Edaravone (Eda) is a potent scavenger of inhibiting free radicals including hydroxyl radicals ($H_2O_2$). Reactive oxygen species (ROS) such as $H_2O_2$ can alter most kinds of cellular molecules such as lipids, proteins and nucleic acids, cellular apoptosis. In addition, oxidative stress from over-production of ROS is involved in the defective embryo development of porcine. Previous study reported that Eda has protective effects against oxidative stress-like cellular damage. However, the effect of Eda on the preimplantation porcine embryos development under oxidative stress is unclear. Therefore, in this study, the effects of Eda on blastocyst development, expression levels of ROS, and apoptotic index were first investigated in preimplantation porcine embryos. After in vitro fertilization, porcine embryos were cultured for 6 days in PZM medium with Eda ($10{\mu}M$), $H_2O_2$ ($200{\mu}M$), and Eda+$H_2O_2$ treated group, respectively. Rate of blastocyst development was significantly increased (P<0.05) in the Eda treated group compared with only $H_2O_2$ treated group. And, we measured intracellular levels of ROS by DCF-DA staining methods and investigated numbers of apoptotic nuclei by TUNEL assay analysis is in porcine blastocyst, respectively. Both intracellular ROS levels and the numbers of apoptotic nucleic were significantly decreased (P<0.05) in porcine blastocysts cultured with Eda ($10{\mu}M$). More over, the total cell number of blastocysts were significantly increased (P<0.05) in the Eda-treated group compared with untreated group and the only $H_2O_2$ treated group. Based on the results, Eda was related to regulate as antioxidant-like function according to the reducing ROS levels during preimplantation periods. Also, Eda is beneficial for developmental competence and preimplantation quality of porcine embryos. Therefore, we concluded that Eda has protective effect to ROS derived apoptotic stress in preimplantation porcine embryos.
The objective of this study was to evaluate the efficiency of the conventional slow freezing and vitrification methods for cryopreserving in vivo and in vitro-produced bovine embryos. Morphology of post-thawed embryos was evaluated and normal embryos were used for successive culture for 72 h. In experiment I, In embryo viability, There was no significant differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos(89.6% vs. 81.5%). whereas hatched-BL and total cell number rates was significantly higher (p<0.05) for in vivo-derived embryos (76.9%, 136${\pm}$3.6 vs. 43.4%, 107${\pm}$3.8). In experiment II, There was no significant differences in blastocyst re-expansion and Expansion-BL rates were found between in slow freezing and vitrification methods (91.3% vs. 85.7% and 71.4% vs. 75.0%, respectively). in conclusion, These results suggested that the field application for bovine embryo transfer is in part supported by improvements of technologies in embryo conventional slow freezing and vitrification cryopreservation.
Journal of the Korean Society of Food Science and Nutrition
/
v.33
no.10
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pp.1709-1714
/
2004
The changes of microflora and enzyme activities in Alaska pollack sikhae were evaluated in 3 different temperature conditions, 5$^{\circ}C$, 2$0^{\circ}C$ and alternating temperature (stored at 5$^{\circ}C$ after 10 days of fermentation at 2$0^{\circ}C$), respectively. The number of proteolytic bacteria and 2 lactic acid bacteria including Lactobacillus sp. and Pediococcus sp. increased rapidly up to 10 days and composed major portion of total viable cell (TVC) in sikhae fermented at 2$0^{\circ}C$, whereas those of TVC were occupied by Lactobacillus sp., Pediococcus sp. and yeast after 10 days of fermentation. The major species of microflora in sikhae fermented at alternating temperature were, composed of Lactobacillus, Pediococcus and Streptococcus after 10 days of fermentation. Especially, Leuconostoc sp. was kept up to 27 days at 5$^{\circ}C$ than other temperature conditions (16 days). The activities of protease and lipase in acidic region (pH 3.0) were higher at 2$0^{\circ}C$ than at 5$^{\circ}C$ due to sensitivity of temperature, although those of protease and lipase in neutral region (pH 7.0) were not found any differences in both temperatures. Changing temperature condition from 2$0^{\circ}C$ to 5$^{\circ}C$ in alternating temperature inactivated protease activity, whereas lipase activity was still maintained during fermentation.
