• Journal of Animal Reproduction and Biotechnology
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• v.38 no.2
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• pp.47-53
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• 2023

Despite numerous advances in in-vitro embryo production (IVP), many documented factors have been shown to influence the development of mammalian preimplantation embryos and the success of IVP. In this sense, elevated levels of reactive oxygen species (ROS) correlate with poor outcomes in assisted reproductive technologies (ART) due to oxidative stress (OS), which results from an imbalance between ROS production and neutralization. Indeed, excessive production of ROS compromises the structural and functional integrity of gametes and embryos both in vivo and in vitro. In particular, OS damages proteins, lipids, and DNA and accelerates cell apoptosis. Several in-vivo and in-vitro studies report an improvement in qualityrelevant parameters after the use of various antioxidants. In this review, we focus on OS and the source of free radicals and their effects on oocytes, sperm, and the embryo during IVP. In addition, antioxidants and their important role in IVP, supplementation during oocyte in vitro maturation (IVM), in vitro culture (IVC), and semen extenders were discussed. Nevertheless, various methods for determining the level of ROS in germ cells have been briefly described. Still, it is crucial to develop standardized antioxidant supplement systems to improve overall IVP success. Further studies should explore the safety, efficacy, mechanism of action, and combination of different antioxidants to improve IVP outcomes.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.3
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• pp.334-339
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• 2007

Reactive oxygen species (ROS) generate during electrical activation of oocytes which has detrimental effects on embryo survival when overwhelmed. The present study was designed to investigate the ability of L-ascorbic acid, a novel water soluble antioxidant, to reduce the ROS level in developing embryos and their subsequent effects on embryo development in vitro. The compact cumulus oocyte complexes (COCs) were cultured in tissue culture medium (TCM)-199 supplemented with 10 ng/ml epidermal growth factor, 4 IU/ml pregnant mare serum gonadotropin (PMSG), and human chorionic gonadotropin (hCG) and 10% (v/v) porcine follicular fluid (pFF) for 44 h. After maturation culture, the denuded oocytes were activated with a single DC pulse of 2.0 kV/cm in 0.3 M mannitol solution containing 0.5 mM of HEPES, 0.1 mM of $CaCl_2$ and 0.1 mM of $MgCl_2$ for $30{\mu}s$ using a BTX Electro-cell Manipulator. The activated oocytes were cultured in modified North Carolina State University-23 (mNSCU-23) medium for 168 h. The level of $H_2O_2$ in each embryo was measured by the dichlorohydrofluorescein diacetate (DCHFDA) method at 48 h after activation. The blastocyst formation rate was significantly higher when culture medium was supplemented with 50 and $100{\mu}M$ L-ascorbic acid (31.2 and 38.7%, respectively) compared to non-supplemented (16.1%) group. Accordingly, significantly more cells in blastocyst were found for 50 and $100{\mu}M$ L-ascorbic acid (50.0 and 56.4, respectively) compared to 0 and $200{\mu}M$ L-ascorbic acid (36.5 and 39.8, respectively). L-ascorbic acid reduces the $H_2O_2$ level in developing embryos in a dose-dependant manner. The $H_2O_2$ level (pixels/ embryos) was 191.5, 141.0, 124.0 and 163.3 for 0, 50, 100 and $200{\mu}M$ L-ascorbic acid, respectively. So, we recommend to supplement 50 or $100{\mu}M$ L-ascorbic acid in porcine in vitro culture medium.

• Journal of Aquaculture
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• v.18 no.2
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• pp.115-121
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• 2005

This study was performed to investigate the formation and mass production of resting eggs by freshwater rotifer, Brachionus calyciflorus as influenced by different aeration supplements and exchange intervals of culture water in 15-L culture vessels and 500 L culture tanks. The maximum densities and mixis rates of the rotifers were not different between experimental group exposed to air or oxygen supplements. However, the fertilization rate and formation of resting eggs of the rotifers in the air-supplemented group were significantly higher than those in the oxygen supplemented group. In the experiment concerning exchange interval of culture water, the maximum density of the rotifers and formation of resting eggs in the batch culture were significantly higher than those in the semi-continuous culture with exchange of water every day. The formation of resting eggs per Chlorella dry weight was highest in the semi-continuous culture with exchange of water every day. The resting eggs of rotifers were produced at a density of $51.8{\sim}57.9{\times}10^6$ eggs in 500-L culture tanks. In this study, the batch culture with air is an effective method for mass production of resting eggs by the freshwater rotifer, B. calyciflorus, and the efficiency of mass production of resting eggs by this rotifer was similar to that of the seawater rotifers, B. plicatilis and B. rotundiformis.

