• Journal of Microbiology
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• v.37 no.3
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• pp.141-147
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• 1999

Thirteen transcriptionally-fused carbon starvation mutants, derived from Pseudomonas putida ATCC 12633, were analyzed for their survivability and transcriptional induction profiles upon carbon starvation. One of these mutants, MK114, which exhibited the lowest survivability and the highest induction rate, was selected and further examined under different starvation (nitrogen and phosphate) and stress (osmolarity, H2O2, salts, alcohol, and heat) conditions. Under all tested conditions MK114 induced ${\beta}$-galactosidase activity, implying that the interrupted gene (cst114) is a general starvation and stress response gene. The rate of induction ranged from 2.6-fold for phosphate starvation to 3.7-fold for osmotic shock. The mini-Tn5 flanking DNA was cloned from the chromosome of MK114. The cloned DNA fragment exhibited carbon starvation activity, indicating that this fragment contains a carbon starvation-related promoter region. This region was partially sequenced. Possible physiological roles of Cst114 in a carbon sensing mechanism and in other stress responses are also discussed.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.326-328
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• 2001

Pseudomonas diminuta (ATCC 19146) has been typically used in the bacterial challenge test for validation of the sterilizing filtration process. Cell size is critical for determining the retention characteristics of membrane filters with pore-size of $0.2{\mu}m$. The changes of cell sizes after osmotic shocks at 150, 260, 500, and 700 mosM were measured by a particle size analyzer and the changes of their buoyant densities were analyzed with a Percoll gradient. The results indicated that there were no significant differences when cells were cultured in 260 mosM medium and osmotically shocked at 500 and 700 mosM. However, the osmotically shocked cells at 150 mosM showed a 38% increase of the cell size compared to the cells at 260 mosM. From these study, we concluded that the worst case condition for validation of a sterilizing filter would be 500 mosM, not because of changes in the cell size, but due to decrease in cell viability under those conditions.

• Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.209-218
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• 2009

We examined physiological and histological responses related to the survival, oxygen consumption, excretion, and O/N ratio of the ark shell, Scapharca subcrenata, as a result of salinity changes. The 20-day $LS_{50}$ (median lethal salinity) at $15^{\circ}C$ was 13.87 practical salinity units (psu; confidence limits 10.30-18.74 psu), whereas the 14-day $LS_{50}$ at $25^{\circ}C$ was 12.59 psu (confidence limits 8.03-18.16 psu). In conditions of decreasing salinity, the osmolarity of individuals acclimated within 5 h above 26.4 psu but required more than 60 h below 13.2 psu. Oxygen consumption and ammonia excretion rates varied irregularly as salinity decreased. The O/N ratio was 19 and 27 at water temperatures of $15^{\circ}C$ and $25^{\circ}C$, respectively, but decreased to 1-10 as salinity declined. The effects of decreasing salinity were observed in the histological changes to each organ of S. subcrenata. As salinity decreased, cilia fell off, the epithelial layer underwent necrosis and vacuolation, the connective tissue layers of the mantle and visceral mass were destroyed, and hemocytes increased in the gills. The results of this study could prove important in investigating causes of mass mortality and managing shellfish aquaculture farms.

• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1377-1381
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• 2014

In order to discover and develop novel signaling inhibitors from plants, a screening system was established targeting the two-component system of Cryptococcus neoformans by using the wild type and a calcineurin mutant of C. neoformans, based on the counter-regulatory action of high-osmolarity glycerol (Hog1) mitogen-activated protein kinase and the calcineurin pathways in C. neoformans. Among 10,000 plant extracts, that from Harrisonia abyssinica Oliv. exhibited the most potent inhibitory activity against C. neoformans var. grubii H99 with fludioxonil. Bioassay-guided fractionation was used to isolate two bioactive compounds from H. abyssinica, and these compounds were identified as chebulagic acid and chebulanin using spectroscopic methods. These compounds specifically inhibited the calcineurin pathway in C. neoformans. Moreover, they exhibited potent antifungal activities against various human pathogenic fungi with minimum inhibitory concentrations ranging from 0.25 to over $64{\mu}g/ml$.