A study was made of the concentration of the naturally occurring radionuclide $^{210}Po$ in seawater, plankton and fishes collected from Yongil Bay, Korea, in summer 1993. The mean value of the $^{210}Po$ concentration in sea water was determined to be about 1.9 mBq/l, with the proportion of the dissolved forms being about $46\%$. The mean $^{210}Po$ concentration factor in the centric diatom Skeletonema costatum, comprises > $95\%$ of the total phytoplankton cell number, was $7.6\times10^4$. Of five species of zooplankton examined, four species (two copepods and two mysids) had $^{210}Po$ concentrations in the range of $^{210}Po$ mBq/g dry weight. A copepod Labidocera bipinnata, however, was exceptional with a particularly high $^{210}Po$ level of 2,070 mBq/g. This indicates that L. bipinnata is primarily a raptorial feeder. The concentration of $^{210}Po$ in the pyloric caecum of pelagic fishes was 2,979-3,811 mBq/g, with the concentration factors of $3.4-4.3\times10^6$. The food chain concentration of $^{210}Po$ occurred as follows: phytoplankton < filter feeding copepods < omnivorous mysids < raptorial copepod < pelagic fishes.
Lee Sang Hun;Lee Seung Eon;Lee Si Hyeong;Shin Jo Young
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.2
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pp.528-533
/
2004
The immune system acts to protect the host from infectious agents that exist in the environment and from other noxious insults. The immune system has two functional divisions: the innate and the acquired. Both components involve various factors such as cytokines. A number of methodologies exist to assess aspects of immune function. There are large inter-individual variations in many immune functions even among the healthy. Genetics, age, gender, smoking habits, habitual levels of exercise, alcohol consumption, diet, stage in the female menstrual cycle, stress, history of infections and vaccinations, and early life experiences are likely to be important contributors to the observed variation. While it is clear that individuals with immune responses significantly below 'normal' are more susceptible to infectious agents and exhibit increased infectious morbidity and mortality, it is not clear how the variation in immune function among healthy individuals relates to variation in susceptibility to infection. Oriental medicine is an important factor contributing to immune competence. The author investigated the immune enhancement effects of Bojungikkitanggami-bang (BITB). The forced swimming test (FST) has been used as a screening model for new immune enhancement agents. In the present study, the author investigated the effects of BITB on FST and blood biochemical parameters related to fatigue, glucose (Glc); blood urea nitrogen (BUN); lactate dehydrogenase (LDH); creatinine; and total protein (TP). The author found that BITB (1 g/kg) significantly reduced the immobility time in the FST compared to the control. In addition, the contents of Glc, LDH, BUN, TP in the blood serum were increased in BITB (1g/kg)-fed group. Also, the author investigated the effects of BITB on the production of cytokines in human T-cell line, MOLT-4 cells. BITB (1 mg/ml) significantly increased the interferon (IFN)-vproduction compared with media control (about 2.2-fold for IFN-γ) at 24 h. However, BITB has not affect the production of IL-2 and IL-4. In addition, BITB increased the protein expression level of IFN-γ in MOLT-4 cells. Thus, BITB may have therapeutic value in generating or enhancing immune function in a clinical setting.
To determine the dominant bacterial species during the Saeu-jeotgal ripening process, the cultivable bacterial population was examined over a 135-day period using six different growth media. The greatest numbers of bacteria were identified when marine agar was used for culture, with maximum cell density identified at day 65 (2.51 × 107 colony forming units/g). Over the course of 135 days, the bacterial diversity was analyzed eight times. A total of 467 isolates, comprising 87 species from 42 genera, as well as 16 isolates belonging to previously unknown species, were identified. The number of species detected decreased from 39 at day 1 to 13 at day 135. The order of dominance at the genus level was as follows: Staphylococcus, Salimicrobium, Kocuria, and Psychrobacter. Staphylococcus and Salimicrobium accounted for 2% of the diversity at day 1, and then increased to 39% and 36%, respectively, at day 135. The dominant species Staphylococcus equorum, Salimicrobium salexigens, and Kocuria palustris accounted for 23.6%, 16.1%, and 10.9% of all isolates, respectively. Importantly, both St. equorum and Sm. salexigens remained viable at a NaCl concentration of 21% (w/v), which indicates their strong involvement in the ripening of Saeu-jeotgal.
