2000년부터 시작된 6·25 전사자 유해발굴사업은 전쟁 기간 동안 수 습되지 못한 전사자 13만여 분을 찾아 국가와 가족의 품으로 모신다는 데 목적을 두고 추진 중이다. 2005년부터 국방부 유해발굴감식단(이하 국유단)을 출범하여 유전자 채취와 신원확인센터 설치 등 체계적 및 전문적으로 추진하고 있는 영구적인 국가정책 사업이다. 아울러 2018 년 4·27 판문점 선언과 9.19 남북군사합의에 의해서 오랜 숙원 사업으 로 제기됐던 비무장지역(DMZ) 내 유해발굴이 가능해졌다. 오랜 기간 접근하지 못했던 DMZ 내에서 남북유해발굴 공동합의는 접경지역과 한 반도에서의 군사적 긴장완화에 대한 남북한의 의지를 잘 반영한 것이 다. 기대했던 북한의 약속 이행은 없었지만 국방부는 2019년 4월 1일 부터 2021년 6월 24일까지 약 2년 반 동안 “9·19 남북군사합의”에 명 시된 남북공동유해발굴의 사전준비 차원에서 화살머리고지 일대 남쪽 지역에서 유해발굴을 꾸준히 진행했다. 특히 DMZ 내 화살머리고지 유 해발굴을 통해 9명의 국군전사자의 신원을 확인하여 유해봉환 및 안장 식을 거행했다. 이런 맥락에서 이 논문은 6·25 전사자 유해발굴사업의 의미와 중요성, 체계 수립을 위해서 “활동이론”(Activity theory)을 근 간으로 세부적으로 분석했다. 이를 기반으로 이 논문에서는 한반도의 지속 가능한 평화와 유지와 인도주의 차원에서의 최초의 DMZ 내 남북 공동유해발굴사업의 추진 배경과 성과, 그리고 의의를 중심으로 유해발 굴사업의 중요성과 함의에 대해 고찰했다.
Honeybees are inevitably threatened by various pathogens including Sacbrood virus (SBV) and Galleria mellonella. Recently, RNA interference (RNAi) has been suggested as a promising strategy for suppression of honey bee viruses. Also, Bacillus thuringiensis (Bt) has been widely applied for the control of lepidopteran pests such as G. mellonella. In this study, it was intended to develop dsRNA production platform using Bt. For this, the pHT1K-SBV vp1 vector which transcribes sense and anti-sense SBV vp1 gene under control of Cyt1Aa sporulation-dependent promoter was introduced into Bt strain NT0423 expressing Cry1-types toxins. SBV replication was suppressed in the worker A. cerana ingested dsRNA produced from the Bt transformant. Crystal proteins from the Bt transformant showed high level of insecticidal activity against 4th instar larvae of G. mellonella.
Sperm cryopreservation preserves genetic resources for animal breeding and for human patients who suffers from permanent testicular damage. Although the sperm cryopreservation has been used for many years, the addition of cryoprotective agent (CPA) during cryopreservation negatively affects sperm function and quality. Our previous study reported that the addition of CPA reduced bull sperm physiological functions. However, the sperm cells collected from individual bulls presented different sensitivity to the damage induced by CPA. In the present study, we examined if CPA affect sperm cells acquired from individual bulls. Individual bull spermatozoa were divided into two groups based on motility parameters; high CPA-tolerant sperm (HCS) and low CPA-tolerant sperm (LCS). Our results showed that the HCS group presented good physiological function after CPA exposure, whereas the LCS group showed a significant decrease in the sperm function. We also found differentially expressed five proteins between the HCS and LCS groups, which refer to cytosolic 5′-nucleotidase 1B (NT5C1B), fumarate hydratase (FH), F-actin-capping protein subunit beta (CAPZB), voltage-dependent anion-selective channel protein 2 (VDAC2), and cytochrome b-c1 complex subunit 1 (UQCRC1). NT5C1B and FH showed abundant expression in the HCS group, while the expression of CAPZB, VDAC2, and UQCRC1 was relatively lower in the HCS group than in the LCS group. The current results suggest that NT5C1B, FH, CAPZB, VDAC2, and UQCRC1 can be used as potential markers to predict CPA-tolerable spermatozoa. Those markers provide a reliable tool to select animals and breeds with CPA tolerance.
Recently, N-terminal pro-brain natriuretic peptide (NTproBNP) has been widely used in the areas of diagnosis, monitoring treatment efficiency, and prognosis for various heart diseases, especially heart failure (HF). In this paper, we try to estimate the prognostic significance of NT-proBNP as a risk evaluation marker in Non-ST-segment Elevation Myocardial Infarction (NSTEMI) patients. We selected NSTEMI patients who underwent percutaneous coronary intervention (PCI) primarily using a drug-eluting stent within 24 h after the onset of chest pain. We compared incidences of major adverse cardiac events (MACE) including death, myocardial infarction (MI), stent thrombosis (ST), and target vessel revascularization (TVR) in two patient groups according to a high or low serum concentration of NT-proBNP, which was measured in the emergency room (ER). We intend to minimize selection bias selecting comparing groups, considering covariate of observed variables together using propensity score matching (PSM) and propensity score weighting (PSW) based on propensity score (PS) to control the difference in baseline characteristics between high- and low NT-proBNP groups. We found that as the log NT-proBNP value increases by 1 through a hazard function of COX’s analysis, the risk of MACE increases by 1.312 times. This result indicated that the NT-proBNP level on ER admission can be used as a significant prognostic indicator to estimate 1 year of MACE in NSTEMI patients who were treated with PCI within 24 h after the onset of chest pain.