• YAKHAK HOEJI
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• v.56 no.2
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• pp.74-79
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• 2012

Citrate-capped silver nanoparticles (AgNPs) are widely used in industry, consumer products and medical appliances. However, information on the toxicity, environmental fate and toxicokinetics are not enough. In this study, stability of citrate-capped AgNPs was investigated using different types of isotonic solution, which is important in the toxicokinetic study by the exposure route of intravenous injection. Size, morphology, zeta potential and ion formation were investigated in isotonic solutions for the physico-chemical characterization of AgNPs. Aggregation and precipitation of AgNPs were observed in saline or phosphate-buffered saline while they were stable without precipitation in 2% glycerol of isotonic solution. The average size of AgNPs in 2% glycerol was 6~10 nm, which was almost same as that in water-based suspension of AgNPs. Zeta potential was ranged from -30 mV to -60 mV, which was in the range of original stock AgNPs. The stability was maintained during the whole experimental period of 48 hours. Furthermore, the stability was not changed in different temperature (10~36$^{\circ}C$) and at different concentrations (10~1,000 ppm). The osmolarity of the AgNPs suspension was $299{\pm}1$ mOsm/kg which was in isotonic range. These data suggest that AgNPs in 2% glycerol solution can be used for the preparations of intravenous injection for toxicokinetic study without undesired disturbance of blood isotonicity.

• Mycobiology
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• v.42 no.2
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• pp.152-157
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• 2014

The iron uptake and utilization pathways play a critical role in allowing human pathogens, including Cryptococcus neoformans, the causative agent of fatal meningoencephalitis, to survive within the mammalian body by competing with the host for iron. Here we show that the iron regulon is also required for diverse environmental stress responses and that in C. neoformans, it is regulated by the high-osmolarity glycerol response (HOG) pathway. Between CFO1 and CFO2, two ferroxidase genes in the iron regulon, CFO1 but not CFO2 was induced during oxidative and osmotic stress. Interestingly, we found that the HOG pathway repressed basal expression of both CFO1 and CFO2. Furthermore, when the HOG pathway was blocked, CFO2 also responded to oxidative and osmotic stress and the response of CFO1 was increased. We also established that CFO1 plays a major role in responding and adapting to diverse environmental stresses, including oxidative and genotoxic damage, osmotic fluctuations, heavy metal stress, and stress induced by cell membrane destabilizers. Therefore, our findings indicate that in C. neoformans, the iron uptake and utilization pathways are not only required for iron acquisition and survival, but also play a significant role in the environmental stress response through crosstalk with the HOG pathway.

• Toxicological Research
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• v.11 no.2
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• pp.199-203
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• 1995

The focus of this study was to investigate that cellular parameters and glucose uptake might be altered by extracellular calcium and starvation. Addition of 1 mM $Ca^{++}$ to hepatocytes (equalling to the free calcium concentration of blood) significantly increased intracellular $Na^+$ and decreased $Na^+$ & LDH leakage. This pertains to the hepatocytes of control rats as well as those of rats fasted for 24 and 48. hr. These effects might be come from the membrane-stabilizing effects of calcium. But calcium had no effects on cell volumes, superoxide-formation and glucose uptake. Actually hepatocytes of starved rats showed changes in several cellular parameters. Starvation increased LDH leakage, glucose uptake and the total concentration of $Na^+$ and $Na^+$ whereas it markedly decreased cell volumes. Since total tonicity remained unchanged, intracellular $Na^+$ and $Na^+$ could contribute to a higher share of total osmolarity in starvation. Starvation increased the cytoplasmic pH because $R-NH^{3+}$ions and their corresponding counterions disappeared. This increase may be related to suppress the protonization of amino groups in proteins. Starvation decreased hepatic glycogen, a major compound that affects cytosolic volume of hepatocytes. The data indicate that starvation increases the glucose transport activity. The possible molecular basis will be discussed.

