The aim of this study was to examine the microtubule distributions of somatic cell nuclear transfer (SCNT) and parthenogenetic porcine embryos. Porcine SCNT embryos were produced by fusion of serum-starved fetal fibroblast cells with enucleated oocytes. Reconstituted and mature oocytes were activated by electric pulses combined with 6-dimethlyaminopurine treatment. SCNT and parthenogenetic embryos were cultured in vitro for 6 days. Microtubule assembly of embryos was examined by confocal microscopy 1 hr and 20 hr after fusion or activation, respectively. The proportions of embryos developed to the blastocyst stage were 25.7% and 30.4% in SCNT and parthenogenetic embryos, respectively. The frequency of embryos showing β-tubulins was 81.8% in parthenogenetic embryos, whereas 31.3% in SCNT embryos 1 hr after activation or fusion. The frequency of the embryos underwent normal mitotic phase was low in SCNT embryos (40.6%) compared to that of parthenogenetic ones (59.7%) 20 hr after fusion or activation (p<0.05). The rate of SCNT embryos with an abnormal mitosis pattern is about twice compared to that of parthenogenetic ones. The spindle assembly and its distribution of SCNT embryos in the first mitotic phase were not different from those of parthenogenetic ones. The result shows that although microtubule distribution of porcine SCNT embryos shortly after fusion is different from parthenogenetic embryos, and the frequency of abnormal mitosis 20 hr after fusion or activation is slightly increased in SCNT embryos, microtubule distributions at the first mitotic phase are similar in both SCNT and parthenogenetic embryos.

This study was conducted to examine the protein kinase inhibitors, 6-dimethylaminopurine (DMAP) and cycloheximide (CHXM) on the development and chromosome constitution of porcine parthenogenetic embryos. In vitro matured oocytes were activated by electric stimuli (ES) or a combination of ES with culture in 2 mM DMAP or 10 μg/ml CHXM for 4 hr. Activated oocytes were cultured in PZM-3 for 6 days. Some 1-cell embryos and blastocysts were fixed by air dry method to analyze the chromosome constitutions and/or total cell number. Blastocyst development of DMAP-treated group (26.7%) was significantly higher (p<0.05) than those of CHXM-treated and ES control groups. Ploidy in 1-cell stage embryos was not different among groups (77.3 to 81.0%), however, proportion of diploid chromosome constitutions was high in DMAP-treated group (61.9%, p<0.05). In the blastocyst stage, proportion of diploid chromosome plates was significantly high in DMAP-treated group (64.2%, p<0.05), and proportion of abnormal chromosome plates was higher in CHXM-treated group (36.6%, p<0.05) than DMAP-treated group (28.3%,). Proportion of embryos with abnormal chromosome constitutions was slightly increased by DMAP (40.0%) and CHXM (42.1%) treatment due to the increasing of mixoploid (47.4 and 52.0%). The present study shows that the DMAP treatment increase the development of porcine parthenotes. However, parthenogenetic activation by ES or combined treatment with ES and DMAP or CHXM detrimentally affects the chromosome constitutions of porcine parthenotes during early embryonic development, leads to increased abnormal ploidy in the blastocyst stage.

This study was conducted to examine the viability of Korean native striped cattle (Bos namadicus Falconer, Chikso) clone embryos after embryo transfer. Chikso somatic cell nuclear transfer (SCNT) embryos were produced by fusion of ear skin cells derived from a female Chikso with enucleated oocytes matured in vitro for 18-24 hr. After in vitro culture of SCNT embryos for 7 to 8 days, fresh or vitrified blastocysts derived from SCNT were transferred into a uterine horn of recipient cows. Fifteen of total 43 recipients were pregnant at Day 50 and 4 recipients were maintained to term. Three IVF-derived calves and 1 clone Chikso calf were born. Pregnancy rate was higher when fresh embryos were transferred to recipients compared to vitrified embryos, but development to term was not different between both groups. The clone Chikso calf died at 5 days after birth due to the fullness of amniotic fluid in rumen and the infection of umbilical cord. The result of the present study shows that clone Chikso calf can produced from the embryo transfer of SCNT embryos, however, solution of abortion problem is necessary to improve the cloning efficiency.

