Phosphorus transfer from agricultural soils to surface waters is an important environmental issue. Fly ash and phospho-gypsum which are industrial by-product were investigated as a means of reducing dissolved phosphorus in arable soil. To determine the optimum mixing ratio of fly ash(FA) and phospho-gypsum(PG) for reducing dissolved reactive P(DRP) in soil, various mixture ratio of FA and PG were mixed with two soil. The DRP content and pH in soils were analysed after 3 weeks incubation under flooding condition. Although DRP content in soils was significantly decreased by FA-PG mixture compared with control, there were no significant difference among the FA and PG mixture ratio of 75:25, 50:50, and 25:75. The mixture of 75% FA and 25% PG was selected for field test. A field experiment was carried out to evaluate the reducing DRP content in paddy soil to which 0(NPK), 20(FG 20), 40(FG 40), and 60(FG 60) Mg $ha^{-1}$ of the mixture were applied. The DRP content was reduced by 31% at the application rate of 60 Mg $ha^{-1}$. In contrast to deceasing DRP, Ca-P content increased significantly with the mixture application rate. After rice harvesting, available $SiO_2$, P, and exchangeable Ca content in soil increased significantly with application rate due to high content of Si, P, and Ca in the mixture. Mixtures of fly ash and gypsum should reduce P loss from paddy soil and increase soil fertility.
The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.
Ok, Ji Un;Ha, Dong Uk;Lee, Shin Ja;Kim, Eun Tae;Lee, Sang Suk;Oh, Young Kyun;Kim, Kyoung Hoon;Lee, Sung Sill
Journal of Life Science
/
v.22
no.10
/
pp.1324-1329
/
2012
The objective of this study was to evaluate the in vitro effects of organic acids on methane emission and ruminal fermentation characteristics. We expected our methodology to result in a decrease of methanogens attached to the surface of rumen ciliate protozoa by addition of organic acids and in particular a decrease in methane emission. A fistulated Holstein cow of 650 kg body weight was used as a donor of rumen fluid. Organic acids (aspartic acid, fumaric acid, lactic acid, malic acid, and succinic acid) known to be propionate enhancers were added to an in vitro fermentation system and incubated with rumen fluid. The microbial population, including bacteria, protozoa, and fungi, were enumerated, and gas production, including methane and fermentation characteristics, were observed in vitro. Organic acids appeared to affect the rumen protozoan community. The rumen protozoal popuation decreased with the addition of aspartic acid, fumaric acid, lactic acid, and malic acid. In particular, the methane emission was reduced by addition of lactic acid. The concentration of propionate with all organic acids that were added appeared to be higher than that of the control at 12 h incubation. Addition of organic acids significantly affected rumen bacteria and microbial growth. The bacteria in added fumaric acid and malic acid was significantly higher (p<0.05) and protozoa was significantly lower (p<0.05) than that of the control. Microbial growth with the addition of organic acids was greater than the control after 48 h incubation.
This study was designed to get basic data about AIDS educational program development of students of university, especially for students of the department of Emergecy Medical Technology. The objective of this study was to identify the level of AIDS related Knowledge and Attitude of freshmen of university. The subject for this study consisted of 2022 male and female students who entered in 2005to Gongju National university in Chungnam province. Data was colledted by self-reporting questionaire consist of 66 items on 20th, February, 2005. The findings of this study can be summarized as follows. 1) The mean overall knowledge score was estimated to be 70.22%. 2) The knowledge score about definition of AIDS was high(93.2%). but knowledge score about progression and incubation period was low(48%). 3) AIDS related knowledge about diagnosis with blood was was high(91.2%), but those about period of antibody formation was relatively low(66.2%). 4) The score about latent appearance of AIDS-related symptoms was hlgh(93.7%), but those of apprehension of individual symptom was very low(57.5%). 5) Percentage of correctly answered respondent about transmission with needle and transfusion was very high(>94%), but the score of transmission through the anal and oral sex was relatively low(75-79%). 6) The knowledge score about prevention with condom was high(89.5%), but misconception of disinfection and vaccination was also high. 7) Acknowledgement about utility of consultation, information, treatment was very low (10-17%). In the end, the study concludes that it is necessary to develop comprehensive AIDS education programs to improve knowledge about the disease as well as to allay the fears and anxiety of the contact.
