• 대한생식의학회:학술대회논문집
• /
• 대한불임학회 2002년도 제42차 춘계학술대회
• /
• pp.91.2-92
• /
• 2002

• Reproductive and Developmental Biology
• /
• 제35권2호
• /
• pp.191-195
• /
• 2011

The present study was conducted to examine the reactive oxygen species (ROS) generation levels in porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by the combination of electric stimulus and 6-DMAP before in vitro culture. Porcine oocytes and parthenogenetic embryos were stained in 10 ${\mu}M$ dichlorohydrofluorescein diacetate (DCF) or 10 ${\mu}M$ hydroxyphenyl fluorescein (HPF) dye each for 30 min at $39^{\circ}C$. The fluorescent emissions from the samples were recoded as JPEG file and the intensity of fluorescence in oocytes and embryos were analyzed. $H_2O_2$ and $^{\cdot}OH$ radical levels of porcine oocytes were reduced immediately after electric stimulation. However, $H_2O_2$ and $^{\cdot}OH$ radical levels of parthenogenetic embryos were increased with time elapsed after electric stimulation from 0 h to 3 h and after DMAP culture. During in vitro culture, $H_2O_2$ and $^{\cdot}OH$ radical levels were gradually increased from the one-cell stage to the two- and four-cell stages. The result of the present study suggests that the ROS was not increased by electric pulse in porcine embryos. Rather than it seems to be associated with the stage of development and the culture condition.

• Ocean and Polar Research
• /
• 제29권2호
• /
• pp.187-191
• /
• 2007

In this study, antioxidant activities of acetone/dichloromethane and methanol extracts of Arctic seaweeds were investigated. The antioxidant properties of both extracts of arctic seaweed were evaluated using two different peroxynitrite tests, including scavenging power on authentic peroxynitrite and inhibitory activities on peroxynitrite generation from 3-morpholinosydnonimine (SIN-1) producing superoxide anion and nitric oxide simultaneously in vitro. At concentration of $10\;{\mu}g/ml$, the acetone/dichloromethane and methanol extracts of Odonthalia dentata exhibited 54.6 and 64.2% inhibition against peroxynitrite generation from SIN-1 while they exhibited 24.3 and 23.0% scavenging activities on authentic peroxynitrite, respectively. On the other hand, the acetone/dichloromethane extract of Polysiphonia stricta exhibited 61.8% inhibition only against the induced peroxynitrite from SIN-1. Furthermore, the crude extracts of Odonthalia dentata and Polysiphonia stricta were fractionated into n-hexane, 85% aq. MeOH, n-BuOH, and $H_2O$ fractions, successively, and only 85% aq. MeOH fraction exhibited the best inhibition.

• Reproductive and Developmental Biology
• /
• 제37권1호
• /
• pp.51-55
• /
• 2013

The present study was conducted to examine the effect of antioxidant treatment during parthenogenetic activation procedure on the reactive oxygen species (ROS) levels and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by a combination of electric stimulus and 2 mM 6-dimethylaminopurine (6-DAMP) before in vitro culture. During the activation period, oocytes were treated with $50{\mu}M$ ${\beta}$-mercaptoethanol (${\beta}$-ME), $100{\mu}M$ L-ascorbic acid (Vit. C) or $100{\mu}M$ L-glutathione (GSH). To examine the ROS level, porcine parthenogenetic embryos were stained in $10{\mu}M$ dichlorohydrofluorescein diacetate ($H_2DCFDA$) dye 20 h after culture, examined under a fluorescence microscope, and the fluorescence intensity (pixels) were analyzed in each embryo. The parthenogenetic embryos were cultured for 6 days to evaluate the in vitro development. The apoptosis was measured by TUNEL assay. The $H_2O_2$ levels of parthenogenetic embryos were significantly lower in antioxidant treatment groups ($26.9{\pm}1.6{\sim}29.1{\pm}1.3$ pixels/embryo, p<0.05) compared to control ($33.2{\pm}1.7$ pixels/embryo). The development rate to the blastocyst stage was increased in antioxidant treatment groups (32.0~32.5%) compared to control (26.9%, p<0.05), although, there was no difference in apoptosis among groups. The result suggests that antioxidant treatment during parthenogenetic activation procedure can inhibit the ROS generation and enhance the in vitro development of porcine parthenogenetic embryos.

• 한국발생생물학회지:발생과생식
• /
• 제22권3호
• /
• pp.235-244
• /
• 2018

We investigate the effect of endoplasmic reticulum (ER) stress inhibitor treatment during parthenogenetic activation of oocytes on the ER stress generation, apoptosis, and in vitro development of parthenogenetic porcine embryos. Porcine in vitro matured oocytes were activated by 1) electric stimulus (E) or 2) $E+10{\mu}M$ Ca-ionophore (A23187) treatment (EC). Oocytes were then treated by ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxychloic acid (TUDCA, $100{\mu}M$) for 3 h prior to in vitro culture. Parthenogenetic embryos were sampled to analyze ER stress and apoptosis at the 1-cell and blastocyst stages. The x-box binding protein 1 (Xbp1) mRNA and ER stress-associated genes were analyzed by RT-PCR or RT-qPCR. Apoptotic gene expression was analyzed by RT-PCR. At the 1-cell stage, although no difference was observed in Xbp1 splicing among treatments, BiP transcription level in the E group was significantly reduced by salubrinal treatment, and GRP94 and ATF4 transcription levels in EC group were significantly reduced by all treatments (p<0.05) compared to control. In the EC group, both apoptotic genes were reduced by ER stress inhibitor treatments compared to control (p<0.05) except Caspase-3 gene by TUDCA treatment. These results suggest that the treatment of ER stress inhibitor during parthenogenetic activation can reduce ER stress, and thereby reduce apoptosis and promote in vitro development of porcine parthenogenetic embryos.