This study was conducted to investigate the effects of oocyte maturational age and activation condition on in vitro development of porcine parthenogenetic embryos (parthenotes). Porcine follicular oocytes were matured in vitro for 30 to 44 hr. Maturation rate was examined during in vitro maturation (IVM) every 2 hr interval. The cdc2 kinase activity was measured at 36 and 44 hr of IVM. Some oocytes were activated at 36 or 44 hr of IVM by three different conditions; 1) single electric stimulation (1.5 kV/cm for 30 sec; ES), 2) double electric stimulations (1.5 kV/cm for 30 sec, followed by 1.0 kV/cm for 50 sec after 1 hr; ES+ES) or 3) ES+ES followed by culture in 6-dimethlyaminopurine (6-DMAP) for 4 hr (ES+ES+D), and cultured for 6～7 days. Maturation rate was significantly increased as culture period was increased to 36 hr (66.9%, p<0.05), and then gradually increased to 87.1% at 44 hr of IVM. The cdc2 kinase activity was decreased (p<0.05) with culture period prolonged from 36 hr to 44 hr. Lower blastocyst formation rate (4.3%, p<0.05) were obtained by ES in 36 hr-matured oocytes compared to other treatments (16.5 and 20.5%) in the same age and the same treatment in 44 hr-matured oocytes (15.0%). High blastocyst formation rate (23.6%) was obtained by ES+ES+D in 44 hr-matured oocytes (p<0.05). These results demonstrate that porcine oocyte activation and in vitro development of parthenotes can be affected by interactions between oocyte maturational age and activation condition.

The purposes of this study were to estimate Brdu positive and apoptotic cells dis tribution in co ndyl ar art icular and proliferative layer. and hi stopathological evaluat ion during one sided mas t icatory condition. 15 of 30 adults Sprague Dawley male rats were experimental group, and 15 rats were control group. Experime nta l group were gen t ly extracted on all lower and upper molar teeth to make unilateral mastication. Experimental gl'Oup were divided into two group as extracted side and nonextracted side condyle. Diluted bromodeoxyuridi ne(Brdu) solution(5mg/Kg) was injected in peri toneum before sacrifice at inte rval 1 week, 2 week, 4 week Streptoavidi n-Biotin method for Brdu was used to evaluate cellular proliferat ive activity, and fluorescent TUNEL method was used to estimate the a poptotic activity. The resul ts of this experiment were recorded about anterior and posteri or condyle sepa rately On anterior condyle, contl'Ol group was sustained increased proliferative activity th roughout experiment. whi le cel lu lar proliferative activity of extracted and nonextracted s ide condyle showed more decreased than control grou p ‘ On posteri or portion of condyle‘ control group and nonextracted group showed dramatically decreased cellular pr。 liferat ive activity during all expel‘ imental period. On anterior portion of condyle, control grollp s howed decreased a poptotic activity with time passed. bllt expel'imental gl'oup(both ext l'acted and nonextracted) exhibi ted incrcased a poptotic activity. Es pecia lly extracted grollp showed prominent increased apoptotic activity. On posteri or portion of condyle. extracted group showed dec reased apoptotic activity wi th time progress. but co ntrol and nonextracted group exhibi ted increased apoptotic activity with time progress. Conclllsively. antel'ior portion of condyle on ex t racted s ide expressed hypoplasia by low cellllJar prol iferative activity and increased apoptotic activity. and poste ri or portion on extracted side showed condylar surface hyperplasia by continious proliferative cell ular activity and low a poptotic activity. a nd t hllS unmasticatol'y condyle had anLeroposLel'iorly shorter mo rph이 ogy and verti ca11y longer morphology than masticatory and nOl'mal functioning condyle

