Objective: Previously, we identified differentially expressed genes between GV and MII stage mouse oocytes using ACP technology. When we study one of GV selective genes, Obox family, we found Obox4 mRNA expression in ovaries that has been reported as expressed exclusively in testis. Therefore, this study was conducted for characterization and functional analysis for Obox4. Methods: Expression of Obox4 mRNA was examined in gonads and oocytes by RT-PCR. To determine the role of Obox4 in oocyte maturation, Obox4 dsRNA was microinjected into the cytoplasm of GV oocytes followed by 16 h of incubation in the plain medium or by 24 h of incubation in the medium containing IBMX. After RNAi, phenotypes and maturation rates were observed, change in mRNA expression was evaluated, and chromosomal status was confirmed by orcein staining. Results: Obox4 has minimal expression in the ovary compared to that of the other family members. When oocytes were cultured for 16 h in M16 medium after RNAi, maturation rate was not changed significantly, compared with that of non-injected or buffer-injected control oocytes. Surprisingly, however, when oocytes were cultured for 24 h in M16 containing IBMX, in which oocytes were supposed to arrest at GV stage, Obox4 RNAi oocytes were advanced to MI and MII. Spindle structure was disappeared and the chromosomes were condensed in the oocytes after Obox4 RNAi. Conclusions: This is the first report on the expression of Obox4 in the ovary and oocytes. Results of the study suggest that Obox4 plays a crucial role in spindle formation and chromosome segregation during meiosis in oocytes. In addition, Obox4 may play an important role in cAMP-dependent signal cascades of GV-arrest in mouse oocytes.
Several medicinal herbs, which are nontoxic and have been used widely in traditional folk medicine, were extracted and antimicrobial activity of the extracts was investigated against various foodborne pathogens or food poisioning microorganisms. The ethanol extract of Sansa, Hwangryun, Cheukbaek, and Seokchangpo showed strong antimicrobial activities against Gram postive and Gram negative bacteria, whereas those of Sakunja, Sukjihwang and Baekji had little antimicrobial activities on microorganisms tested. Among medicinal herb extracts, ethanol extract of Coptis chinensis Franch (hwangryun) showed the strongest antimicrobial activity. Antimicrobial activity of ethanol extract of Coptis chinensis Franch was not destroyed by heating at $100^{\circ}C$ for 60 min and at $121^{\circ}C$ for 30 min, which is very stable over heat. The pH effect on minimal inhibitory concentrations (MIC) of the extract of Coptis chinensis Franch indicated that MIC was reduced with increasing the pH value of the medium. The inhibitory effect of partially purified substance from the ethanol extract of Coptis chinensis Franch on the growth of Listeria monocytogenes and Staphylococcus aureus was investigated. Growth of those strains occurred at the concentration of $100{\;}{\mu}g/mL$ and were inhibited at $500{\;}{\mu}g/mL$, whereas those strains was completely inactivated in the presence of $1000{\;}{\mu}g/mL$.
The nutritive value and the effect of tannins on the utilization of foliage from three commonly used legumes, Acacia nilotica, Albizia procera, and Sesbania acculeata, were determined. Three mature rumen-fistulated bullocks were used to study in sacco degradability and twelve adult sheep were randomly allocated on the basis of live weight to 4 groups of 3 in each to study the in vivo digestibility of the foliages. In all foliages, the contents of crude protein (17 to 24% of DM) were high. Fibre was especially high in Albizia (NDF 58.8% of DM vs. 21% in Sesbania and 15.4% in Acacia). Contents of both hydrolysable (4.4 to 0.05%) and condensed tannins (1.2 to 0.04%) varied from medium to low in the foliages. Acacia contained the highest level of total phenolics (20.1%), protein precipitable phenolics (13.2%) and had the highest capacity to precipitate protein (14.7%). Drying in shade reduced the tannin content in Acacia and Albizia by 48.6 and 69.3% respectively. The foliages ranked similarly for each of the different methods used to estimate tannin content and activity. Acacia and Sesbania foliage was highly degradable (85-87% potential degradability of DM in sacco), compared to Albizia (52%), indicating a minimal effect of tannins in Acacia and Sesbania. Yet, in vitro, the tannins in the Acacia inhibited microbial activity more than those in Albizia and Sesbania. Following the addition of polyethylene glycol to neutralise the tannins, gas production and microbial growth increased by 59% and 0.09 mg RNA equiv./dg microbial yield respectively in the Acacia, compared to 16-17% and 0.06 mg RNA equiv./dg microbial yield in the other foliages. There was a trend for low in vivo apparent digestibility of N in the Acacia (43.2%) and Albizia (44.2%) compared to the Sesbania (54.5%) supplemented groups. This was likely to be due to presence of tannins. Consistent with this was the low N retention (0.22 and 0.19 g N/g NI) in sheep supplemented with Acacia and Albizia compared to that for the Sesbania (0.32). Similarly, a trend for poor microbial N yield was observed in sheep fed these foliages. Across the foliages tested, an increase in tannin content was associated with a reduction in ruminal fermentation, N digestibility and N retention. For overall nutritive value, Sesbania proved to be the superior forage of the three tested.
