Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).
Journal of the Korean Institute of Oriental Medical Informatics
/
v.11
no.1
/
pp.52-57
/
2005
Objectives : Recurrent pregnancy loss occurs in approximately 0.5-3% of women. There are many studies concerning immunological factor recently. Therefore, aim of this study is to examine the latest trend of researches concerning recurrent pregnancy loss, and controlled experiment on animals about antiphospholipid antibody. Method : We referred Pubmed site by using searching word of 'recurrent pregnancy loss' (Limits : 2000.1-2004.3, animal) Results and conclusions : 1. We searched 29 papers. Immunological factor : 18 ( about antiphospholipid antibody : 10 ), Chromosomal abnormality : 6, The others : 5 2. In five papers about controlled experiment on animals, (1) Materials : 8-12 weeks old mice / 11.5-day old-rat embryos / New Zealand rabbit (2) Inductions : inject intraperitoneally with human IgG containing antiphospholipid antibodies / culture in a solution of 1 ml medium which contained IgG purified from sera of women / inject intradermally with cardiolipin (3) Treatments : inject intraperitoneally with complement component before / culture in a solution of 1 ml medium which contained IgG purified from sera of women with SLE and RPL or from healthy women / inject intradermally with TFX, 0.9% NaCl (4) Measurements : weight fetuses and placentas, calculate frequency of fetal resorption / after culture, examine the embryos / examine platelet counts, APTT and numbers of live and dead newborns, resorbed fetuses, body mass, newborn viability and survival rates.
Ding, Ning;Lu, Yanzhu;Cui, Hanmin;Ma, Qinyu;Qiu, Dongxia;Wei, Xueting;Dou, Ce;Cao, Ning
BMB Reports
/
v.53
no.3
/
pp.154-159
/
2020
We investigated the effects of physalin A, B, D, and F on osteoclastogenesis induced by receptor activator of nuclear factor κB ligand (RANKL). The biological functions of different physalins were first predicted using an in silico bioinformatic tool (BATMAN-TCM). Afterwards, we tested cell viability and cell apoptosis rate to analyze the cytotoxicity of different physalins. We analyzed the inhibitory effects of physalins on RANKL-induced osteoclastogenesis from mouse bone-marrow macrophages (BMMs) using a tartrate-resistant acid phosphatase (TRAP) stain. We found that physalin D has the best selectivity index (SI) among all analyzed physalins. We then confirmed the inhibitory effects of physalin D on osteoclast maturation and function by immunostaining of F-actin and a pit-formation assay. On the molecular level, physalin D attenuated RANKL-evoked intracellular calcium ([Ca(2+)](i)) oscillation by inhibiting phosphorylation of phospholipase Cγ2 (PLCγ2) and thus blocked the downstream activation of Ca2+/calmodulin-dependent protein kinases (CaMK)IV and cAMP-responsive element-binding protein (CREB). An animal study showed that physalin D treatment rescues bone microarchitecture, prevents bone loss, and restores bone strength in a model of rapid bone loss induced by soluble RANKL. Taken together, these results suggest that physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via suppressing the PLCγ2-CaMK-CREB pathway.
Ssanghwa-tang (SHT) is a traditional Korean herbal medicine widely prescribed to decrease fatigue following an illness. The purpose of this study was to investigate the effects of SHT on osteoclast differentiation in vitro, and on bone loss in ovariectomized (OVX) rats in vivo. SHT significantly reduced the receptor activator for the nuclear factor ${\kappa}B$ (NF-${\kappa}B$) ligand (RANKL)-induced tartrate-resistant acid phosphatase (TRAP) activity, and multinucleated osteoclast formation in RAW264.7 cells without affecting cell viability. In addition, SHT significantly attenuated RANKL-induced mRNA expression levels of c-Src and cathepsin K. To examine the in vivo effect of SHT on OVX-induced bone loss in OVX rats, we administered SHT (0.6 g/kg BID) orally to OVX rats for 12 weeks. SHT administration significantly blocked OVX-induced decrease of femoral bone mineral density (BMD) and femoral trabeculae in OVX rats. In conclusion, these results suggest that SHT treatment effectively prevents OVX-induced bone loss, and this effect may result from its inhibitory effect on osteoclast differentiation.