Kim, Dong-Myung;Choi, Cha-Yong;Ahn, Jin-Ho;Kim, Tae-Wan;Kim, Nam-Young;Oh, In-Suk;Park, Chang-Gil
Biotechnology and Bioprocess Engineering:BBE
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v.11
no.3
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pp.235-239
/
2006
Due to recent advances in genome sequencing, there has been a dramatic increase in the quantity of genetic information, which has lead to an even greater demand for a faster, more parallel expression system. Therefore, interest in cell-free protein synthesis, as an alternative method for high-throughput gene expression, has been revived. In contrast to in vivo gene expression methods, cell-free protein synthesis provides a completely open system for direct access to the reaction conditions. We have developed an efficient cell-free protein synthesis system by optimizing the energy source and S30 extract. Under the optimized conditions, approximately $650{\mu}g/mL$ of protein was produced after 2h of incubation, with the developed system further modified for the efficient expression of PCR-amplified DNA. When the concentrations of DNA, magnesium, and amino acids were optimized for the production of PCR-based cell-free protein synthesis, the protein yield was comparable to that from the plasmid template.
Kim, Sun Ha;Shim, Kyu-Chan;Lee, Hyun-Sook;Le, Anh Quynh;Ahn, Sang-Nag
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.116-116
/
2017
Low-temperature is one of the environmental stress factors that affect plant growth and development and consequently limit crop productivity. The control of seed germination under low-temperature is organized by many genes which are called quantitative trait loci (QTLs). High germination rate for low-temperature is an important factor of growing rice. Previously, we identified a major QTL controlling low-temperature germinability in rice using 96 introgression lines (ILs) derived from a cross between Oryza rufipogon (Rufi) and the Korean japonica cultivar, 'Hwaseongbyeo (HS)'. A $BC_3F_7$ line (TR5) showed better low-temperature germinability than its recurrent parent. TR5 was crossed with HS to develop a segregating F2:3 populations for the target QTL. Six SSR markers polymorphic between HS and Rufi were used to screen and fine map the qLTG1. The qLTG1 on chromosome 1, which accounted for 55.5% of the total phenotypic variation, confirmed that Rufi allele enhanced the low-temperature germinability. Intervals between markers CRM16 and CRM15, four candidate genes were identified. The identified candidate genes, which are encoded by a protein of unknown function, showed their direct involvement on seed germination at low-temperature. To identify genes targeted by qLTG1, we investigated the expression profiles of these candidate genes and germination behavior of qLTG1 under different stress conditions and compared to HS, Rufi, and TR5 at $13{\pm}2^{\circ}C$ for 3 days after incubation. Furthermore, transgenic rice plants will also be developed to conduct a detailed investigation on low-temperature germinability. Hence, the QTL for low-temperature germinability would be useful in rice breeding programs especially in the development of lines possessing low-temperature germinability.