The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.
Song, Ji Hyun;Lee, Jin Hwa;Kim, Eun Jin;Kim, Yun Hee
The Journal of Pediatrics of Korean Medicine
/
v.32
no.3
/
pp.1-15
/
2018
Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.
Vanadium is an essential trace element but has not been identified with a specific biogical role. To study the direct effects of vanadium on osteoblast, we incubated murin osteoblast-like (MC3T3-El) cells with various corcentration of vanadium oxide & sodium orthovanadate. This study was designed to investigate the effect of vanadium on DNA synthesis, alkaline phosphatase (ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in murin osteoblast-like (MC3T3-El) cells. The cells were cultured in $\alpha-minimal$ essential medium$(\alpha-MEM)$ supplemented with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concenoation of vanadium oxide & sodium orthovanadate. Quiescent cultured MC3T3-El cells incubated for 24 hours with 2,5,10,15,20 ${\mu}M$ vanadium oxide incorporated $[^3H]Thymidine;$ every concentration showed increases in $[^3H]Thymidine$ incorporations dose dependant manner, the greatest response occurred at $20{\mu}M$. Quiescent cultured MC3T3-E1 cells incubated for 3days with 2,5,10,15,20 ${\mu}M$ vanadium oxide, for 2days with sodium orthovanadate and alkaline phosphatase was assayed with disodium phenyl phosphate as substrate. Vanadium oxide increased the alkaline phosphatase content in MC3T3-El cells at $2{\mu}M\;&\;6{\mu}M$ ; the greatest response occurred at $2{\mu}M$. But decreased at other content sodium orthovanadate increased alkaline phosphatase content in MC3T3-El cells at all concenoation ; the greatest response occurred at $4{\mu}M$. Quiescent cultured MC3T3-El cells incubated for 3days with $5,10{\mu}M$ vanadium oxide , with $5,8{\mu}M$ sodium orthovanadate and cAMP formation was measured by Radioimmunoassay(RIA). Vanadium oxide & sodium orthovanadate showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-El cells. Quiescent cultured MC3T3-El cells incubated for 24hours with $10,20{\mu}M$ vanadium oxide, with $5,10{\mu}M$ sodium orthovanadate and Type I $\alpha$ 2 collagen ribonucleic acid (mRNA) expression was studied by Nothern blot analysis. Northern blot analysis of vanadium oxide treated cells showed decreasing effects 0& sodium orthovanadate revealed increasing effects in type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level.
Proceedings of the Korea Society for Industrial Systems Conference
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1999.12a
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pp.121-129
/
1999
When continuous media are transmitted over the communication networks, asynchrony which can not maintain temporal relationships among packets may occur due to a random transit delay. There exist two types of synchronization schemes ; for guaranteed or non-guaranteed resource networks. The former which applies a resource reservation technique maintains delay characteristics, however, the latter supply a best-effort service. In this paper, I propose a intra-media synchronization scheme to transmit continuous media on general networks not guaranteeing a bounded delay tome. The scheme controls transmission times of the packets by estimating next delay time with the delay distribution. So, the arriving packets may be maintained within a limited delay boundary, and playout will be performed after buffering to smoothen small delay variations. The continually increasing delay due to network overload causes buffer underflow at the receiver. To solve it, the transmitter is required to speed up instantaneously. Too much increase of transmission-rate may cause network congestion. At that time, the transmitter drops the current packet when informed excessive delay from the receiver.
