Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.
Background: We present here the early and midterm surgical results for infective endocarditis and we especially focus on the effect of aggressive reconstruction or root implantation after wide debridement. Material and Method: Between January 1995 and Jun 2006, we enrolled 79 adult infective endocarditis patients who underwent surgical treatment. There were 63 and 16 native and prosthetic valve endocarditis cases, respectively. They included 27 cases of culture negative endocarditis. With performing valve replacement or repair, 28 of the patient underwent a more aggressive surgical option, for example, aortic root replacement or reconstruction, or heart base reconstruction etc. Result: There were statistical relationships between the in-hospital mortality and staphylococcal infection, urgent-based operation and operation during the active phase of endocarditis. Wide debridement and aggressive reconstruction were not related to either the post operative mortality or the early morbidity. Culture negative endocarditis was not related to the postoperative mortality and morbidity. Conclusion: Physicians must pay attention to patients' medical treatment during the preoperative period of the infective endocarditis. If surgery is considered for treating infective endocarditis, it should be performed before the downhill course of the disease so that the surgical outcome is improved. Wide debridement and more aggressive reconstruction are also warranted.
Optimum doses The optimum dose that may be defined as the dose below the maximum permissible dose, yet would bring about a significant storage life extension at refrigerated temperatures, varied with species of fish as well as with the postirradiation storage temperatures. Thus the dose of 0. 1 Mrad was considered to be optimum for the croaker and yellow corvenia at $0^{\circ}C$, while at $5^{\circ}C$ the dose of 0.2 Mrad would be suitable for both species. The roundnose flounder was more radiosensitive and even at the dose of 0.1 Mrad a slight irradiation odor was detected immediately after the radiation treatment. Such degree of irradiation odor disappeared upon storage, therefore, the dose of 0.1 Mrad was considered to be optimum for the roundnose flounder at both $0^{\circ}\;and\;5^{\circ}C$. Storage life extension The croaker meats irradiated at 0.1 Mrad could be held at $0^{\circ}C$ as long as 5 weeks in good acceptable conditions, while the unirradiated control became unacceptable within 2 weeks-3-4 for extension of storage life at $0^{\circ}C$. At the storage temperature of $5^{\circ}C$, the storage life of 0.2 Mrad irradiated samples was extended from less than one week to 4 weeks--4-5 fold extension. The storage life extension of 0.1 Mrad irradiated yellow corvenia at $0^{\circ}C$ was from less than 2 weeks for the unirradiated to 4 weeks-approximately a-s folds and that of 0.2 Mrad irradiated samples stored at $5^{\circ}C$ was from 5 days to 3 weeks 4-5 folds. The roundnose flounder meats irradiated at 0.1 Mrad could held at $0^{\circ}C$ for 3-4 weeks as compared to less than 1 week for the unirradiated and at $5^{\circ}C$ the storage life could be extended from less than 3 days to up to 3 weeks. Thus the storage life extension by 4-5 folds and by 6-7 folds was possible at $0^{\circ}C\;and\;5^{\circ}C$ storage, respectively. Postirradiation storage microbiology and biochemistry In general 10 fold reduction of initial microflora was realized as a result of irradiating fish samples at 0.1 Mrad. The extent of microflora reduction increased with increasing doses applied, but not proportionately dependent. The microbial growth in the irradiated was severely retarded during the subsequent storage period, lagging far behind that of the irradiated control samples except in the late storage phase, when the levels of microflora of the irradiated either approached to or rose above the levels of the unirradiated. The microbiological changes caused by irradiation was reflected in the pronounced suppression of TVB and TMA accumulation during the storage period. This suggests that irradiation treatment brought about both quantitative and qualitative changes in microflora initially present and it is reasonable to suggest that the microflora removed by irradiation in fact represent most of the flora capable of producing TVB and TMA in normal fish spoilage process.
Lee, Dong-jun;Kim, Jung Kon;Jeong, Kwang-Hwa;Cho, Won-Mo;Ravindran, B.
Journal of the Korea Organic Resources Recycling Association
/
v.24
no.3
/
pp.63-74
/
2016
The objective of this study was to investigate the variations of physico-chemical properties during the swine manure composting, sawdust as the bulking agent was composted at different points (Top layer, Side of middle layer, Bottom layer). Air suction system with constant bottom aeration in bench scale reactors (30 L). The highest temperature was reached in the range of $58^{\circ}C$ to $62^{\circ}C$ on $3^{rd}$ day and this thermophilic phase (> $50^{\circ}C$) was continued for 3 days in all the treatment mixtures. However, the temperature was gradually decreased to room temperature at the end of 60 day composting process. Except control, the discharged ammonia ($NH_3$) was a maximum in the treatment order of Top layer>Bottom layer>Side of middle layer as 500 ppm, 162 ppm and 120 ppm, respectively, on the $4^{th}$ day and showing that Top layer point Air suction produce much more ammonia content than the other point. During the composting process, the total Kjeldahl nitrogen (TKN) was gradually increased due to the mass loss in the composting mixtures. At the same time, C/N ratio was decreased to Top layer, 13; Side of middle layer, 12 and Bottom layer, 13 at Air suction points. The significant reduction of C/N ratio in all different air suction system when manure was matured. The $NH_4-N$ to $NO_3-N$ ratio was recorded as 10.52 at the initial stage of the compost mixtures and reduced to 0.97 (Top layer), 0.70 (Side of middle layer), 3.2 (Bottom layer) because of manure decomposition. The overall results revealed that Top layer and Side of middle layer Air suction is a suitable option when compared other point for high quality composts.
