples tested. Vitamin A content of C. militaris was 308.9 IU/100g, two fold higher than those of the other species. Saturated fatty acid content was the highest in P. tenuipes (pupa, 27.7%), whereas unsaturated fatty acid was the highest in P. tenuipes (larva, 83.3%). Aspartic acid, glutamic acid, and glycine were abundant in all species. Cordycepin content of C. militaris was $20{\sim}50$ times higher than those of the other species.

• Journal of the Speleological Society of Korea
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• no.95
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• pp.35-49
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• 2009

Ever since the beginning of time, caves not only have offered a place to live for humans but they have also been used as cultural spaces. That is, in the event of making some sounds in a location within the cave, the sound that is created is greatly magnified and sounds out as if it is being amplified from a giant megaphone. This, as we well know it, is known as the resonance effect. Here, the cave itself appears to function as a massive wind instrument. Especially in cases like the Altamira Cave (Spain) where cave paintings were found, the point where the cave drawings were found has commonalities in that it is a wide space and that it is usually discovered together with flutes and drums that are made with mammoth bones. We need to focus on this point. We can infer from these facts that the prehistoric people have carried out cultural activities along with their incantation rituals within those caves. In the meantime, amongst the Korean traditional arts, in the case of pansori which is a representative vocal genre, there have been examples where caves were used as practicing locations for those people who are training to perfect their singing. This is known as toguldoggong(土窟獨功) which literally means 'obtaining one's own art by oneself in the earth cave by practicing incessantly'. This process along with pokpodoggong (瀑布獨功) (same as above except that the location is by the waterfall) is the final training stage in order to become a recognized virtuoso on the part of the apprentice. This could be compared to the final annealing and finishing process of producing a metalwork. This has been a long tradition followed by most Korean traditional artists in order to perfect their sound which is harmonious with nature within natural surroundings. By honing in on this point, I have come to think about this matter repeatedly while coaching the university students in vocal singing. In short, I came to the conclusion that "the making of natural sounds will be obtained naturally within natural surroundings like caves!" Consequently, The Society for Studying Cave Sounds was inaugurated on January 1992 along with some of my students. We made use of times like vacations to go around exploring caves all over Jeju and carried out investigations of sounds along with cave exploration on an experimental basis. After 5 years, in September of 1997, we were able to host the first ever cave concert domestically at the Whale Nostril Cave(東岸鯨窟) on Wu-do. After that, we have been hosting the cave concert once every year. We have achieved a record of a total of 14 cave concerts until 2009 of this year. Out of these, 2 were held in Seokhwaeam Cave in Kangwon Province, another two were held in Manjang Cave which is a lava cave, and the remaining 10 were held in the Whale Nostril Cave of Wu-do. Along with that, I have carried out a special recording for the production of a cave music CD in May of 1999. This paper was written and organized by using the main materials that were derived from the experiences of using caves as concert halls in the past. It is hoped that this cave concert will offer a very unique experience to tourists who come to Jeju every year and give them the best possible superior natural sound effect that only Jeju caves can offer.

• Journal of Conservation Science
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• v.27 no.1
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• pp.101-114
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• 2011

The rock-carved Buddha statue at Dosolam (Korea Treasure No. 1200) of the Seonunsa temple in Gochang is unique style sculptured on natural rock cliff of 13.0m height. The Buddha statue is composed of volcanic complex with tuff, dacitic tuff breccia, tuff bereccia and lithic tuff. Especially, the Buddha statue is characterized by hydrothermal alteration and fragmentation on the upper and lower part. As a result of damage diagnosis, exfoliation and detachment of physical weathering are high of 11.3% and 9.3%, respectively. Infrared thermography analysis, exfoliation and micro-cracks occurred in the measuring parts that have not been confirmed by naked eyes. Chemical index of alteration and weathering potential index of host rock for the Buddha statue are 55.16 to 64.01 and 6.14 to 9.92 which are represented within highly weathering degree. In surface, dark black, reddish brown and white discoloration are observed prominently in the lower. Brown discoloration 6.9% is highest. According to the P-XRF measurements, high concentration of Fe in common, in part of dark black discoloration was Mn, white and brown discoloration in part of S and Ca content were higher. Biological weathering that yellowish brown and dark gray crustoes lichenes appeared by 20.8% and 13.3%, respectively. Therefore, comprehensive deterioration rate of Buddha statue show physical damage by 21.2%, discoloration for inorganic contaminants by 10.8% and biological damage by 39.4%. Ultrasonic velocity measurement carried out of Buddha statue on the surface by 555 points. Measured value of ultrasonic velocity was about 2,273m/s(1,067 to 3,215m/s, and weathering coefficient is 0.5(0.4 to 0.8) that progress on MW(moderately weathered) to HW(highly weathered) grade of rocks.

• Applied Microscopy
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• v.29 no.2
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• pp.211-221
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• 1999

The ultrastructural changes of cuticular surface, excretory and digestive organs of Anisakis simplex larvae chronologically recovered from experimental cats were observed with a SEM and TEM. The larva recovered from an experimental cat at 3 days post-infection (PI) retained the cuticular surface with regular transverse striations and a longitudinal groove on the lateral side of body. This finding suggests that the molting of the 3rd stage larva of A. simplex to 4th one occurred from the 3rd day after infection in cats. The excretory organ (renette cell) consisted of a large cell with numerous ductules ramified from the main duct, mitochondria and secretory granules in cytoplasm. Secretory granules in the renette cell of larvae recovered at 24 hours PI were round whereas those of control and larvae recovered at 6 hours PI were amorphous. Muscular esophagus and ventriculus also retained many secretory granules in the cytoplasm. The secretory granules in these organs of larvae recovered at $6\sim24$ hours PI were electron-dense and widely distributed whereas those of control worm were packed in a pocket and retained various electron densities. In the cytoplasm of intestinal epithelial cells, numerous fine glycogen particles and mitochondria were distributed. The chronological changes of secretory granules in renette cell, muscular esophagus and ventriculus seem to be related with the worm penetration into host tissue.

• Horticultural Science & Technology
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• v.33 no.1
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• pp.70-82
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• 2015

This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

• Journal of agriculture & life science
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• v.45 no.2
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• pp.43-50
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• 2011

Hibiscus cannabinus L. is a plant in the Malvaceae family, that was seeded at June 1st in 2010 and harvested at November 18th. The present study was designated to investigate the safety for host cells, antibacterial effects of Hibiscus cannabinus L. of flower (HCME-F) or leaf (HMEF-L) methanol extract for typical Gram's positive bacteria (St. aureus and Str. epidermidis) or Gram's negative bacteria (S. typhimurium and E. coli). In treatment of different concentrations of HCME-F or HMEF-L (1, 50 and $100{\mu}g/ml$), cytotoxic effects were not shown to RAW 264.7 cells until 24 h incubation. In determination of antibacterial activity of HCME-F or HMEF-L, the antibacterial activities for St. aureus and Str. epidermidis were markedly increased compared to that of untreated control group, but antibacterial activity of HCME-F or HMEF-L for S. typhimurium and E. coli were not changed. Taken together, we demonstrated that methanol extract of HCME-F or HMEF-L showed the safety for RAW 264.7 cells and antibacterial activities for Gram's positive pathogenic bacteria St. aureus and Str. epidermidis. These findings suggest that a methanol extract of Kenaf flower or leaf may be useful alternatives of conventional chemotherapies for dermatitis and mastitis causing Gram's positive pathogens such as Stapylococcus spp. and Streptococcus spp. in domestic animals and humans.