• Journal of Embryo Transfer
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• v.22 no.1
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• pp.53-61
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• 2007

This study was investigated factors affecting the pregnancy rates after transfer of pronuclear microinjected embryos for the production of transgenic Korean black goats. Embryo transfer was carried out in 343 recipient Korean black goats from September 1999 to June 2000. Estrus was induced by the insertion of intravaginal progesterone devices $CIDR^(R)$ for 2 weeks. A single injection of 400 IU equine chorionic gonadotropin was administered at 48h before $CIDR^(R)$ removal to increase the proportion of does cycling and ovulation rate. Good quality embryos were prepared by microinjection of DNA into the pronuclei of fertilized goat oocyte and cultured in vitro. Pronuclear microinjected $1{\sim}8$ cell stage embryos were surgically transferred into the oviducts of the recipient at day 4 or 5 following $CIDR^(R)$ removal, and morula to blastocyst stage embryos were surgically transferred into uterus at day 9. Pregnancy was diagnosed by transrectal ultrasound scanning at $20{\sim}30d$ and 8 weeks following embryo transfer. The pregnancy rate was affected by several factors, such as estrus induction, the number of previous transfer, transfer site, stage of CL (corpus luteum), the number of recipient CL, stage of embryos and the number of transferred embryo. The pregnancy rate was significantly higher in recipients that came into estrus naturally than recipients that induced to come into estrus with $CIDR^(R)$(59.1% vs. 36.8%; P<0.05). The pregnancy rate was higher when the embryos were transferred into the left oviduct than transferred into the right oviduct (42.9% vs. 35.3%; P<0.05). The pregnancy rate of recipients with $CH_1$ (early) stage corpus hemorrhagicum in ovary was hi틴or than recipient with $CH_3$ (late) stage hemorrhagicum (47.5% vs. 17.9%; P<0.01). Higher pregnancy rates were obtained by transfer of 1-cell stage embryos into oviduct while late blastocysts (51.6% vs. 66.7%; P<0.01) into uterus. The pregnancy rates when 3 embryos were transferred to recipients were significantly higher than when 2 embryos we.e transferred (47.6% vs. 27.0%; P<0.05). Although there were no significant difference among the group, adhesion of reproductive organs, uterine size, ovulation rate of recipients, presence of large follicle and difficulty of transfer affected pregnancy rate of recipient. Higher pregnancy rates were obtained in the recipients with $8{\sim}15m$ diameter uterine horn as compared to the recipients with <5m diameter or >20mm diameter uterine hem (38.9%, 20% vs. 18.2%), in the recipients with large follicle in the ovulated ovary ipsilaterally (53.6% vs. 37.1%) and in the transfer which was carried out easily (39.2% vs. 27.8%, 0%). In conclusion, the high rate of pregnancy was achieved following transfer of pronuclear microinjected embryos when three or four 1-cell stage embryos were transferred into oviduct with $CH_1$ stage corpus hemorrhagicum in the ovary of recipient which came into estrus naturally.

• Tuberculosis and Respiratory Diseases
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• v.63 no.3
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• pp.251-260
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• 2007

Purpose: An imbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis. Capase 3, survivin and p53 have been identified as important members of the apoptotic related proteins. This study evaluated the proliferating cell nuclear antigen(PCNA), apoptosis, apoptotic related protein such as capase 3, survivin and p53 using urethane-induced mouse lung carcinogenesis, which provides reproducible steps from hyperplasia to adenocarcinoma. Methods: Urethane was administered to the ICR mice through an intra-peritoneal injection, The mice were sacrificed at 5, 15, and 25 weeks after urethane intervention. The sequential morphological changes and immunohistochemical expression of PCNA, apoptosis, capase 3, survivin, and p53 were examined during mouse lung carcinogenesis. Results: During carcinogenesis, the sequential histological changes were observed from hyperplasia of type II pneumocytes, to anadenoma, and ultimately to an overt adenocarcinoma. The PCNA Labeling index (LI) was 9.6% in hyperplasia, 23.2% in adenoma, and 55.7% in adenocarcinoma, respectively. The apoptotic LI was 0.24% in hyperplasia, 1.25% in adenoma, and 5.27% in adenocarcinoma. A good correlation was observed between the PCNA LI and apoptotic LI. The expression of caspase 3 was remarkable- i.e., 46.7% in adenocarcinoma, in contrast to 15% in hyperplasia and 16% in adenoma. Survivin was detected weakly in the alveolar hyperplasia and showed an increasing expressional pattern in adenoma and adenocarcinoma. p53 expression was detected only in the adenocarcinoma lesions with an expression rate of 13.3%. The level of caspase 3 expression correlated with the increase in the apoptotic index. The positive expression of caspase 3 was associated with an increased apoptotic index. Conclusions: These results suggest that the PCNA LI and apoptotic LI might be useful markers for evaluating the risk of a malignant transformation. In addition, caspase, survivin and p53 might play a role in the early and late steges of urethane-induced mouse lung carcinogenesis.

