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.
This paper attempts to identify the principle of family succession and family patterns of yangban in the late Chosun period through an analysis of male adaptation cases found in family registration records. The primary source of analysis is the family registration documents of Uiseong Kim's from the late 17th century to the early 20th century. As a result, it is found that there is a substantial change in the patterns of family from the early and mid Chosun period to the late Chosun period. The change is the strengthening of the principle of patriarchy succession through male adoption. Looking at the data as a whole, the average number of household members is increased and the membership of kinship also expanded. In contrast to the family patterns of the early Chosun period, not only the patterns of Uiseong Kim's family are predominately immediate family or collateral family but also the majority is extended family in the 18th and 19th centuries. The male adoption cases recorded in Uiseong Kim's family registration documents take up 33.8% of the male adoption cases in the entire family registration documents. This goes to show that the strengthening of the principle of primogeniture succession at a time when child mortality rate is very high resulted in the increase of male adoption. In conclusion, the late Chosun society was a society where the seat of primogeniture was much more important than immediate hereditary members in the family succession.
The Acorn (Quercus acutissima CARRUTHERS), which contains a large quantity of tannin, should be developed as a processed food as the acorn is rich in natural antioxidants and other valuable components. Accordingly, acorn extraction conditions for polyphenol and gallic acid (both antioxidants) were investigated by response surface methodology. The content of polyphenols were determined under 16 different extraction conditions based upon a central composite design. The parameters varied over $30-70^{\circ}C$ of extraction temperature, 1-5 h of extraction time, and 5-25 mL/g of solvent ratio, Gallic acid extraction was optimal at $60-100^{\circ}C$ extraction temperature, 1-5 h of extraction time, and 5-25 mL/g of solvent ratio, Epicatechin content was highest at $56.77^{\circ}C$, 4.16 hand 22.38 mL/g. Catechin content was highest at $52.37^{\circ}C$, 2h and 23.59 mL/g. The maximum catechin content was $91.30{\mu}g/mL$. Epigallocatechin content was influenced by extraction temperature and time. The maximum epigallocatechin content was $1,066.56{\mu}g/mL$ at $61.42^{\circ}C$, 4.17h, and 9.25 mL/g. The maximum value of epicatechingallate content was $125.39{\mu}g/mL$ at $47.72^{\circ}C$, 3.04h, and 24.93mL/g. Epigallocatechingallate content was influenced principally by solvent ratio and the maximum content was $61.38{\mu}g/mL$ at $48.11^{\circ}C$, 2.96h, and 24.95mL/g. The total polyphenol content was maximal at $1,332.75{\mu}g/mL$, after extraction at $61.50^{\circ}C$, 4.24h, at 9.71mL/g. The higher the extraction temperature and the longer the extraction time, the greater the polyphenol content. Gallic acid content was highest, the maximal level was $30.51{\mu}g/mL$ after $65.84^{\circ}C$, 1.65h at 17.17 mL/g, and this was influenced principally by extraction time and solvent ratio.
Anti-obese effects of mulberry (Morus alba L.) root bark was investigated in vitro by measuring its inhibitory effect against 3T3-L1 preadipocyte differentiation and digestive enzymes such as ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase. Ethanol extract of mulberry root bark (MRE) showed the potent inhibitory activities on ${\alpha}$-amylase, ${\alpha}$-glucosidase and pancreatic lipase with $IC_{50}$ values of $7.86{\pm}0.36$, $0.12{\pm}0.03$ and $7.93{\pm}0.11mg/mL$, respectively. Furthermore, MRE significantly suppressed cellular lipid accumulation in 3T3-L1 cells in a dose-dependent manner. To elucidate the mechanism of MRE, we performed qRT-PCR and Western blotting for the expression of genes related with adipogenesis and lipogenesis. Treatment of MRE markedly suppressed the protein expression of $PPAR{\gamma}$, $C/EBP{\alpha}$ and SREBP-1c, as well as FAS and ACC, which are the key transcription factors and metabolic enzymes in adipogenesis and lipogenesis. In addition, qRT-PCR analysis indicated that the anti-adipogenesis effect of MRE might be due to its inhibition at transcription levels. These results demonstrate that MRE can effectively suppress adipocyte differentiation and inhibit key enzymes related to obesity. Our findings suggest that mulberry root bark may have a potential benefit in preventing obesity.