The concentrations of quinolones (oxolinic acid; OXA, norlloxacin: NRF & ciprofloxacin: CPF) after oral administration of single doses (20 mg/kg B.W.) were investigated in carp (Cyprinus carpio) kept in freshwater at 20-23$^{\circ}C$. The distribution of the drug was studied after treatment. At points timed, from 1 h to 96 hrs after administration, blood (B), liver (L), kidney (K) and muscle (M) from 5 individuals in each group were collected for analyse with microbiological bioassay method. The peak concentrations were measured at 8 h (L), 12 h (B and K) and 24 h (M) after administration regardless of treated drugs. Considerably high concentrations of CPF (13.8-19.6${\mu}g/m{\ell}$) NRF (11.8-16.9${\mu}g/m{\ell}$) and OXA (10.8-13.9 ${\mu}g/m{\ell}$) were revealed during the 24 h. At the last time point of the experiment (96 h), concentrations of all three quinolones were: OXA, 2.3-6.3 ${\mu}g/m{\ell}$ ; NRF, 3.1-4.5 ${\mu}g/m{\ell}$ ; CPF, 3.0-5.5${\mu}g/m{\ell}$ in samples. The concentrations decreased subsequently, indicating a first rapid redistribution, followed by a slow phase of elimination. The steady state was observed in blood (12-36 h), liver (12-96 h) and muscle (36-96 h) after the initiation of treatment with OXA. Concerning the compartmental concentrations, (L, K. and M/B concentration ratio), the fluctuation of the ratio was founded at different time points, among drugs. For CPF, highest tissue ratios were prolonged in the order of L>K>M (0.65-1.2/0.82-0.93/1.0-1.7) during the experiments. On the other hand, NRF presented L>K>M (0.65-1.3/0.86-1.0) till 24 h, but L>M>K (0.89-1.26) at 36-96 h. OXA showed L>K>M (0.95-2.1) at 1-8 h, M>K>L (0.51-1.0) at 12-36 hand M>L>K (1.0-2.3) at 48-96 h, respectively.
Oh Sin Geun;Yang Kwang Mo;Hur Won Joo;Yoo Young Hyun;Suh Hong Suk;Lee Hyung Sik
Radiation Oncology Journal
/
v.20
no.4
/
pp.367-374
/
2002
Purpose : To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile acid derivative (HS-1200). Materials and Methods : Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-29 cells were examined using a frypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. Results : The dose required for the half maximal inhibition $(IC_{50})$ of the HT-29 cell growth was $100\~150\;{\mu}M$ of HS-1200. Several changes, associated with the apoptosis of the HT-29 cells, were reveal by the agarose gel eletrophoresis, TUNEL assays and Hoechst staining, following their treatment with $100\;{\mu}M$ of HS-1200. HS-1200 treatment also induced caspase-3, PARP and DFF degradations, and the western blotting showed the processed caspase-3 p20, PARP p85 and DFF p30 and p11 cleaved products. Mitochondrial events were also demonstrated. The cytochrome c staining indicated that cytochrome c had been released from the mitochondria in the HS-1200 treated cells. The mitochondrial membrane potential $(\Delta\Psi_m)$ was also prominently decreased in the HS-1200 treated cells. Conclusion : These findings suggest that the HS-1200 - induced apoptosis of human colon cancer cells (HT-29) is mediated via caspase and mitochondrial pathways.
Ryu, Young Hyo;Uhm, Han Sup;Park, Gyung Soon;Choi, Eun Ha
Journal of the Korean Vacuum Society
/
v.22
no.2
/
pp.55-65
/
2013
Sterilization of Neurospora crassa has been investigated in this research by using a surface air plasma with dielectric barrier discharged (DBD) structure under atmospheric pressure. The sinusoidal alternating current has been used in this experiment with discharge voltage of 1.4~2.3 kV. The phase difference between the voltage and current signals are found to be almost 80 degree due to the capacitive property of dielectric barrier. Temperature on the biomaterials has been minimized by radiating the heat with the air cooling system. It is noted that the substrate temperature remains under 37 degree for plasma exposure time of 10 minutes with operation of cooler system. It is found that the ozone, $O_3$, has been measured to be about 25~30 ppm within 1 cm region and to be about 5 ppm at the 150 cm downstream region away from the suface plasma. It is also noted that the nitric oxide, NO, and nitric dioxide, $NO_2$, are not nearly detected. Germination rate and mitochodrial activity of Neurospora crassa immersed in the deionized water have been found to be drastically decreased as the plasma treatment time and its electrical power are increased in this experiment. Here, the mitochondrial activity has been analyzed by MTT (3-(4,5-dimethy lthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. However, sterilization of Neurospora crassa immersed in the Vogel's minimal media has been found to be low by plasma treatment, which is caused by surrounding background solution. This research shows the sterilization possibility of Neurospora crassa by using the noncontated surface DBD plasma, which is different from the plasma jet. This is mainly attibuted to the reactive species generated by the surface plasma, since they play a major role for inhibition of micobes such as Neurospora crassa.