• Korean Journal of Community Nutrition
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• v.13 no.2
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• pp.276-287
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• 2008

Diabetic mellitus in an older population is associated with increased basal oxidative stress and free radical accentuated by hyperglycemic challenge. Enhanced free radical in diabetic elderly can cause the oxidative damage and such damage can be protected by antioxidant defense system. It is believed that vitamin C, A and E are the most abundant and effective antioxidants in human plasma. The purpose of this study was to determine the antioxidant status in Korean diabetic elderly using the case-control study. The antioxidant status was examined by determining plasma levels of antioxidant vitamins (vitamin C, A, E, ${\beta}$-carotene), total antioxidant status (TAS) and thiobarbituric acid reactive substance (TBARS) and intakes of vitamin C, A, ${\beta}$-carotene and retiol. Fasting glucose and HbA1c levels and serum lipid profiles (triglyceride (TG), total cholesterol, HDL-cholesterol and LDL-cholesterol) were also determined. Diabetic subjects were 122 elderly persons over 60 years old, visiting public health center, and control subjects were 96 healthy elderly persons living in Ulsan, Korea and they were matched by age, gender, smoking and drinking status. The diabetic and control subjects were divided into sub-groups according to the status of using diet therapy and vitamin supplement. The subjects were interviewed to collect data on their general characteristics, disease history, vitamin supplement, diet therapy and health-related habits by questionnaires. Their dietary intakes were obtained by means of semi-quantitative food frequency questionnaires (SQFFQ). Fasting plasma glucose and HbA1c levels were significantly higher in diabetes than in control subjects, and plasma total cholesterol level of diabetes was not significantly different from that of control subjects. However serum HDL cholesterol level of diabetes was significantly lower and serum TG level of diabetes was significantly higher than those of control group. The average vitamin A and ${\beta}$-carotene intakes of diabetes were significantly higher than those of control subjects. There was no significant difference in plasma vitamin C, ${\beta}$-carotene, and TBARS levels between two groups, but plasma vitamin A, E and TAS levels were significantly higher in diabetes than those in control group. Plasma vitamin A and TAS levels of diabetic subjects using diet therapy were higher than those of control using diet therapy, and plasma vitamin E, ${\beta}$-carotene and TAS levels of diabetic subjects using vitamin supplements were significantly higher than those of controls using vitamin supplements. These results suggested that diabetic mellitus could enhance antioxidant defences against reactive oxygen species and interest in healthy eating such as consumption of more antioxidant nutrients.

• Journal of Chest Surgery
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• v.29 no.6
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• pp.593-598
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• 1996

The large number of past investigation on extended myocardial protection clearly indicates that cold potassium cardioplegia and topical cooling have limited capabilities. Accordingly, more recent experimen- tal approaches have focused on the modalities of reperfusion and their implication on postischemic myo- cardial recovery. Oxygen may play a crucial role in the development of ischemic and reperfusion injury. Reactive oxygen radicals may be produced during ischemia or reperfusion after incomplete reduction of molecular oxygen or from other pathway and then induce fatal injury of the heart. The important obser- vation of oxygen-induced myocardial damage during reperfusion has led to the concept of applying oxy- gen free radical scavengers. So, this study is on dietary vitamin C supplementation as antioxidant in rats to determine whether or not they have a higher tolerance against cardiac ischemia-reperf'usion injury under Langendorff system. Male or female Sprague-Dawley rats (190-33Og) were randomly separated into two groups. Group A was not treated(n=10). Group B received vitamin C supplement (n=10). Experiment was performed 24 hours after vitamin C 200mg fed orally as injectable ascorbic acid. There were significant differences in contractile parameters between control and vitamin C-treated group. The RLVP (r te of post/preischemic left ventricular pressure) and Rdp/dt (rate of post/preischemic dp/dt) were significant statistically between two groups (p<0.05). But, RHR (rate of post/preischemic heart rate), time to first beat and sta'utilization were not significant. In conclusion, pretreatment with the antioxidant, ascorbic acid, was found to preserve left ventricular contractile function. But the precise mechanism of action of ascorbic acid has not as yet been determined, so further study will be required.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.2
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• pp.244-249
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• 2003