Purpose: The main target of 5-fluorouracil (5-FU) is thymidylate synthase (TS). A high TS expression has been identified as promoting resistance to 5-FU. For colorectal cancers, the response to 5-FU based adjuvant chemotherapy is different according to the microsatellite instability (MSI) status. The reports on the relationship between MSI and the TS expression in colorectal cancer have been inconsistent. No data is available for gastric cancer regarding the relationship between MSI and the TS expression. Therefore, we studied the relationship between MSI and the TS expression in gastric cancer. Materials and Methods: Ninety-nine consecutive patients who underwent radical gastrectomy for gastric cancer from January 2004 to May 2006 at Bundang CHA hospital were studied. MSI was assessed for five markers (BAT25, BAT26, D2S123, D5S346, and D17S250) by PCR and with using an ABI prism 3100 Genetic analyzer. The TS expression was analyzed by immunohistochemistry with using mouse anti-thymidylate synthase monoclonal antibody to the TS 106 clone. Results: Out of the ninety-nine patients, MSS/MSI-L was detected in 92 (92.1%) cases and 7 cases (7.1%) were MSI-H. A negative TS expression was found in 46 (46.5%) cases, a low TS expression was found in 33 (33.3%) and a high TS expression was found in 20 (20.2%). Out of 92 MSS/MSI-L patients, the number of patients with negative, low and high TS expressions were 46 (50%), 30 (32.6%) and 16 (17.4%), respectively. Out of the 7 MSI-H patients, the number of patients with negative, low and high TS expressions were 0 (0%), 3 (42.9%) and 4 (57.1%), respectively. The relationship between MSI-H and a high TS expression was statistically significant. Conclusion: Gastric cancer with MSI-H showed higher levels of a TS expression compared to the gastric cancer with MSS/MSI-L.
its digests on the performance of broiler chicks and taurine content in broiler meat. A total of 100 broiler chickens were assigned to five dietary treatments; Control, PM diet(FM), NaOH treated FM diet(NaOH-FM), HN03 treated FM diet(HN03 - FM) and 0.5% synthetic taurine supplemented diet(Taurine). Treated diets were supplemented with FM or FM digests at the level of 5% to the control diet Treated diets were fed during the last 3wks of 6wks feeding. During the finisher period, weight gain of chicks fed FM treatments tended to decrease in general. Feed intakes of FM and Taurine were significantly lower than the control but were not significantly different from NaOH - FM or HN03- PM. Feed conversion rate(feed intake/gain) of NaOH-FM was significantly higher than those of FM and HNOJ - FM but not different from those of the control and Taurine. Taurine content of the control was highest in heart muscle(1482 J1.g/g) followed by leg muscle(778f.lg/g) and breast muscle(79 J1.g/g). Taurine contents of leg and breast muscle were significantly(P< 0.01) affected by treatments but that of heart muscle was not. Taurine content of Taurine treatments was highest in both leg and breast muscle. Taurine content of leg muscle was increased 170"10 by Taurine supplementation, 123% by FM supplementation, 122% by NaOH - FM supplementation and 63% by HNOJ- FM supplementation compared with control. Taurine content of breast muscle of Taurine treatment was 246% higher than the control, but other treatments were not different from the control. In conclusion, supplementation of feather meal at 5% level of broiler diet can enrich taurine content of leg muscle. Chemical treatments of FM were not effective in improving taurine enrichment of broiler meat.
Nitrogen fertilizers such as urea are readily hydrolyzed in soils to produce ammonium ions which pass through nitrification and denitrification processes. These serial processes have drawn attention due to nitrogen losses, eutrophication, blue baby syndrome, and ozone depletion problems. The purpose of this study was to test the inhibitory effects of hot-water extract and organic solvent fractions of Artemisia asiatica leaves on soil urea hydrolysis and nitrification. In addition, the effects of organic solvent fractions on urease activity and ureolytic bacterial population were also investigated. First, hot-water extract of Artemisia asiatica leaves inhibited soil nitrification substantially with a marginal stimulatory effect on soil urea hydrolysis. Soils treated with hot-water extract of Artemisia asiatica leaves showed significant decreases in the accumulation of soil $NO_3-N$ (~68% decrease) compared with the control soil without the treatment of hot-water extract. In contrast, $CHCl_3$/MeOH fraction and basic aqueous layer of Artemisia asiatica leaves inhibited soil urea hydrolysis very strongly, causing 5.8 and 4.3-fold higher accumulation in amounts of remaining urea-N compared with the non-treated soil. Meanwhile, non of the organic solvent fractions showed any significant effects on soil nitrification inhibition. The inhibition of ureolytic bacterial activity by $CHCl_3$/MeOH fraction and aqueous basic layer of Artemisia asiatica leaves without any effects on urease activity itself led us to conclude that the inhibitions of soil urea hydrolysis were caused by the antagonistic effects on ureolytic bacterial activity.