• Journal of Ginseng Research
• /
• 제21권2호
• /
• pp.78-84
• /
• 1997

As a part of our ongoing effort to develop new antineoplastic immunostimulator from natural sources, bioassay-directed fractionationn of polysaccharides from Korean red ginseng was carried out by observing the proliferation of marine spleen cells and the generation of lymphoklne activated killer (LAK) cells. The acidic polysaccharide fractions proliferated spleen cells and generated LAK cells in proportion to their acidity in vitro. The LAK cell which was induced by ginseng showed tumoricidal activity against both NK celt sensitive and insensitive tumor target cells without major histocompatibility (MHC) restriction. Adherent macrophages and CD4+helper T cells were involved in the generation of the LAK cells. The acidic polysaccharide from Korean red ginseng synerglzed with recombinant IL-2 (rIff-2) at lower than 3 U/ml. The optimal doses of the acidic polysaccharide from Korean red ginseng for the proliferation of spleen cells and for the generation of LAK cells were 1 mg/ml and 100 $\mu\textrm{g}$/ml, respectively; this means that the mechanisms for the both activities may be different from each other.

• Journal of Ginseng Research
• /
• 제41권3호
• /
• pp.233-239
• /
• 2017

Background: Nephrotoxicity is the major side effect in cisplatin chemotherapy. Previously, we reported that the ginsenosides Rk3 and Rh4 reduced cisplatin toxicity on porcine renal proximal epithelial tubular cells (LLC-PK1). Here, we aimed to evaluate the protective effect of ginsenosides Rk3 and Rh4 on kidney function and elucidate their antioxidant effect using in vitro and in vivo models of cisplatin-induced acute renal failure. Methods: An enriched mixture of ginsenosides Rk3 and Rh4 (KG-KH; 49.3% and 43.1%, respectively) was purified from sun ginseng (heat processed Panax ginseng). Cytotoxicity was induced by treatment of $20{\mu}M$ cisplatin to LLC-PK1 cells and rat model of acute renal failure was generated by single intraperitoneal injection of 5 mg/kg cisplatin. Protective effects were assessed by determining cell viability, reactive oxygen species generation, blood urea nitrogen, serum creatinine, antioxidant enzyme activity, and histopathological examination. Results: The in vitro assay demonstrated that KG-KH ($50{\mu}g/mL$) significantly increased cell viability (4.6-fold), superoxide dismutase activity (2.8-fold), and glutathione reductase activity (1.5-fold), but reduced reactive oxygen species generation (56%) compared to cisplatin control cells. KG-KH (6 mg/kg, per os) also significantly inhibited renal edema (87% kidney index) and dysfunction (71.4% blood urea nitrogen, 67.4% creatinine) compared to cisplatin control rats. Of note, KG-KH significantly recovered the kidney levels of catalase (1.2-fold) and superoxide dismutase (1.5-fold). Conclusion: Considering the oxidative injury as an early trigger of cisplatin nephrotoxicity, our findings suggest that ginsenosides Rk3 and Rh4 protect the kidney from cisplatin-induced oxidative injury and help to recover renal function by restoring intrinsic antioxidant defenses.

• Archives of Pharmacal Research
• /
• 제29권6호
• /
• pp.469-475
• /
• 2006

A series of yakuchinone B 1f and its analogs 1a-e was synthesized and evaluated for free radical scavenging, suppression of LPS-induced NO generation, cytotoxicity and anti-excitotoxicity in vitro. Compound 1c exhibited potent anti-excitotoxicity, while all compounds 1a-f showed considerable effects of free radical scavenging, suppression of LPS-induced NO generation, and cytotoxicity in microglia.

• Journal of Plant Biotechnology
• /
• 제1권1호
• /
• pp.8-12
• /
• 1999

New winter wheat winter barley hybrids were produced (Mv9 kr1 Igri, Mv9 kr1 Osnova, Asakazekomugi Manas). The wheat-barley hybrids showed entire male sterility and were multiplied in tissue culture. Chromosome configurations were studied with GISH in meiosis in the Mv9 krl x Igri hybrid and in its progenies multiplied in vitro. Chromosome pairing between wheat and barley has been observed in some cells in the hybrids multiplied in vitro. Backcross plants with 43 and 44 chromosomes were selected with the aim of developing new wheat-barley addition lines. Wheat-barley translocations were demonstrated with GISH in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) x barley (Hordeum vulgare L. cv. Betzes) hybrids. Five different translocations were observed. Sequential N-banding and GISH analyses were performed to further identify the translocations. The N-banding pattern of the Robertsonian translocation suggests that this chromosome consists of the short arm of barley chromosome 4H translocated to the long arm of 2B of wheat. Plants with four different homozygous translocations were selected from the following BC2F3 generation.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2019년도 추계학술대회
• /
• pp.16-16
• /
• 2019

The Cassava (Manihot esculenta Crantz) is a perennial woody shrub cultivated mainly in the tropics for its starchy tuberous roots. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. Thus, a study was undertaken to develop an efficient in vitro mass propagation protocol of Manihot esculenta Crantz. Young and actively growing stem segments were excised from adult plants of cassava. Samples were cut into a 3~4 cm nodal segments with single node after sterilization, and cultivated in the different medium supplemented with various plant growth regulators for 4 weeks. For shoot multiplication, single-node stem segments, approximately 1 cm in length, were taken from in vitro derived shoots and subcultured. After 4~6 weeks, the shoot generation rate was 55.6%, the shoot number and its length were 1.0/explant and 2.3 cm in the most favorable medium composition. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.