The removal yield of dissolved organic matter in drinking water by biological activated carbon (BAC) process was investigated. The tested processes wer raw water-AC process (BAC1), raw water-ozonation-BAC process (BAC2), and raw water-ozonation-coagulation/sedimentation-BAC process (BAC3). The amounts of organic matter was measured as dissolved organic carbon (DOC), ulta-violet radiation at 254 nm wavelength ($UV_{254}$), total nitrogen (T-N), ammonia nitrogen (NH_3$-N), and total phosphate (T-P). As a results, 30.7% DOC was removed by BAC2 process, which showed higher removal efficiency than BAC1 or BAC3 processes. The removal yield of $UV_{254}$ in BAC1, BAC2, and BAC3 processes were observed as 45.3%, 44.6%, 58.4%, respectively. And the removal yield of ammonia nitrogen were 66%, 81%, 29% in each BAC processes. The optimal empty bed contact time (EBCT) of BAC processes was estimated as 10 minute. This study has shown that BAC process combined with ozone treatment was efficient for removing dissolved organic matter in water.
Objective: The aim of this study was to compare GnRH antagonist and agonist flare-up treatment in the management of poor responder patients. Methods: One hundred forty-four patients from Jan. 1, 2002 to Aug. 31, 2005 undergoing IVF/ICSI treatment who responded poorly to the previous cycle (No. of oocyte retrieved$\leq$5) and had high early follicular phase follicle stimulating hormone (FSH>12 mIU/ml were selected. Seventy-five patients received agonist flare-up protocol and 71 patients received antagonist protocol. We analyzed the number of oocytes retrieved, number of good embryos (GI, GI-1), total dose of hMG administered, implantation rate, cycle cancellation rate, pregnancy rate, live birth rate. Results: The cancellation rate was high in antagonist protocol (53.5% vs. 30.1%). The number of oocyte retrieved, the number of good embyos were high in agonist flare-up group. There was no statistical difference between GnRH agonist flare up protocol and GnRH antagonist protocol in implantation rate (14.5%, 10.1%), clinical pregnancy rate per transfer (29.4%, 21.2%) and live birth rate per transfer (21.6%, 18.2%). Although the result was not statistically significant, GnRH agonist flare up group showed a nearly doubled pregnancy rate and live birth rate per initial cycle than GnRH antagonist group. Conclusions: The agonist flare-up protocol appears to be slightly more effective than the GnRH antagonist protocol in implantation rate, pregnancy rate, live birth rate but shows statistically no significance. Agonist flare-up protocol improved the ovarian response in poor responders. However, based of the result of the study, we can expect improved ovarian response in poor responders by GnRH agonist flare up protocol.
Lim, Eun Gyeong;Kim, Guen Tae;Kim, Bo Min;Kim, Eun Ji;Ha, Sung Ho;Kim, Sang-Yong;Kim, Young Min
Journal of Life Science
/
v.26
no.6
/
pp.663-672
/
2016
The Cnidium monnieri (L.) Cusson is an annual plant distributed in China and Korea. The fruit of C. monnieri is used as a medicinal herb that is effective for the treatment of carbuncle and pain in female genitalia. However, the anti-cancer effects of CME have not yet been reported. In this study, we assessed the apoptotic effects and cell cycle arrest effects of ethanol extracts from C. monnieri on HCT116 colon cancer cells. The results of an MTT assay and LDH assay demonstrated a decrease in cell viability and the cytotoxic effects of CME. In addition, the number of apoptotic body and the apoptotic rate were increased in a dose-dependent manner through Hoechst 33342 staining and Annexin V-PI double staining. In addition, cell cycle arrest occurred at the G1 phase by CME. Protein kinase B (Akt) plays an important role in cancer cell survival, growth, and division. Akt down-regulates apoptosis-mediated proteins, such as mammalian target of rapamycin (mTOR), p53, and Glycogen Synthase kinase-3β (GSK-3β). CME could regulate the expression levels of p-Akt, p-mTOR, p-GSK-3β, Bcl-2 family members, caspase-3, and PARP. Furthermore, treatment with CME, LY294002 (PI3K/Akt inhibitor), BIO (GSK-3β inhibitor), and Rapamycin (mTOR inhibitor) showed that apoptotic effects occurred through the regulation of the AKT/mTOR/GSK-3β signaling pathway. Our results demonstrated CME could induce apoptosis and cell cycle arrest in HCT116 colon cancer cells.
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