Journal of the Korean Society for Nondestructive Testing
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v.28
no.2
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pp.191-198
/
2008
Flow accelerated corrosion (FAC) phenomenon of low alloy carbon steels in nuclear power plant has been known as one of major degradation mechanisms. It has a potential to cause nuclear pipe rupture accident which may directly impact on the plant reliability and safety. Recently, the equipotential switching direct current potential drop (ES-DCPD) method has been developed, by the present authors, as a method to monitor wall loss in a piping. This method can rapidly monitor the thinning of piping, utilizing either the wide range monitoring (WiRM) or the narrow range monitoring (NaRM) technique. WiRM is a method to monitor wide range of straight piping, whereas NaRM focuses significantly on a narrow range such as an elbow. WiRM and NaRM can improve the reliability of the current FAC screening method that is based on computer modeling on fluid flow conditions. In this paper, the measurements by ES-DCPD are performed with signal sensitivity analyses in the laboratory environment for extended period and showed the viability of ES-DCPD for real plant applications.
Kim, Jinhee;Lee, Hyejin;Kang, Ki Sung;Chun, Kwang-Hoon;Hwang, Gwi Seo
Journal of Ginseng Research
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v.39
no.1
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pp.46-53
/
2015
Background: Glucocorticoids (GCs) are commonly used in many chemotherapeutic protocols and play an important role in the normal regulation of bone remodeling. However, the prolonged use of GCs results in osteoporosis, which is partially due to apoptosis of osteoblasts and osteocytes. In this study, effects of Korean Red Ginseng (KRG) on GC-treated murine osteoblastic MC3T3-E1 cells and a GC-induced osteoporosis mouse model were investigated. Methods: MC3T3-E1 cells were exposed to dexamethasone (Dex) with or without KRG and cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Realtime polymerase chain reaction was performed to evaluate the apoptotic gene expression; osteogenic gene expression and alkaline phosphatase (ALP) activity were also measured. Western blotting was performed to evaluate the mitogen-activated protein kinase (MAPK) proteins. A GC-induced osteoporosis animal model was used for in vivo study. Results and conclusion: The MTT assay revealed that Korean Red Ginseng (KRG) prevents loss of cell viability caused by Dex-induced apoptosis in MC3T3E1 cells. Real-time polymerase chain reaction data showed that groups treated with both Dex and KRG exhibited lower mRNA levels of caspase-3 and -9, whereas the mRNA levels of Bcl2, IAPs, and XIAP increased. Moreover, groups treated with both Dex and KRG demonstrated increased mRNA levels of ALP, RUNX2, and bone morphogenic proteins as well as increased ALP activity in MC3T3-E1 cells, compared to cells treated with Dex only. In addition, KRG increased protein kinase B (AKT) phosphorylation and decreased c-Jun N-terminal kinase (JNK) phosphorylation. Moreover, microcomputed tomography analysis of the femurs showed that GC implantation caused trabecular bone loss. However, a significant reduction of bone loss was observed in the KRG-treated group. These results suggest that the molecular mechanism of KRG in the GC-induced apoptosis may lead to the development of therapeutic strategies to prevent and/or delay osteoporosis.
We investigated the neuroprotective effect of an ethanol extract from Asparagus cochinchinensis (AC) against glutamate-induced toxicity in the HT22 hippocampal cell, which is an ideal in vitro model for oxidative stress. The neuroprotective effects of AC in HT22 cells were evaluated by analyzing cell viability, lactate dehydrogenase (LDH), flow cytometry for cell death types, reactive oxygen species (ROS), mitochondria membrane potential (MMP), and Western blot assays. In the cell death analysis, AC treatment resulted in significantly attenuated glutamate-induced loss of cell viability with a decrease in LDH release. AC treatment also reduced glutamate-induced apoptotic cell death. In the ROS and MMP analysis, AC treatment inhibited the elevation of intracellular ROS induced by glutamate exposure and the disruption of MMP. In oxidative stress-related proteins analysis, AC treatment inhibited the expression of poly ADP ribose polymerase and heme oxygenase-1 by glutamate. These results indicate that AC exerts a significant neuroprotective effect against glutamate-induced hippocampal damage by decreasing ROS production and stabilizing MMP. Thus, AC potentially provides a new strategy for the treatment of oxidative stress-related diseases.
