• Clinical and Experimental Pediatrics
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• v.52 no.2
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• pp.213-219
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• 2009

Purpose:Iron is a critical nutritional element that is essential for a variety of important biological processes, including cell growth and differentiation, electron transfer reactions, and oxygen transport, activation, and detoxification. Iron is also required for neoplastic cell growth due to its catalytic effects on the formation of hydroxyl radicals, suppression of host defense cell activities, and promotion of cancer cell multiplication. Chronic transfusion-dependent patients receiving chemotherapy may have iron overload, which requires iron-chelating therapy. We performed this study to demonstrate whether the iron chelating agent deferoxamine induces apoptosis in Saos-2 osteosarcoma cells, and to investigate the underlying apoptotic mechanism. Methods:To analyze the apoptotic effects of an iron chelator, cultured Saos-2 cells were treated with deferoxamine. We analyzed cell survival by trypan blue and crystal violet analysis, apoptosis by nuclear condensation, DNA fragmentation, and cell cycle analysis, and the expression of apoptotic related proteins by Western immunoblot analysis. Results:Deferoxamine inhibited the growth of Saos-2 cell in a time- and dose-dependent manner. The major mechanism for growth inhibition with the deferoxamine treatment was by the induction of apoptosis, which was supported by nuclear staining, DNA fragmentation analysis, and flow cytometric analysis. Furthermore, bcl-2 expression decreased, while bax, caspase-3, caspase-9, and PARP expression increased in Saos-2 cells treated with deferoxamine. Conclusion:These results demonstrated that the iron chelating agent deferoxamine induced growth inhibition and mitochondrial-dependent apoptosis in osteosarcoma Saos-2 cells, suggesting that iron chelating agents used in controlling neoplastic cell fate can be potentially developed as an adjuvant agent enhancing the anti-tumor effect for the treatment of osteosarcoma.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.51-61
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• 2001

This study was conducted to determine developmental ability of reconstructed embryos by nuclear transfer using somatic cell of Korean bull with high genetic value. Fibroblast cells obtained from ear biopsy of the bull were cultured in Dulbecco's Modified Eagle's medium (DMEM) at 37$^{\circ}C$ in air containing 5% $CO_2$. The cummulus-oocyte complexes were collected from slaughterhouse and were matured in vitro for 20 h in TCM 199 culture medium and the oocytes were then enucleated in modified phosphate buffered saline with cytochalasin B. Matured bovine oocytes were enucleated by aspirating the first polar body and metaphase chromatin using a beveled pipette in modified phosphate buffered saline. The ear fibroblast cells were fused into enucleated oocyte by electrical stimulation. The reconstructed oocytes were activated with ionomycine and 6-dimethylaminopurine, and then cultured in CR1aa medium for 7.5 days. Out of 524 bovine eggs reconstructed by nuclear transfer 65.6%(277/422) embryos were cleaved, and 30.7% (85/277) cleaved embryos were developed to the morula to blastocysts. There was no difference of developmental ability in vitro of reconstructed embryos regardless of donor cell passages. In order to determine fate of foreign mitochondria of donor nucleus, the Mito Tracker stained cells were fused into enucleated oocytes. The donor mitochondria were detected early stage of embryos, but disappeared rapidly. The developmental ability of reconstructed embryos was not impaired by Mito Tracker treatments. The results indicate that viable reconstructed embryos can be producted by nuclear transfer using somatic cell of Korean bulls.bulls.

• Korean Journal of Psychosomatic Medicine
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• v.29 no.2
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• pp.207-212
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• 2021

Anti-N-methyl-D-aspartate receptor (Anti-NMDAR) encephalitis is a neuroinflammatory disease mediated by autoantibodies to NMDAR. In the initial clinical stages of anti-NMDAR encephalitis, psychiatric symptoms like delusions, perceptual disturbances, and disorganized speech or behaviors are pronounced even without obvious neurological symptoms. Early treatments like immunotherapy and/or tumor removal are central to good clinical outcomes. Hence, it is important to diagnose early anti-NMDAR encephalitis, distinguishing it from mental disorder. In the present case study, the authors described psychiatric symptoms assessed with Positive and Negative Syndrome Scale (PANSS) of Ms. A, a 26-year-old woman, in the early phase of anti-NMDAR encephalitis. We will discuss the characteristic psychopathology of anti-NMDAR encephalitis toward prompt diagnosis and treatment. Ms. A showed a higher negative subscale score than positive one on the PANSS. Compared with mental disorder, negative symptoms and cognitive impairment would be more prominent in the early stage of anti-NMDAR encephalitis. Rituximab and teratoma removal were effective, and quetiapine showed good tolerability. It is recommended to evaluate anti-NMDAR encephalitis when negative symptoms, cognitive impairment, catatonia, changes in consciousness level, and neurological symptoms are observed, especially in young women.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.155-161
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• 2000

