Jeong, Tae Hyug;Youn, Joo Yeon;Ji, Keunho;Seo, Yong Bae;Kim, Young Tae
Journal of Life Science
/
v.24
no.4
/
pp.343-351
/
2014
Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.
Ko, Myoung-Suk;Bae, Kee Hwa;Song, Gwanpil;So, In Sup
Journal of Plant Biotechnology
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v.40
no.2
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pp.79-87
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2013
The aim of this study was to establish plant regeneration from leaf explants of Sedum tosaense Makino, which is globally rare and endangered species. The leaf explants of S. tosaense were cultured on the MS medium supplemented with different concentration of BA and NAA for callus induction. Callus induction was showed the highest (100%) on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. The highest number of shoots were regenerated when callus were cultured on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA for 5 weeks. The axillary bud were cultured on the MS media supplemented with combination of BA and NAA for in vitro propagation. The highest number of adventitious shoot (7.9 per explants) formed at $1.0mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ BA. For rooting, MS medium supplemented with or without $2.0g{\cdot}L^{-1}$ activated charcoal was tested. The optimal results were observed using MS medium supplemented with $2.0g{\cdot}L^{-1}$ activated charcoal, on which 85.7 (No. of root), 4.6 cm (length of root). 1,200 ppm $CO_2$ and 350 ppm $CO_2$ were supplied for make certain the effects of $CO_2$ on pre-acclimatization by photoautotrophic culture. 1,200 ppm $CO_2$ treatment was established higher than 350 ppm $CO_2$ treatment. Soil acclimatization of in vitro plantlets was the best in mixture soil consisted of peat moss and perlite with 100% survival rate and they showed the maximum growth.
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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v.7
no.3
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pp.181-194
/
2002
Parasitism is a one-sided relationship between two organisms in which one benefits at the expense of the other. Parasitic dinoflagellates, particularly species of Amoebophrya, have long been thought to be a potential biological agent for controlling harmful algal bloom(HAB). Amoebophrya infections have been reported for over 40 species representing more than 24 dinoflagellate genera including a few toxic species. Parasitic dinoflagellates Amoebophrya spp. have a relatively simple life cycle consisting of an infective dispersal stage (dinospore), an intracellular growth stage(trophont), and an extracellular reproductive stage(vermiform). Biology of dinospores such as infectivity, survival, and ability to successfully infect host cells differs among dinoflagellate host-parasite systems. There are growing reports that Amoebophrya spp.(previously, collectively known as Amoebophrya ceratii) exhibit the strong host specificity and would be a species complex composed of several host-specific taxa, based on the marked differences in host-parasite biology, cross infection, and molecular genetic data. Dinoflagellates become reproductively incompetent and are eventually killed by the parasite once infected. During the infection cycle of the parasite, the infected host exhibits ecophysiologically different patterns from those of uninfected host in various ways. Photosynthetic performance in autotrophic dinoflagellates can be significantly altered following infection by parasitic dinoflagellate Amoebophrya, with the magnitude of the effects over the infection cycle of the parasite depending on the site of infection. Parasitism by the parasitic dinoflagellate Amoebophrya could have significant impacts on host behavior such as diel vertical migration. Parasitic dinoflagellates may not only stimulate rapid cycling of dissolved organic materials and/or trace metals but also would repackage the relatively large sized host biomass into a number of smaller dinospores, thereby leading to better retention of host's material and energy within the microbial loop. To better understand the roles of parasites in plankton ecology and harmful algal dynamics, further research on a variety of dinoflagellate host-parasite systems is needed.