Lattice steel towers for overhead transmission lines have been replaced by tubular steel poles due to the visual impact of large and complex shape of truss type. Demand for tubular steel poles consisting of a single frame member continues to grow because of its advantages such as visual minimization, architectural appeal and minimal site consumptions. However, there are some constraints on the transportation and construction. As the diameter of tower base has been enlarged, it may exceed minimum height limit required to pass the tunnel in case of land transportation. Also, in a narrow place where it is not easy to secure the installation areas such as mountainous places, there might be some areas wherein it must secure a wide working space so that large vehicles and working cranes will be allowed to enter. In this paper, we presented a vertical separated tubular steel pole, which is a new type of support that can be implemented for general purpose such as mountainous areas or narrow areas to improve the issues raised by breaking away from the conventional design and fabrication methods. Technical approaches for overcoming the limit of the cross-sectional size is to separate and modularize the cross-section of the tubular steel pole designed with a size that cannot be carried or assembled, and to lighten it with a weight capable of being transported and assembled in a narrow space or mountainous area. As a result of this research, it will be possible to enter small and medium sized vehicles in locations where it is restricted to transport by large-sized vehicles. In the case of mountainous areas, it will be possible to divide it into a weight capable of being carried by a helicopter and it will be easy to adjust and fabricate it with individual modules. Furthermore, in order to break away from the traditional construction method, we proposed the equipment that can be applied to the assembly of Tubular Steel Pole without using a large crane in locations where there is no accessible road or in locations wherein large cranes cannot enter. In particular, this paper shows the movable assembling equipment and some methods that are specialized for vertical separated tubular steel pole consisting of members with reduced weight. The proposed assembly equipment is a device for assembling the body of the Tubular Steel Poles. It will be installed inside the support and the modules can be lifted by using the support itself.
Biological Treatment of Wastewater Containing Chlorinated Phenols by a Mixed Culture. Lee, Wan-Seok1, Sang-Wook Jung, Chan-Sun Park, Byung-Dae Yoon, Jang-Eok Kim\ and Hee-Mock Oh*. Environmental Bioresources Laboratory, Korea Research Institute of Biosicence and Biotechnology, Taejon, Korea, 1 Department of Agricultural Chemistry, Kyungpool< National University, Taegu, Korea - The biodegradation of chlorinated phenols in an artificial wastewater was investigated using a mixed culture. The mixed culture was composed of 8 microorganisms isolated from the soil contaminated with various chlorinated phenols. Pseudomonas sp. BM as a main constituent of a mixed culture was Gram-negative, catalase- and oxidase-positive, and rod-shaped, and did not grow at 41°C. It degraded 99% of initial 500 mg!1 of pentachlorophenol (PCP) in the minimal salts medium as a sole source of carbon and energy within 3 days. The degradation efficiency of Pseu.domon.as sp. BM was not affected by the other organic carbon and nitrogen compounds. Pseudomonas sp. BM was able to grow in a broad range of pH 5 - 8, and degrade 2,000 mg/1 PCP. In the experiment with an artificial wastewater containing chlorinated phenols, the degradation efficiency of the mixed culture was the range of 73% (2,4-dichlorophenol) -96% (2-chlorophenol) during an incubation of 7 days. In a continuous culture experiment, the degradation efficiency of mixed culture plus activated sludge was about 2 times higher than that of the control containing only activated sludge. These results indicate that it is possible to apply the mixed culture to other wastewaters containing chlorinated phenols. Key words: Biodegradation, chlorinated phenols, pentachlorophenol, Pseudomonas sp. BM
The susceptibilities of 45 clinical isolates of bacteroides frogilis to cefaclor, ciproflxacin and imipenem were determined by new method, E-test (AB Bidisk, Solna, Sweden) and were compared with those from conventional agar dilution method by using brain heart infusion, Mueller-Hinton and Wilkins Chalgren agar plates. And the susceptibility of 60 clinical isolates of Bacteroides fragilis group (B. fragilis 45 strains, B. distasonis 6 strains, B. ovatus 5 strains, B. thetaiotaomicron 4 strains) to 5 quinolones (ciprofloxacin, enoxacin, norfloxacin, ofloxacin, pefloxacin) were determined by in vitro agar dilution method. Compared with agar dilution MICs for B. fragilis 45 strains, 90.3% of E-test MICs were within ${\pm}$1 dilution of the agar dilutions, and 98.4% were within 2 dilutions. And there were little effect of different medium bases to determine MICs except Mueller-Hinton agar. On Mueller-Hinton agar, B. fragilis showed have or no growth activity. In vitro susceptibility of B. fragjlis group to quinolones, most of the test strains showed resistant patterns to quinolones except ofloxacin and there was little difference of susceptibility patterns between species of B. fragilis group.
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