The Journal of the Korean Society for Microbiology
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v.21
no.4
/
pp.435-439
/
1986
The viability of tubercle bacilli in the sputum specimens has been investigated after different periods of storage at diffeent temperatures and in the presence of different preservatives. No loss of culture positives was observed for one week storage at $4^{\circ}C$, but 9.8% 19.5%C, and 26.8% of sputums failed to yield positives on 2, 3, and 4 weeks of storage respectively. At $25^{\circ}C$ even one week storage made 19.5% of sputums fail to yield positive culture and 2, 3, and 4 weeks of storage made 36.6% 70.7%, and 90.2% of sputums fail to yield positive culture respectively. And contamination was unacceptably high beyond one week of storage at $25^{\circ}C$. Contamination of sputum specimens could be protected fairly well by 0.5% boric acid, by 5% trisodium phosphate or by 0.5% cetylpyridium chloride, but, except CP, the former two had no advantage at all to protect viability of tubercle bacilli over the specimens without preservative. The CP was much less harmful to the viability of tubercle bacilli than BA, yielding 61.0% and 31.7% of culture positives on 3 and 4 weeks of storage in the presence of CP, while BA yielded 34.1% and 4.9% of positives on the same respective periods of storage. Therefore CP may be useful to preserve sputums if it takes more than 2 weeks to transport them at the temperature of over $25^{\circ}C$.
The Journal of the Institute of Internet, Broadcasting and Communication
/
v.24
no.4
/
pp.155-165
/
2024
Software as a Service (SaaS) has become one of the fastest-growing software business models in recent years. Even during the economic downturn following the pandemic, the SaaS business has emerged as a crucial model for IT companies. The revenue structure of SaaS, which is based on the subscription economy model, ensures that users pay only for the services used. In other words, SaaS operates on a subscription-based billing model, thus providing subscribers access to software uploaded to cloud computers via the Internet. This study aimed to explore the manner by which software-solution firms have to counteract the decline in profit and loss sales caused by changing their business-model orientation from on-premise deployment software to subscription-based software. Additionally it analyzes a method for selecting a subscription-based pricing model and rapidly recovering the investment costs via quantitative business-viability assessment. By calculating subscription fees via a more quantitative business-viability evaluation instead of focusing on conventional business-planning methods that rely on qualitative methods, companies are expected to be equipped in providing services to customers at reasonable costs. This strategy will facilitate them in leading emerging growth sectors.
Experiments were conducted to investigate seed persistence of Chinese milk vetch(CMV) in naturally reseeded rice field in 2007~2009. The seed and pods with seeds were buried in rice field at 0, 5 and 10 cm depths and retrieved from the field at one to three month intervals from July to the following year March and determined change of seed dormancy and viability. In the second experiment, persistence of the CMV seeds in the naturally reseeded rice field at different tillage methods and soil depths were also investigated after rice harvest in autumn. Burial depths and durations affected recovery rate, dormancy and viability of CMV seed. The viability loss was faster and greater in the seed than the pod with seeds and on the soil surface than the 5 or 10 cm burial depths. The recovery rate of CMV seed was decreased starting from one month as seed burial and it was significantly decreased to 52~65% for the seed in September. However, unlike the seed burial, the nearly 100% CMV seeds were recovered for burial as pod with seeds even after four months burial in both 0 and 5 cm depths. However, the recovery rate was sharply declined to below 30% at October in 2007 in both seed and pods with seeds and in the 2008/2009 experiment. the 15~47% of CMV seeds still remained even after October. The CMV had high seed dormancy of 95%, showing only 4~5% germination at the beginning in June but the seed germination increased to 25 to 35% in seed and 55 to 61% in pod with seeds in September due to breakage of hard seed dormancy. The viability loss was faster in the seed than in the pod with seeds regardless of depths of placement in the soil base on decayed seeds. Also the seed placed on the soil surface lost viability faster than the 5~10 burial depths. On the other hand, field observation in the naturally reseeded CMV rice field showed that as many as 917~2,185 CMV seeds $m^2$ were from the 0~15 cm soil depth in the rotary tillage and 250~10,105 CMV seeds in minimum tillage treatmints. The recovered seed germinated 25~33%, 23~43% but still had high percentage of hard seed having 64~72% and 51~77% even after rice harvest in autumn. These results indicate that freshly harvested CMV seeds had high level of primary dormancy and the dormancy was gradually broken in soil with time during rice cultivation periods and appreciable number of CMV seeds remained even 4 month after burial in soil. CMV plant regenerated naturally from the remained seed bank at rice harvest time in autumn. The CMV seedling still emerged even after 2 years of continuous destructive killing of emerged CMV plant by rotary tillage in naturally reseeded CMV plant in rice field, indicating that CMV seeds do persistent as least two years in soil.
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