To investigate the hepatic oxygen free radical metabolizing system and changes of serum cholesterol levels in rats fed a diet supplemented with Monascus pigment (MP), Sprague-Dawley rats weighing about 300 g have been fed a diet supplemented with 2% or 4% MP for a month. The rats fed 2% MP supplemented diet gained less body weight than the control rats and those fed 4% W supplemented diet. Those fed 2% or 4% MP supplemented diet had no remarkable changes in liver function on basis of liver weight/body weight, serum levels of xanthine oxidase, alanine amino transferase activity In rats fed 2% and 4% MP supplemented diet, hepatic cytochrome P45O dependent aniline hydroxylase activity significantly (p<0.05) declined about 32%, 37% respectively and showed no significant differences between rats fed 2% and 4% MP supplemented diet whereas those fed 2% MP supplemented diet showed about 29% increased hepatic xanthine oxidase activity. And hepatic glutathione S-transferase and glutathione peroxidase activites in rats fed 2% MP supplemented were more increased by about 17%, 28% respectively than the control rats. There were no significant differences both in between those fed 2% and 4% MP supplemented diet. Especially rats fed 2% or 4% MP supplemented diet showed a significant (p<0.05) increase in hepatic catalase activity by 41%, 25% compared with control rats and those fed 4% MP supplemented diet showed more decrease in tendency of catalase activity than those 2% MP supplemented diet. But hepatic superoxide dismutase activity and glutathione content were appeared to be similar value among three groups. On the other hand, rats fed 2% MP supplement diet showed 17% increased levels of serum HDL-choresterol and 26% decreased value of LDL-cholesterol and serum level of triglyceride. But no different value were appeared between those fed 2% and 4% MP supplemented diet. Especially in those fed 2% and 4% MP supplemented diet, artherogenic index were significantly (p<0.05) declined by 37%, 29% respectively compared with control. In conclusion, it is likely that rats fed a diet supplemented with a proper quantity of MP may have the potential of oxygen free radical detoxication and lowering of artherogenic index.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.748-752
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• 1999

This study was carried out to compare the growth and fatty acids composition of the rotifer (Brachionus rotundiformis) cultured in high density by the various enrichments and culture methods. The rotifer fed on condensed freshwater Chlorella was enriched with $\omega$-yeast, Algamac, Super Selco and marine Chlorella. In another culture method, the rotifer was cultured with enrichment supplements for 6 hours after feeding with condensed freshwater Chlorella supplement for 18 hour. The rotifer fed with condensed marine Chlorella for 24 hours without fieshwater Chlorella was used as a control group. Culture tanks (5 $\ell$ working volume) was immersed in a water bath ($28^{\circ}C$). The density of rotifer and dissolved oxygen level in water was stable in control group of rotifer cultured with condensed marine Chlorella for 24 hours and the n-3 HUFA content of rotifer was the highest among the rotifer culture methods. However, the density of rotifer and dissolved oxygen level in the groups of rotifers enriched with $\omega$-yeast, Algamac and Super Selco by methods were drastically decreased, The n-3 HUFA contents of rotifers enriched by Super Selco were higher than those of rotifer enriched by either $\omega$-yeast or Algamac in both methods. The results from this experiment indicated that supplementation of condensed marine Chlorella for 24 hour by the semi-continuous culture was effective for the improvement of the nutritional value of rotifer and it could provide the stable growth condition for rotifer culture in high density.