• Journal of Nutrition and Health
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• v.37 no.5
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• pp.364-372
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• 2004

The purpose of this study was to investigate the possibility of medicinal plants application as an edible functional food resource. We carried out to develop a traditional functional beverage by using hot-water extraction of 4 medicinal plants (Polygonatun sibiricum, Ophiopogonis radix, Lycii fructus, Schizandriae frutcus) and we examined the effects of drink on physiological response during exercise and recovery phase. The subjects were male baseball players, and exercise protocol was performed with 45 minutes treadmill running and 85％ VO$_2$ max intensity. Brix, pH and titratable acidity of developed drink were 9.5％, 3.3 and 0.22％. The approximate nutritional composition of beverage was carbohydrate, 5.98％, total dietary fiber, 0.42％, crude protein, 0.70％, crude fat, 0.20％ and crude ash, 0.20％. Developed. traditional functional beverage contained K (4.00 mg％), Na (3.68mg％), Ca (2.54mg％), Mg (1.60mg％) and Fe (0.29mg％). Developed beverage drinking group showed a lowest heart rate during exercise and recovery phase than the other two group (water group and ion beverage group). In the change of blood lactate concentration, developed beverage intake group showed a significant lowest values during exercise and recovery phase. And blood hematocrit values and osmolarity were lowest in the developed beverage intake group. In these results, exercise performance capacity was expected to improve most highly with the developed drink from composition with medicinal plants. Also developed beverage was effective in recovery of exercise-induced fatigue. Thus developed drink with medicinal plants can be used as a functional material improving decrease fatigue effects in beverage industry.

• KSBB Journal
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• v.8 no.2
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• pp.97-103
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• 1993

This study was investigated as a step for the purpose of successful introduction of cytoplasmic inherited characters between the different plants. Cytoplasts were separated from the protoplasts of CMS(cytoplasmic male sterility) line such as MS Burley 21 which carried from Nicotiana megalosiphon. The cytoplasts were fused to protoplasts derived from Nicotiana tabacum Br 64 with PEG(polyethylene g1yco1). The cytoplasts were separated by density gradient centrifugation. Efficient separation of cytoplasts depended on the difference of specific density of gradient solution. However, the iso-osmolality of gradient solution was not important to separate the cytoplasts. The cells for a cybrid were fused with 50% concentration of PEG.

• Mycobiology
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• v.37 no.3
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• pp.161-170
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• 2009

Cryptococcus neoformans is a basidiomycete human fungal pathogen that causes meningoencephalitis in both immunocompromised and immunocompetent individuals. The ability to sense and respond to diverse extracellular signals is essential for the pathogen to infect and cause disease in the host. Four major stress-activated signaling (SAS) pathways have been characterized in C. neoformans, including the HOG (high osmolarity glycerol response), PKC/Mpk1 MAPK (mitogen-activated protein kinase), calcium-dependent calcineurin, and RAS signaling pathways. The HOG pathway in C. neoformans not only controls responses to diverse environmental stresses, including osmotic shock, UV irradiation, oxidative stress, heavy metal stress, antifungal drugs, toxic metabolites, and high temperature, but also regulates ergosterol biosynthesis. The PKC(protein kinase C)/Mpk1 pathway in C. neoformans is involved in a variety of stress responses, including osmotic, oxidative, and nitrosative stresses and breaches of cell wall integrity. The $Ca^{2+}$/calmodulin- and Ras-signaling pathways also play critical roles in adaptation to certain environmental stresses, such as high temperature and sexual differentiation. Perturbation of the SAS pathways not only impairs the ability of C. neoformans to resist a variety of environmental stresses during host infection, but also affects production of virulence factors, such as capsule and melanin. A drug(s) capable of targeting signaling components of the SAS pathway will be effective for treatment of cryptococcosis.