In the present study, the effect of cysteine and NT or bisphenol A (BP) on in vitro aturation (IVM) of porcine oocytes were examined. COCs was cultured in NCSU-23 medium supplement with 10% FCS which had previously been covered with mineral oil and equilibrated in a humidified atmosphere of 5% CO2 and 95% air at 38℃. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 0.5~10.0 mM cysteine were 34.0±3.2%, 36.0±3.5%, 48.0±3.8%, 22.0±3.2%, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.5~5.0 mM NT for 48 hrs were 24.0±4.2%, 18.0±4.9%, 8.0±2.2%, respectively. NT affects oocyte in vitro maturation rate in a dose-dependent. This result were significantly lower than the control group. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM NT+5.0 mM cysteine (38.0±4.3%) were significantly higher than that of NT treatment. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.05~5.0 mM BP for 48 hrs were 20.0±4.7%, 10.0±5.3%, 6.0±3.2%, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with BP was significantly lower cultured non supplement of BP (44.0±3.5%). BP affects porcine oocyte maturation rate in a dose-dependent manner. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM BP+5.0 mM cysteine (32.0±3.2%) were increased than that of BP treatment.
Although somatic cell nuclear transfer (SCNT) has successfully been produced cloned animals in several species, the cloning efficiency is extremely low. It is generally believed that the low cloning efficiency is mainly attributed to faulty epigenetic modifications underlying the aberrant reprogramming of donor cell nuclei in recipient cytoplasm after SCNT. The nuclear reprogramming process involves epigenetic modifications, such as DNA demethylation and histone acetylation, which may be a key factor in improving the cloning efficiency. Recently, the histone deacetylase inhibitors (HDACi), such as trichosatin A (TSA) and m-carboxycinnamic acid bishydroxamide (CBHA), to increase histone acetylation have been used to improve the developmental competence of SCNT embryos. Therefore, we compared the effects of TSA with CBHA on the in vitro developmental competence and pluripotency-related gene expressions (Nanog, Oct3/4 and Sox2) in porcine cloned blastocysts. The porcine cloned embryos were treated with a 50 nM concentration of TSA or a 100 μm concentration of CBHA during the in vitro early culture (10h) after cell fusion and then were assessed to cleavage rate, development to the blastocyst stage and pluripotency-related gene expressions in NT blastocysts. All data was analyzed by chi-square. Following 4-5 replicates (245, 200 and 222 for NT, TSA and CBHA treated NT embryos respectively) there was no difference between normal NT and CBHA treated NT embryos, whereas TSA treated NT embryos was significantly decreased for cleavage rate (p<0.05). The developmental competence to the blastocyst stage in CBHA treated NT embryos (18.9%) significantly increased than that of normal NT and TSA treated NT embryos (9.4% and 11.5%) (p<0.05). In addition, all of pluripotent transcription factors (Nanog, Oct3/4 and Sox2) were highly expressed in the CBHA treated NT embryos, however, Sox2 and Oct3/4 were expressed in TSA treated NT embryos and Sox2 was only expressed in normal NT embryos (p<0.05). In conclusion, the treatment of CBHA as a histone deacetylase inhibitor significantly increased the developmental competence of porcine NT embryos and pluripotency- related gene expressions (Nanog, Oct3/4 and Sox2) in NT blastocysts.