Yogurt was prepared with different substitution ratio [10, 30, and 50% (w/w)] of skim milk with enzymatically hydrolyzed purple sweet potato powder (EHPSPP) and fermented at $40^{\circ}C$ for 15 h. Fermentation characteristics and antioxidant activities of the yogurt were evaluated in terms of acid production (pH and titratable acidity) and lactic acid bacterial counts and DPPH radical scavenging activity, respectively. After 15 h fermentation, titratable acidity of EHPSPP yogurt was 0.80-0.89% and was lower than that (0.93%) of yogurt made without EHPSPP. The acid production and the number of viable lactic acid bacterial cell decreased with increasing the substitution ratio. The sensory score of EHPSPP yogurt prepared with 30% substitution ratio showed the highest values in taste and overall acceptability among the tested yogurt preparations. DPPH radical scavenging activity increased with increasing the substitution ratio in yogurt fermented for 12 h. The total phenolic content of 30% EHPSPP yogurt was 40% higher than that of skim milk yogurt. These results suggest that EHPSPP can be used as substituent of skim milk and the optimum substitution ratio is around 30%.
Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG ($33.1{\pm}9.6$ vs $21.7{\pm}8.3%$). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP-treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The mRNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.
The $Na^+-and\;K^+-induced\;Ca^{++}$ release was measured isotopically by millipore filter technique in pig heart mitochondria. With EGTA-quenching technique, the characteristics of mitochondrial $Ca^{++}-pool$ and the sources of $Ca^{++}$ released from mitochondria by $Na^+\;or\;K^+$ were analyzed. The mitochondrial $Ca^{++}-pool$ could be distinctly divided into two components: internal and external ones which were represented either by uptake through inner membrane, or by energy independent passive binding to external surface of mitochondria, respectively. In energized mitochondria, a large portion of $Ca^{++}$was transported into internal pool with little external binding, while in de-enerigzed state, a large portion of transported $Ca^{++}$ existed in the external pool with limited amount of $Ca^{++}$ in the internal pool which was possibly transported through the $Ca^{++}-carrier$ present in the inner membrane. $Na^+$ induced the $Ca^{++}$ release from both internal pool and external pool and external binding pool of mitochondria. In contrast, $K^+$ did not affect $Ca^{++}$ of the internal pool, but, displaced $Ca^{++}$ bound to external surface of the mitochondria. When the $Ca^{++}-reuptake$ was blocked by EGTA, the $Ca^{++}$ release from the internal pool by $Na^+$ was rapid; the rate of $Ca^{++}-efflux$ appeared to be a function of $[Na^+]^2$ and about 8mM $Na^+$ was required to elicit half-maximal velocity of $Ca^{++}-efflux$. So it was revealed that $Ca^{++}-efflux$ velocity was particulary sensitive to small changes of the $Na^+$ concentration in physiological range. Energy independent $Ca^{++}-binding$ sites of mitochondrial external surface showed unique characteristics. The total number of external $Ca^{++}-binding$ sites of pig heart mitochondria was 29 nmoles per mg protein and the dissociation constant(Kd) was $34{\mu}M$. The $Ca^{++}-binding$ to the external sites seemed to be competitively inhibited by $Na^+\;and\;K^+$; the inhibition constant(Ki) were 9.7 mM and 7.1 mM respectively. Considering the intracellular ion concentrations and large proportion of $Ca^{++}$ uptake in energized mitochondria, the external $Ca^{++}-binding$ pool of the mitochondria did not seem to play a significant role on the regulation of intracellular free $Ca^{++}$ concentration. From this experiment, it was suggested that a small change of intracellular free $Na^+$ concentration might play a role on regulation of free $Ca^{++}$ concentration in cardiac cell by influencing $Ca^{++}-efflux$ from the internal pool of mitochondria.
Kochujang-gulbi, a Korean traditional food, was prepared with kochujang and freeze-dried gulbi slices. Kochujang-gulbi was treated with high hydrostatic pressure (200, 400 and 600 MPa) and gamma-irradiation (7, 10, 20 and 30 KGy) to improve its quality and shelf-life. The pH of high hydrostatic pressure and gamma-irradiation treated kochujang-gulbi samples did not significantly different compared to that of control. However, Hunter L value slightly increased, and Hunter a and b values decreased by high hydrostatic pressure and gamma-irradiation treatment. During storage, the pH and color of high hydrostatic pressure and gamma-irradiation treated kochujang-gulbi samples did not significantly changed. The VBN and TBA level decreased by the increase treatment of high hydrostatic pressure and then slowely increased during storage. The VBN and TBA level of kochujang-gulbi samples treated by the strong gamma ray emission slowely increased during storage. In particular, gamma-irradiation treatment was very effective to sterilize microorganisms when compared to that of high hydrostatic pressure in kochujang-gulbi products. In addition, the high hydrostatic pressure and gamma-irradiation treated kochujang-gulbi samples shown significantly lower total viable cell number than control for the 20 days of storage at room temperature with retort pouch packaging.
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