특정 구획으로 유입된 오염물질이 해당 구획 내부에 순간적으로 균일하게 분포한다는 구획모델에 대한 기본가정의 한계로 인해, 전통적인 단일구획모델로는 불포화대에서 오염물질의 이동현상을 적절하게 예측할 수 없다. 한편 물리적으로 동일한 불포화대를 여러 개의 구획으로 구분한 다중구획모델링 기법은 실제 불포화대에서의 오염물질 이동 지연효과를 적절하게 설명할 수 있으나, 지금까지 일반적인 해석해가 보고된 바 없으며 고려하는 구획의 개수가 증가할수록 모델링에 많은 시간이 소요되는 등의 한계가 있다. 이러한 문제점을 해결하기 위하여, 이류가 지배적인 조건 하에서 불포화대의 오염물질 이동현상에 대한 다중구획모델을 수립하고 이를 해석적인 방법으로 계산할 수 있는 일반해를 유도한 후, 다중구획모델을 단일구획모델로 근사할 수 있는 수학적 제약조건을 도출하였다. 단순화된 근사방법론의 유효성은 가상적인 조건 하에서 간단한 수치해석적 방법을 통해 검증하였다. 물리적으로 동일한 특성을 갖는 불포화대를 단일 구획으로 가정할 경우, 불포화대로부터 포화대로 유입되는 오염물질의 전이율은 상수가 아닌 시간 종속적인 명목전이율로 표현할 수 있음을 증명하였다. 또한 명목전이율은 불포화대 구획간 전이율에 민감하며 오염층으로부터의 전이율에 대한 민감도는 미미한 것으로 나타났다. 이 연구에서 개발된 단순화된 근사방법론은 많은 시간이 요구되는 다중구획 모델링을 통하지 않고 불포화대 오염물질 이동현상을 신속하고 합리적으로 예측하기 위한 목적으로 활용될 수 있을 것으로 기대된다.

원자력시설 해체부지를 재이용하는 과정에서 유발될 수 있는 방사선학적 리스크를 사전에 선별하기 위한 목적으로 단순화된 방사선량 평가모델을 개발하고, 이를 Microsoft 스프레드시트와 내장된 Visual Basic 및 마크로 기능을 활용하여 기능별로 모듈화된 평가도구를 구현하였다. 이와 함께 부지 특성자료가 불충분할 경우 신속한 사전평가를 위해 적용할 수 있는 일련의 입력변수 값 목록을 제안하였다. 동일한 조건에서 이 연구에서 개발된 평가도구를 이용해 유도한 사전 선별준위가 RESRAD Ver.6.2를 이용해 계산된 유도농도지침한계 및 독일 방사선방호령에 규정된 핵종별 부지 재이용 기준농도를 합리적으로 근사할 수 있음을 확인하였다.

The purpose of this study was to examine the interfacial reaction between diamond grits and Ni-based, Ag-based, brazing filler metal, respectively. The morphology of the interface between diamond grits and Ni-based, filler metal exhibited a very good condition after this heat treatment. Cr-carbide and Ni-rich compounds were detected by XRD analysis in the vicinity of the interface between diamond grits and Ni-based, filler metal after vacuum induction brazing. Chromium carbide is considered to play an important role in the high bonding strength achieved between diamonds grits and the brazing alloy.

Thrombin-induced platelet microbicidal proteins (tPMP) are antibacterial proteins released when platelets are stimulated by thrombin. It has been reported that tPMP has antibacterial activity against various bacterial species including causative agents of infective endocarditis. Most of the oral streptococci have resistance to the killing by tPMP and this fact may play an important role as a virulence factor in infective endocarditis. However, the susceptibility and resistance mechanism of oral streptococci for tPMP have not been revealed yet. In this study, the killing mechanism of tPMP for oral streptococci has been investigated. Streptococcus rattus BHT, a susceptible strain, and Streptococcus gordonii DL1, a resistant strain, have been used in this study. tPMP was isolated from platelet after stimulation with thrombin. Cell membrane depolarization was examined with 3,3'-dipropylthiodicarbocyanine iodide (DiSC₃), membrane potential-sensitive cyanine dye, by fluorescence spectrophotometry. The permeabilization of cell membrane by tPMP was investigated with propidium iodide (PI) by flow cytometry. tPMP susceptible S. rattus BHT showed the increase of the DiSC₃fluorescence level meaning depolarization of cell membrane and increase of the uptake of PI which means permeabilization of cell membrane. However, tPMP resistant S. gordonii DLI did not show depolarization and permeabilization. These results indicate that the increasing depolarization and permeabilization of oral streptococcal cell membrane are associated with the bactericidal activity of tPMP.

Labor required for managing and harvesting the oyster mushroom bed was evaluated. Although vinyl mulching cultivation method needs more hours for spawning, it saves more than 50% of labor for harvesting and managing of the mushroom bed. Harvesting hour of 1st-3rd flush in vinyl mulching method was 48~50% for Plerutus ostreatus and 36~41% for P. sajor-caju. Labor for bed management after harvesting in vinyl mulching method was also 38~50% for P. ostreatus compared to conventional method, and 20~35% for P. sajor-caju. Vinyl mulching is believed to be a very efficient method for saving labor in oyster mushroom cultivation.