• Korean Journal of Agricultural Science
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• v.12 no.2
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• pp.242-252
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• 1985

In order to get information on the ecology of rice false smut, germination ability and pathogenicity of sclerotia and chlamydospores of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. The degree of outbreak of rice false smut was higher in the upland rice in comparison with the paddy field rice in respect to the number of affected grains per ear, the size and weight of smut balls formed on affected grains as well as the ratio of sclerotial formation produced on smut balls. 2. Germination percentage and days required for germination of overwintered sclerotia placed on the soil surface in July were 81% and 19 days, respectively, while those of overwintered sclerotia treated in May were 60-70% and 41 days. Sclerotia placed on the soil surface or under 1 cm depth of the soil surface and incubated at $25-30^{\circ}C$ were germinated well, whereas those placed under 3 cm or 5 cm depth of the soil surface were not germinated at all. Germinability and stroma productivity of sclerotia were reduced when the sclerotia were cutted into small pieces. 3. The average number of stroma formed on a sclerotium was six and that of perithecia formed in a stroma was about 50 to 140. 4. Percentage of germination of chlamydospores on the yellow balls was very high and was decreased as the color of the balls being darken with maturation. 5. Panicle of rice plants were successfully infected by injection inoculation with suspention of ascospores and chlamydospores of the pathogen to the sheaths at the booting stages, while seeding infection by spraying with suspensions of chlamydospores was unsuccessful. 6. More number of infected grains was distributed on basal parts of an affected ear than that of infected ones distributed upper parts of the ear, when the affected ear was divided into five parts from its basal portion to the apical of the ear. 7. The occurrence of the disease was more severe in the late maturing varieties of rice in comparison with the early maturing varieties. 8. When the level of nitrogen applied was increased, the incidence of disease increased, and the infection percentage of the disease was increased as the transplanting date was delayed. 9. The weight of panicles and 1000 kernels and the ratio of ripenness were reduced, and the contamination degree of grains with chlamydospores were increased as the number of smut balls per panicle were increased.

• Tuberculosis and Respiratory Diseases
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• v.43 no.4
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• pp.579-589
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• 1996

Background : Activation of neutrophil is critical for the clearance of microorganisms and toxic host mediators during sepsis. Unfortunately the activated neutrophil and its toxic byproducts can produce tissue injury and organ dysfunction. The leucocyte CD11/18 adhesion complex regulates neutrophil-endothelial cell adhesion, the first step in neutrophil migration to sites of injection and inflammation. To investigate the potential of neutrophil inhibition as a treatment strategy for sepsis, we evaluated the effects of monoclonal antibody against CD11b (MAb 1B6) in rats intrabronchial challenged with Escherichia coli. Methods : Animals were randomly assigned to receive monoclonal antibody against CD11b (1 mg/kg, sc) and bovine serum albumin(BSA, 1 mg/kg, sc) 6 hr before, at 0 and 6 hr after intrabronchial challenge of $20x10^9$ CFU/kg E. coli 0111. Animals were randomized to treat either 24, 60 or 90% oxygen after bacterial challenge and begining 4 hr after inoculation, all animals were received 100 mg/kg ceftriaxone qd for 3 days. Peripheral and alveolar neutrophil(by bronchoalveolar lavage) counts and lung injury parameters such as alveolar-arte rial $PO_2$ difference, wet to dry lung weight ratio and protein concentration of alveolar fluid were measured in survived rats at 12 hr and 96 hr. Results : Monoclonal antibody against CD11b decreased circulating and alveolar neutrophil especially more in 12 hr than in 96 hr The lung injury parameters of antibody-treated animals were not different from those of BSA-treated animals. but It was meaningless due to small number of survived animals. The early(6 hr) mortality rate was significantly increased in antibody-treated group(51%) compared to BSA-treated group(31%) (P=0.02) but late(from 12 hr to 72 hr) mortality rate was not different in antibody-treated group(44%) from BSA-treated group(36%) (P =0.089). Conclusion : Leucocyte CD11b/18 adhesion molecule is known to regulate neutrophil migration to the site of infection and inflammation. The monoclonal antibody against CD11b decreased alveolar neutrophil in rats with pulmonary sepsis and increased early mortality rate. Therefore, we can speculate that monoclonal antibody against CD11b blocks of alveolar recruitment of neutrophils, impairs host defense mechanism and increases early mortality rate of pulmonary sepsis in rat.