Kim, Seong-Yong;Son, Sung-Kweon;Kim, Jae-Ryong;Kim, Jung-Hye
Journal of Yeungnam Medical Science
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v.10
no.2
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pp.432-444
/
1993
Multidrug resistance(MDR) phenotype is frequently observed in animal and human cancer cell lines selected for in vitro resistance to a single chemotherapeutic agent. It is characterized by the diminished drug accumulation and is related to the drug efflux mechanism in resistant cells. In the present study, adriamycin resistant cells(L1210-$AdR_6$ : $10^{-6}M$ adriamycin, $-AdR_5$ : $10^{-5}M$) and vincristine resistant cells (L1210-$VcR_7$ : $10^{-7}M$ vincristine, $-VcR_6$ : $10^{-6}M$) were produced from mouse lymphoblastic leukemia cell line L1210. Growth profiles of survived cells were observed for 5 days with MTT(thiazolyl blue) assay and resistance was compared with $IC_{50}$(drug concentration of 50% survival reduction in absorbance). Resistant cells proliferated more slowly than sensitive cell. Doubling times were 29.7hr in L1210, 68.7hr in L1210-$AdR_5$ and 58.2hr in $-VcR_6$. MDRs expressed as resistance factor were as follows, L1210-$AdR_5$ was 76.4 times for vincristine, L1210-$VcR_6$ was 96.4 times for adriamycin. The cell membrane proteins with three different M.W. were recognized to be related resistance, 220, 158, and 88 kd in L1210-$AdR_5$, 158, 140 and 88 kd in L1210-$VcR_6$ by SDS-PAG electrophoresis. Cell surface membrane proteins were identified by radio-iodination and autoradiogram, their molecular weights were 158, 72.8, and 42.4 Kd in L1210-$VcR_6$.
This experiment was conducted to clarify the characteristics of germination and emergence of Panicum dichotomiflorum which is a noxious weed species in direct-seeded rice field. P. dichotomiflorum was planted with several treatments such as different depths of irrigation to verifiy the ecological habits of seedling emergence and growth. In order to know the germination characteristics, Panicum dichotomiflorum seeds, pretreated with low-temperature($4^{\circ}C$) stratification for breaking the dormancy, were germinated under different temperature regimes and water potentials. Germination rates of P. dichotomiflorum was increased from 0% of dormant seed to 1%, 35% and 44% by stratification for 21, 28 and 42 days, respectively. Two dominant weed species in directseeded rice fields, Echinochloa crus-galli and Panicum dichotomitlorum, showed different germination habit under different temperature regimes. Echinochloa crus-galli showed more higher germination rate than Panicum dichotomiflorum at relatively low temperature regime(20/$10^{\circ}C$). Both species germinated faster at 30/$20^{\circ}C$ than at 20/$10^{\circ}C$. When the water potential was lowered, germination of Panicum dichotomiflorum was reduced more drastically than Echinochloa crus-galli. The critical water potential for germination of P. dichotomiflorum was -0.7MPa but Echinochloa crus-galli was affected slightly by the same water potential. The results showed that Echinochloa crus-galli can germinate under more wide range of soil water potential than Panicum dichotomiflorum. Emergence of P. dichotomiflorum was highly affected by irrigation depth and the level of water table. When the depth of irrigation water was increased, emergence of P. dichotomiflorum was dragged and emergence rate showed significant difference under the irrigation depth deeper than 6cm. The maximum depth of irrigation water for survival of Panicum dichotomiflorum seedling was 9cm.
This experiment was conducted to determine the economic application method of Yakto among 6 application method : amount of Yakto application $3\;plot(70\;l,\;25.5\;l,\;13,3\;l/kan;\;180cm{\times}90cm),$ and added side dressing(3 l/kan) each plot. The reduced amoumt of Yakto application(25.5 l, 13.3 l/kan) were lower the contents of inorganic nitrogen, $P_{2}O_{5},\;K,\;Ca$ and Mg, but similar pH in bed soil compared with standard Yakto application(70 l/kan). Side dressing applicated on 29 May increased the contents of inorganic nitrogen, $P_{2}O_{5},\;K,\;Ca,\;Mg$ and salt concentration in bed soil compared with nonside dressing in each Yakto application plot(70 l, 25.5 l, 13,3 l/kan). The reduced amoumt of Yakto application(25.5 l/kan) added side dressing(3 l/kan) were similar the contents of inorganic nitrogen, K, Mg and salt but lower $P_{2}O_{5}$ and Ca in bed soil compared with standard Yakto application(70 l/kan) in June. The reduced amoumt of Yakto application(25,5 l/Kan) added side dressing(3 l/kan) was similar the contents of P, K, Ca and Mg but lower N in leaves compared with standard Yakto application(70 l/kan). The reduced amoumt of Yakto application(25.5 l/kan) added side dressing(3 l/kan) was shorter stem length, and higher $5{\%}$ chlorosis of leaf tip, but similar emergence ratio, survival ratio, leaf senescence ratio, chlorophyll, stem diameter and leaf area compared with standard Yakto application(70 l/kan). These suggested that 25.5 l Yakto application added side dressing(3 l/kan) per kan might be possible economical application method of Yakto.