• The Journal of Korean Medicine
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• v.25 no.2
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• pp.127-137
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• 2004

Objectives : The purpose of this investigation was to evaluate the effects of Sohaphyang-won (SH) on the alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E18 rat cortical cells were grown in neurobasal medium containing B27 supplement. On 12 DIV, SH was added ($20\mu\textrm{g}/ml$) to the culture media for 24 hrs. On 14 DIV, cells were given a hypoxic insult (2% O2/5% CO2, $37^{\circ}C$, 3 hrs), returned to normoxia and cultured for another 24 hrs. Total RNA was prepared from SH-untreated (control) and -treated cultures and alteration in gene expression was analyzed by microarray using rat 5K-TwinChips. Results : Effects on some of the genes whose functions are implicated in neural viability are as follows: 1) For most of the genes altered in expression, the global M values were between -05 to +0.5, Among these, 1517 genes were increased in their expression by more than global M +0.1, while 1480 genes were decreased by more than global M -0.1. 2) The expression of apoptosis-related genes such as Bad (global M =0.35), tumor protein p53 (T53) (global M =0.28) were increased, while v-akt murine thymoma viral oncogene homolog 1 (Akt1) was decreased. 3) The expression of hemoglobin alpha 1 (probably neuroglobin) was increased by about 3.2-fold (global M =1.7). 4) The expression of antioxidation-related catalase gene was increased (global M =0.26). 5) The expression of PKCzeta (prkcz), an upstream kinase of MAPK, was increased (global M =0.29). 6) The expression of retinoic acid receptor alpha (RAR), which may regulate transcription in hypoxic stress, was increased (global M =10.27). Conclusions : In summary, the microarray data suggest that SH doesn't increase the expression of oxygen capture-, anti-oxidation- and 'response to stress' -related genes but decreases some anti-apoptosis genes which would help protect the hypoxic cells from apoptosis.

• The Journal of Internal Korean Medicine
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• v.34 no.1
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• pp.100-111
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• 2013

Objectives : This study investigated the impact of Caesalpinia sappan L. on oxidative damage and inflammatory relevant factor in RAW 264.7 cells and human umbilical vein endothelial cells (HUVEC). Methods : We determined whether fractionated EtOH extracts of Caesalpinia sappan L. (CSL) inhibit free radical generation such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), reactive oxygen species (ROS) and nitric oxide (NO) and pro-inflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 cells and HUVEC. Result : 1. DPPH removal capacity was increased by CSL. 2. LPS-induced ROS, and NO inhibitory capacity were increased by CSL. 3. LPS-induced cell death of Raw 264.7 cells was decreased by CSL. 4. The amount of cytokine generation in Raw 264.7 cell was decreased significantly by CSL. 5. The amount of cytokine generation in HUVEC was decreased significantly by CSL. Conclusions : These results suggest that CSL supplement may attenuate oxidative stress by elevated antioxidative processes, and suppress inflammatory mediator activation.

• Korean Chemical Engineering Research
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• v.51 no.6
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• pp.727-732
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• 2013

In this report, we present an inverse opal scaffold that can enhance the chondrogenic differentiation of human adipose-derived stem cells (hADSCs) without drug, gene, or cytokine supplement. Inverse opal scaffolds based on poly(D,L-lactide-co-glycolide) were formed with uniform $200{\mu}m$ pores. Due to uniform pore sizes and well-controlled interconnectivity of inverse opal scaffold, hADSCs were allowed to distribute homogeneously throughout the scaffolds. As a result, high cell density culture with scaffold was possible. Since the hADSCs cultured in inverse opal scaffolds were subjected to limited supplies of oxygen and nutrients, these cells were naturally preconditioned to a hypoxic environment that stimulated the up-regulation of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). As a result, apoptotic activity of hADSCs until 3 weeks after initial cell seeding was significantly reduced and chondrogenic differentiation related molecular signal cascades were up regulated (transforming growth factor-beta, phosphorylated AKT, and phosphorylated p38 expression). In contrast, hADSCs cultured with small and non-uniform porous scaffolds showed significantly increased apoptotic activity with decreased chondrogenic differentiation. Taken together, inverse opal scaffold could potentially be used as an effective tool for improving chondrogenesis using stem cells.