Despite increasing success rate of IVF, poor response to ovarian stimulation remains a problem. So, attempts to improve ovarian responses, for example, by using combined gonadotropin-releasing hormone analogue(GnRH-a) and human menopausal gonadotropin(hMG) have shown limited success. It is reported that response of granulosa cells in vitro to FSH is stimulated by co-incubation with IGF-l, and IGF-l production can be increased by growth hormone. This suggest that combination regimen of G.H. and hMG may augment follicle recruitment. In fifteen patients who had previous history of poor ovarian response to gonadotropin stimulation after pituitary suppression with mid -luteal GnRH-a, the effectiveness of cotreatment with G.H. in IVF program was evaluated using a combination regimen of G.R. and hMG at Korea University Hospital IVF Clinic. Ovarian responses to gonadotropin stimulation in control and GH-treated cycles assessed by total dose and duration of hMG treatment, follicular development and peak $E_2$ level, number of eggs retrieved, and fertilization rates were also assessed. In each group, serum and follicular fluid IGF-1 concentrations on day of egg collection were measured by RIA after acidification and extraction by reveresed phase chromatography. Patients receiving G.H. required fewer days and ampules of gonadotropins, developed more oocytes, and more embryos transferred. But, the differences were not statistically significant, except the duration of hMG treatment. Our data showed a significantly higher concentration of IGF-l in the serum, not in the follicular fluid, of patients treated with G.H. compared with control group. These data suggest that growth hormone treatment does not improve the ovarian response in women with limited ovarian reserve to gonadotropin stimulation for IVF.
The purpose of this study is to characterize canine mesenchymal stem cells (MSCs) derived from bone marrow (BM) for use in research on the applications of stem cells in canine models of development, physiology, and disease. BM was harvested antemortem by aspiration from the greater tubercle of the humerus of 30 normal beagle dogs. Canine BM-derived MSCs were isolated according to methods developed for other species and were characterized based on their morphology, growth traits, cell-surface antigen profiles, differentiation repertoire, immunocytochemistry results, and neurotrophic factor expression in vitro. The canine MSCs exhibited a fibroblast-like morphology with a polygonal or spindle-shaped appearance and long processes; further, their cell-surface antigen profiles were similar to those of their counterparts in other species such as rodents and humans. The canine MSCs could differentiate into osteocytes and neurons on incubation with appropriate induction media. RT-PCR analysis revealed that these cells expressed NGF, bFGF, SDF-1, and VEGF. This study demonstrated that isolating canine MSCs from BM, stem-cell technology can be applied to a large variety of organ dysfunctions caused by degenerative diseases and injuries in dogs. Furthermore, our results indicated that canine MSCs constitutively secrete endogenous factors that enhance neurogenesis and angiogenesis. Therefore, these cells are potentially useful for treating dogs affected with various neurodegenerative diseases and spinal-cord injuries.
This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.
Conjugated linoleic acid (CLA) exhibits several beneficial biological activities including anticarcinogenesis and body-fat reduction. Now, we report that CLA ameliorated the oxidative stress in rat cardiomyoblast cells, H9c2, treated with hydrogen peroxide ($H_2O_2$). Cells were cultured in DMEM/F-12 media at $37^{\circ}C$ with humidified atmosphere of 5% $CO_2$. The cells, cultured for 48 hrs, were seeded at a density $3.5{\times}10^3$ cell/well in a 24 well-plate and incubated for 24 hr. Using these cells, two experiments were performed: the cytotoxicity test of CLA (10, 20, 30, 40, and $50{\mu}Ms$), and the oxidative stress amelioration test of CLA (20 and $50{\mu}Ms$) against cells treated with $H_2O_2$ (10 and 50 ${\mu}Ms$) for 1 and 2 hrs. CLA enhanced the growth of H9c2 cells at any concentrations of CLA and at any incubation times (up to 6 days), indicating that CLA acts as a growth stimulant. No protective effect of CLA (20 and $50{\mu}Ms$) was seen in cells treated $50{\mu}M$$H_2O_2$ for 1 and 2 hr, but these CLA concentrations ameliorated (p<0.05) the adverse effect of $10{\mu}M$$H_2O_2$ in cells treated for 1 hr. These CLA concentrations significantly (p<0.05) reduced the proportion of apoptotic cells, relative to control cells. These results suggest that CLA protected H9c2 cells from the oxidative stress of $H_2O_2$ through the suppression of cell apoptosis and could be a useful compound for the prevention of cardiac diseases caused by oxidative stress.
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