Park, Mehea;Seo, Jeongmin;Won, Heeyeon;Seo, Jongbun;Moon, Doogyung;Kim, Wooil;Shim, Sangyoun
Horticultural Science & Technology
/
v.33
no.5
/
pp.687-696
/
2015
Black mini tomato 'Hei-G' fruits were harvested at different stages of maturity (immature-mature green and mature-black red) and stored at different temperatures (8, 12, and $20^{\circ}C$) to investigate the quality and lycopene content during storage. Weight loss increased dramatically at higher temperature for both harvesting stages without significant differences. Firmness of immature fruits decreased below the initial level of mature fruit (8.1N) after 5, 8, and 19 days storage, when they were stored at 20, 12, and $8^{\circ}C$, respectively. Soluble solid contents of mature fruit increased at initial storage, and were higher as compared to immature fruits before deterioration at each storage temperature. Decrease in titratable acid of mature fruits depended on storage time and temperature. However, titratable acid of immature fruits showed little change during storage, and so it did not affect flavor. Hunter a value changed greatly in immature fruit stored at high temperature. Unlike ripe tomatoes, there was no significant difference in black tomato Hunter b values of immature and mature fruit at initial and 12 days storage. However, immature fruits stored at $8^{\circ}C$ did not reach full maturity and color development and ripening. High storage temperature increased lycopene production while low storage temperature blocked lycopene development. Shelf life of the immature fruits, which was evaluated by elapsed days to conventional mature stage, was 12 and 15 days when they were stored at 20 and $12^{\circ}C$, respectively. The optimum storage temperature to maintain the quality and lycopene content of mature fruits was $12^{\circ}C$. Moreover, the shelf life of mature fruits stored at $20^{\circ}C$ could reach up to 5 days.
Ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin in chicken muscle were seperated by liquid extraction and determined with high performance liquid chromatography (HPLC) with fluorescence detector. Analysis was carried out using following conditions; Cl8 column (250${\times}$4.6 mm i.d. 5 ${\mu}{\textrm}{m}$ particle size), mobile phase composed of D.W. (containing 0.4% triethylamine and phospholic acid): methanol : acetonitrile (800:100:100, v/v/v), isocratic pump at a flow rate of 1.0 $m\ell$/min and 50 ${mu}ell$ of injection volume, fluorescence detector with EX278 nm/EM.456 nm. The calibration curves of four fluoroquinolones showed linearity (${\gamma}$$^2$$\geq$0.999) at concenration range of 0.025-0.6 $\mu\textrm{g}$/ml. The recoveries in fortified chicken muscle represented more than 80% with low coefficient of variation (〈10%) for concentration range of four fluoroquinolones. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin were 23.5, 3.4, 3.0 and 2.5 ng/g in chicken muscle, respectively. We also monitored fluoroquinolones residue in muscle of chickens (broiler 1:227, Korean native chicken 219, laying chicken 77) using EEC-4-plate screening and HPLC conformation methods. Ten(broiler 5, Korean native chicken 5) out of the fifteen samples which were positively detected by EEC-plate screening method from 1,523 chicken meat were confirmed with ciprofloxacin and enrofloxacin by HPLC. The ranges of residual concentration were 0-0.12 ppm for ciprofloxacin and 0.01-6.79 ppm for enrofloxacin. In conclusion, our method could be applied effectively to determine four fluoroquinolones residues in chicken meat, and further survey for fluoroquinolones residue in chicken meat are needed for more effective control of fluoroquinolones used in livestock.
Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.
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