The morphological changes on nuclear phase of the germinal vesicle of porcine follicular oocytes during in vitro culture were examined. The high rates (75~77%) of the oocytes collected from follicles of 1~2mm or 6~100mm in diameter were at the GV-I to GV-II stages. When oocytes with or without cumulus cells after collection from follicles of 2~6mm in diameter were cultured for 5 h, the rates of oocytes at GV-IV to GV-Ⅵ stages were higher in oocytes with (52%) than in oocytes without (30%) cumulus cells. After 1 h of oocyte culture, there was no differences in the distribution of GV-IV to GV - Ⅵ stages in the media with or without catalase, xanthine and catalase＋xanthine. After 5 h of culture, however, the distribution of GV-IV to GV-Ⅵ stages were 46, 69, 69 and 70% for medium with none, catalase, xanthine and catalase＋xanthine. The highest rate of GVBD was also observed in the medium with catalase＋xanthine (6%). These results indicate that exposure of porcine follicular oocytes to catalase＋xanthine excels maturation to GV stage and enhances oocyte nuclear maturation.

• Journal of Veterinary Clinics
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• v.31 no.1
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• pp.25-30
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• 2014

This study tested the hypothesis that the priming effects of progesterone on the timing of the LH surge induced by exogenous estradiol are more potentiated the negative feedback actions of progesterone on LH secretion by the existence of estradiol. In previous studies, the time interval from estradiol infusion until the peak of LH surge was gradually and significantly extended by the different levels of progesterone treated before estradiol infusions. Longterm ovariectomized Shiba goats that had received implants of estradiol capsules (Day 0) and three different progesterone silastic packet inducing follicular, subluteal and luteal levels of progesterone were divided into three groups such as non-P, low-P and high-P group. Blood samples were collected daily throughout the experiment for the analysis of gonadal steroid hormone levels. On Day 7, all devices of progesterone and estradiol packets were removed but estradiol capsules were maintained during the experiment, and blood samples were collected at 1 hr interval for 12 h from the time of progesterone removals to determine peripheral changes of estradiol and progesterone concentration. Then all animals were infused estradiol on the Day 7 after 13 h from the removals of progesterone devices with a peristaltic pump into jugular vein at a rate of 3-6 ${\mu}g/h$ for 36 h. For analysis of peripheral LH and estradiol concentration, blood samples were collected via another jugular vein at 2 h intervals for 52 h (from 4 h before the start of estradiol infusion to 48 h after the start of estradiol infusion). In all animals of the three groups treated with estradiol infusion, an LH surge was expressed but the peak time of LH surge was different. This time interval was not extended by the different levels of progesterone treated before estradiol infusions and the difference was not significant during this interval between the Low P and the High P groups. Progesterone pretreatment may contribute to regulating the neural system that is responded by estradiol, and estradiol existence potentiates the negative feedback effect of progesterone on GnRH/LH surge-generating system.

• Development and Reproduction
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• v.6 no.2
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• pp.117-122
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• 2002

Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX＋Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX＋Oil group and in OVX＋E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX＋Oil and OVX＋E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.

• Development and Reproduction
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• v.5 no.2
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• pp.175-180
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• 2001

The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E$_2$) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E$_2$ significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.

• Korean journal of applied entomology
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• v.36 no.1
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• pp.88-95
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• 1997

Olfactometer was used to find the attractiveness of kairomone extracted from gypsy moth (Lymantria dispar) to the egg parasitoid (Ooencyrtus kuvanae). The attractiveness of kairomone extracted from scales and hairs which were attached to the surface of the eggs were higher than that of whole eggs with 79.9% and 67.6% respectively. The probing lantency of the egg parasitoid on the extract of scales and hairs were 115.5 sec. and that of whole eggs were 113.2 sec. Eggs taken from egg mass were shown 91.0% of attractiveness to the egg parasitoid, however only 15% of egg parasitoid were attracted by follicles dissected from the host ovary. The probing latency of egg mass was faster than that of follicles dissected from the host with 84.2 sec. and 114.0 sec. respectively. Egg parasitoid showed the most active attractiveness (88.0%), relatively longer examing period (89.2 sec.) and shorter probing latency (26.8 sec.) to the adhesive materials secreted from reproductive accessory glands of the host. Out of six organic solvents used for the extraction of kairomone from the host egg mass, n-hexane fraction received the most active response from xthe egg parasitoid and the parasitoid reared in vivo showed 3 times higher attractiveness to the n-hexane fraction than that of in vitro.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.127-127
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• 2003