This study investigated whether the addition of porcine sperm cytosolic factor (SCF) at fusion/activation affects in vitro development of porcine parthenogenetic(PA) and nuclear transfer (NT) embryos. To determine the optimum concentration of SCF, control group of oocytes was activated with 0.3M mannitol (1.0 mM CaCl2 ․ 2H2O), other three groups of oocytes were parthenogentically activated with the fusion medium (0.1mM CaCl2 ․ 2H2O) supplemented with 100, 200 or 300 μg/ml SCF, respectively. Matured oocytes were activated with two electric pulses (DC) of 1.2 kv/cm for 30 μsec. The activated embryos were cultured in PZM-3 under 5% CO2 in air at 38.5℃ for 6 days. Oocytes activated in the presence of SCF showed a significantly higher blastocyst rate than control (p<0.05). Apoptosis rate was significantly lower in 100 μg/ml SCF group than other groups (p<0.05). Cdc2 kinase activity in control and SCF treatment group of oocytes was determined using MESACUP cdc2 kinase assay kit at 1, 5, 10, 15, 30, 45 and 60 min after activation. Cdc2 kinase activity was significantly decreased (p<0.05) in SCF group than MII oocytes or control within 5 min. For NT embryo production, reconstructed oocytes were fused in the fusion medium supplemented with 0.1 mM CaCl2 ․ 2H2O (T1), 1.0 mM CaCl2 ․ 2H2O (T2) and 0.1 mM CaCl2 ․ 2H2O with 100 μg/ml SCF (T3). Fused embryos were cultured in PZM-3 under 5% CO2 in air at 38.5℃ for 6 days. Developmental rate to blastocyst stage was significantly higher in T3 than other groups (23.0% vs. 13.5 to 15.2%) (p<0.05). Apoptosis rate was significantly lower in T3 than T1 or T2 (p<0.05). The relative abundance of Bax-α/Bcl-xl was significantly lower in in vivo or SCF group than that of control (p<0.05). Moreover, the expression of p53 and caspase3 mRNA was significantly lower in in vivo or SCF group than that of control (p<0.05). These results indicate that the addition of SCF at fusion/activation might improve in vitro development of porcine NT embryos through regulating cdc2 kinase level and expression of apoptosis related genes.
This study was conducted to investigate the efficacy of vitrification procedure for the cryopreservation of porcine oocytes and the utilization of vitrified oocytes as recipient cytoplasts for somatic cell nuclear transfer (NT), and observed that porcine oocytes are evaluated by pronuclear formation, and parthenogenetic development. Single fetal donor cells were deposited into the perivitelline space of vitrified enucleation oocytes, followed by electrical fusion and activation. NT embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at in 5% and air. 1. When the developmental rates of the oocytes after being culture for hours vitrified with EDS and ETS were 42.0%, 38.0%, respectively. This results were lower than the control group(62.2%). 2. When the developmental rates of the oocytes after being culture for hours vitrified-thawed with sucrose and glucose, 5% PVP, NCSU-23 supplemented with 10% FBS were 33.3%, 25.9%, respectively. This results were lower than the control group(55.6%). 3. The fusion and development to the blastocyst stage between the NT embryos constructed with the vitrified and non-vitrified oocytes were significant differences. Developmental rate of oocytes and NT embryos constructed with the vitrified or non-vitrified oocytes were , respectively.
In vitro development of porcine embryo is affected by culture condition. One possible factor is osmolarity of culture medium. This study examined whether high osmolarity of culture medium at the early culture stage improves development of preimplantation porcine in vitro fertilization (IVF) and nuclear transfer (NT) embryos. NT and IVF embryos were divided into three groups and the basic medium was PZM-3 (250~270 mOsmol, control group). The control group of embryos was cultured in PZM-3 for whole culture period. Other two groups of embryos were cultured in a modified PZM-3 with 0.05 M sorbitol or 0.05 M sucrose (300~320 mOsmol, sorbitol or sucrose group) for the first 2 days, and then cultured in PZM-3 for further culture. NT embryos cultured in sucrose group showed a significantly higher developmental rate to the blastocyst stage with a decreased apoptosis rate compared to the sorbitol (p<0.05). For IVF, sucrose group showed a significantly increased the blastocyst formation rate with a decreased apoptosis rate compared to the control (p<0.05). This study represents that the high osmolarity in the early embryo culture stage can enhance the in vitro development of porcine NT and IVF embryos to the blastocyst stage with reduced apoptosis of cells.
The development of neonatal neuromuscular system is accomplished by the functional interaction between the spinal neurons and its target cells, skeletal muscle cells, and the intrinsic and extrinsic factors affecting this process. The aim of this study was to identify the effect of suspension unloading (SU) and neuromuscular electrical stimulation (NMES) upon the development of the neonatal spinal cord. For this study, the neonatal rats were randomly divided into three groups: a control group, an experimental group I, and an experimental group II. The SU for experimental group I and II was applied from postnatal day (PD) 5 to PD 30, and the NMES for experimental group II was applied from PD 16 to PD 30 using NMES that gave isometric contraction with 10 Hz for 30 minutes twice a day. In order to observe the effect of SU and NMES, this study observed neutrophin-3 (NT-3) and microtubule associated protein 2 (MAP2) immunoreactivity in the lumbar spinal cord (L4-5) at the PD 15 and PD 30. The results are as follows. At PD 15, lumbar spinal cord of experimental group I and II had significantly lower NT-3 and MAP2 immunoreactivity than control group. It proved that a microgravity condition restricted the spinal development. At PD 30, lumbar spinal cord of control group and experimental group II had significantly higher NT-3 and MAP2 immunoreactivity than experimental group I. It proved that the NMES facilitated the spinal development by spinal cord-skeletal muscle interaction. These results suggest that weight bearing during the neonatal developmental period is essential for the development of neuromuscular development. Also, the NMES on its target skeletal muscle can encourage the development of the spinal cord system with a full supplementation of the effect of weight bearing, which is an essential factor in neonatal developmental process.