To study the effect of salinity on maturation and spawning, two series of experiments were carried out during November, 1998- July, 1999 and January, 1999- March 2000. In the first series, the control group (C) was reared at 14.3$^{\circ}C$ and 35.1$\textperthousand$ S, but the experimental group was reared at 33.8$\textperthousand$ S. In the second series, they were also exposed to approximately the same temperature but the experimental group was reared at still lowered salinity of 2p.6$\textperthousand$. Survival, food intake and growth of the experimental groups in either series reared at lower salinities were higher than those of their respective control groups. At lower salinity, larger number of females attained spontaneous sexual maturity and successfully spawned. Hatching success of the eggs spawned by the experimental females reared at lower salinities was also higher (54-56$\textperthousand$) than that (~19%) of the control group reared at higher salinity.
In this study, the radioprotective effects of Grifola umbellata hot water extracts (Gu-extract) on mice were investigated. Single pre-administration of Gu-extract increased the 40-day survival ratio of irradiated mice from 65.5% to 78.6%. The growth of 3 week old male mice in the irradiated group was slightly retarded as compared to those of the control and Gu-extract treated mice. The average spleen and thymus weights of the irradiated mice were lower than those of the control and Gu-extract treated mice. The weight reduction of testis in the irradiated mice was significant. While it was relatively slight in the Gu-extract treated mice as compared to that of control mice. No significant difference in the weight was observed in heart, kidney or liver among three groups. The leukocytes of the Gu-extract treated mice did not decrease dramatically as in the irradiated group, but recovery patterns were similar in both groups. Reduction of erythrocytes were similar in both groups but its recovery occurred more rapidly in the extract treated group. The glucose level of the Gu-extract treated group did not change during the period examined, while it was still higher in the irradiated group than the level in the control group in two weeks. The cholesterol levels in the irradiated and the Gu-extract treated groups were higher than that of control group on day 7, but decreased to the level of the control group on day 14. No difference was observed in total protein amount of the serum among the three groups. SDS-polyacrylamide gel electrophoresis of the soluble proteins extracted from various organs did not reveal differences to any extent in all groups except in the livers of the irradiated and extract treated groups, in which some proteins were missing or less present.
Jun-ki Lee;Se-Hui Lee;Dong Ju Seo;Kang-Hee Lee;Sojung Park;Sun Park;Taekyung Kim;Jin-Young Yang
Journal of Life Science
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v.33
no.2
/
pp.158-168
/
2023
Natural products have been used to mitigate the effects of cancer and infectious diseases, as they feature diverse bioactivities, such as antioxidant, antibacterial, anti-inflammatory, and immunomodulatory effects. Here, we chose 10 natural products that are well-known as pulmonary enhancers and investigated their bactericidal effects on Streptococcus pneumoniae. In the disk diffusion assay, the growth of S. pneumoniae was significantly regulated by G. fructus treatment regardless of extraction method used. We first adopted spraying as a novel delivery method for G. fructus. Interestingly, mice exposed to G. fructus three times a day for 2 weeks were resistant to S. pneumoniae intranasal infection (shown both through body weight loss and survival rates compared to the control group). Moreover, we confirmed that exposure to G. fructus regulated the colonization of the bacteria despite the sustained inflammation in the lung after exposure to S. pneumoniae, indicating that migrated inflammatory immune cells may involve a host defense mechanism against pulmonary infectious diseases. While a similar number of granulocytes (CD11b+Ly6C+Ly6G+), neutrophils (CD11b+Ly6CintLy6G+), and monocytes (CD11b+Ly6CintLy6G-) were found between groups, a significantly increased number of alveolar macrophages (CD11b+CD11chiF4/80+) was detected in BAL fluids of mice pre-exposed to G. fructus at 5 days after S. pneumonia infection. Taken together, our data suggest that this usage of G. fructus can induce protective immunity against bacterial infection, indicating that facial spray may be helpful in enhancing the defense mechanism against pulmonary inflammation and in evaluating the efficacy of natural products as immune enhancers against respiratory diseases.
This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.
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