본 연구의 목적은 젖소에서 다배란유기법의 확립을 위해서 호르몬을 처리법을 달리한 결과이다. 난포의 발생을 자극하는 FSH 호르몬을 하루에 2번의 호르몬 주사를 하는 데, 이는 혈액내에 난포자극호르몬의 농도를 연속적으로 유지하여 난소에서 다난포 발생의 목적을 달성하고자 하는 방법이다. 하지만 빈번하게 호르몬을 주사하는 방법은 처리하는 노력이 필요하고 공란우도 스트레스로 작용한다. 이러한 단점을 극복하기 위해 한번에 많은 난포의 성숙을 유도하는 다배란처리법에 대한 보고가 많이 있어, 본 연구에서 이 다배란처리 효과를 비교하고자 젖소를 이용하였다. 방법으로는 다회주사법(8회)과 일회주사법(1회)에서 황체수와 수정란의 생산수 등을 조사하였다. 다회주사법은 생리식염수에 융해한 FSH를 1일 50mg씩 2회(12시간 간격)주사하는 법으로 CIDR 질내 주입 9일째부터 4일간 주사하였으며, 질내에 CIDR를 주입하고 11일째되는 날에 PG(25mg, 루텔라이스, 한국)를 주사하고, 12일째는 CIDR를 제거하여 일회주사법과 주기를 맞추었다. 일회주사법은 CIDR를 질내에 주입하고 나서 9일째 되는 날에 400mg의 FSH을 생리식염수에 융해하여 일회주사를 하고 다회주사법과 같이 인공수정과 채란시간을 맞추었다. 인공수정은 황체퇴행제(PG)를 주사하고 나서 48, 72시간에 2회 인공수정을 실시하여 1주일 후 수정란을 채란하였다. 인공수정시는 반드시 GnRH(2.5mg)를 동시에 주사하였다. 채란은 인공수정 후 7일째 되는 날에 비외과적으로 채란을 실시하여 우수한 수정란은 동결을 실시하였다. 다회주사법에서 14두, 일회주사법에서 14두를 공시하여 황체수, 회수수정란수, 배반포수, 동결란수를 조사하였다. 다회주사법에서는 황체수는 8.07$\pm$6.62개, 회수정란수는 6.78$\pm$5.96개, 배반포율은 58.9%, 동결란 생산율은 52.6%의 성적을 얻었으며, 일회주사법에서 황체수는 12.07$\pm$8.07개, 회수수정란수는 10.0$\pm$9.24개, 배반포율은 27.1%, 동결란 생산율은 25.0% 수준의 결과를 얻었으나 결과에 대한 통계분석시 다회주사법과 일회주사리법 간에 유의적인 차는 인정되지 않았다(student T-test, P<0.05). 본 실험의 결과에서 나타나듯이 생리식염수를 용매로 이용한 FSH 1회 주사법이 다회주사법과 비교하였을 때에 수정란 생산효율에서 유의적인 차이가 없으므로 이 방법을 이용한 다배란유기는 노동력과 공란우에 대한 스트레스를 경감할 수 있을 것으로 사료된다.

• Development and Reproduction
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• v.3 no.1
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• pp.101-105
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• 1999

N-Methylpurine-DNA glycosylase (MPG) removes N-methylpurine and other damaged purines in DNA. RT-PCR analysis revealed MPG mRNA expression at various tissues of fetal development from day 8 to day 18 fetus and day 400 mature adult. The MPG transcripts were abundant during fetal development in mice. In placenta, the MPG mRNA was continuously decreased from day 8 post coitum (p.c) to day 18 p.c. fetus. The high level of mRNA in fetal brain and liver was drastically declined in day 400 mature adult. The expression of MPG, originally characterized by its highest level of expression in the epididymis of adult mouse, was detected with high level in several other reproductive organ, including the ovary, oviduct, testis, vas deference, uterus, and seminal vesicles. These results demonstrate developmental stage- and tissue-specific variation of MPG gene expression.