• Journal of Korean Neurosurgical Society
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• v.29 no.11
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• pp.1421-1428
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• 2000

Objectives : The aim of this study is to evaluate the sequential metabolic changes in experimental hydrocephalus and the clinical applicability to the diagnosis and prognosis of hydrocephalus using proton MR spectroscopy. Methods : Hydrocephalus was experimentally induced in 30 cats(2-3kg body weight) by injecting 1ml of sterile kaolin suspension(250mg/ml) into the cisterna magna. Proton MRS was performed with a 1.5 T MRI/MRS unit (Vision Plus, Siemens) at pre-treatment and at 1, 3, 7, 14, 21, and 28 days after the kaolin injection. PRESS(TR/TE＝1500/270msec) technique was employed. The major metabolites which include N-acetyl aspartate (NAA), creatine(Cr), choline(Cho), and lactate(Lac) were quantitatively analyzed and the relative concentrations ratios were evaluated. Multislice $T_2$-weighted images were also obtained using fast spin echo sequence(TR/TE＝ 2500/96msec) to monitor the morphologic changes along with progression of hydrocephalus. Results : Hydrocephalus was successfully induced in all 30 cats. Twenty five cats died within 3 days and one at the end of the second week. In all animals, the NAA/Cr ratios initially decreased during the acute stage. In 4 surviving cats, the NAA/Cr ratios initially decreased during the acute stage(<14 days) and then gradually increased to the prekaolin level as follows : pre-kaolin($1.49{\pm}0.04$), day 1($1.11{\pm}0.07$), day 7($1.17{\pm}0.04$), day 14($1.40{\pm}0.03$), day 21 ($1.46{\pm}0.06$), day 28($1.43{\pm}0.03$). These levels were relatively well correlated with the symptomatologic improvement. Lactate peak, which reflects the evidence of ischemia, did not appear throughout the entire period except in one case which expired at the end of the second week. Conclusions : The NAA/Cr ratio of the sequential proton MRS in kaolin-induced hydrocephalic cats reflects a metabolic aspect of the hydrocephalus at each stage. A decreased NAA level at the early stage is from both neuronal and axonal damage which may provide diagnostic information in the acute stage of hydrocephalus. In addition, the initial fall of NAA/Cr ratio and recovery in the late stage, when no lactate peak emerges, may suggest that the main insult of the parenchyma is not to the neuron itself but to the axon, which may be related to a good prognosis. However, emergence of the lactate peak and unrecoverable NAA/Cr at the end of the acute phase may be a poor prognostic factor. In the chronic stage, recovery of NAA/Cr ratio may provide a diagnostic clue for the differentiation between hydrocephalus and cortical atrophy.

• The Korean Journal of Nuclear Medicine
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• v.33 no.3
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• pp.247-261
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• 1999

Purpose: The aim of this study was to evaluate the normal values of regional cerebral blood flow (rCBF) and cerebrovascular reserve (CVR) in normal children to aged volunteers using Tc-99m HMPAO, Materials and Methods: Thirty four right-handed normal volunteers (20 males, 14 females, mean age $40.3{\pm}24.9$ years, range 4 to 82 years) were underwent rost/acetazolamide (ACZ) brain SPECT using Tc-99m HMPAO and the sequential injection and subtraction method. rCBF was estimated on the basis of a semiquantitative approach by means of right/left ratio, region/cerebellum and region to whole brain ratios in (rental, parietal, temporal, and occipital lobes, basal ganglia, thalami, and cerebellum. CVR was measured by means of % perfusion increase calculated as % mean count change compared to rest rCBF in each regions. Results: Mean values of right to left ratios range from 1.004 to 1.018, rCBF was highest in cerebellum and lowest in basal ganglia and thalami. Frontal and temporal rCBF decreased while occipital and thalamic rCBF increased according to age. No sexual difference of rCBF was noted. Mean CVR was $29.9{\pm}12.9%$. Mean CVR significantly increased to late teens, and declined thereafter. After 6th decade, CVR in both frontal lobes, left parietal lobe and right basal ganglia decreased significantly with advancing age. There was no sexual difference of CVR. Conclusion: Quantitative assessment of CVR was possible by ACZ Tc-99m HMPAO brain SPECT. It revealed that rCBF and CVR changed according to age in normal Korean volunteers. There was no sexual difference.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.548-555
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• 2018

Objective: This experiment was conducted to investigate whether asparagine (Asn) could improve liver energy status in weaning pigs when challenged with lipopolysaccharide. Methods: Forty-eight weaned pigs ($Duroc{\times}Large\;White{\times}Landrace$, $8.12{\pm}0.56kg$) were assigned to four treatments: i) CTRL, piglets received a control diet and injected with sterile 0.9% NaCl solution; ii) lipopolysaccharide challenged control (LPSCC), piglets received the same control diet and injected with Escherichia coli LPS; iii) lipopolysaccharide (LPS)+0.5% Asn, piglets received a 0.5% Asn diet and injected with LPS; and iv) LPS+1.0% Asn, piglets received a 1.0% Asn diet and injected with LPS. All piglets were fed the experimental diets for 19 d. On d 20, the pigs were injected intraperitoneally with Escherichia coli LPS at $100{\mu}g/kg$ body weights or the same volume of 0.9% NaCl solution based on the assigned treatments. Then the pigs were slaughtered at 4 h and 24 h after LPS or saline injection, and the liver samples were collected. Results: At 24 h after LPS challenge, dietary supplementation with 0.5% Asn increased ATP concentration (quadratic, p<0.05), and had a tendency to increase adenylate energy charges and reduce AMP/ATP ratio (quadratic, p<0.1) in liver. In addition, Asn increased the liver mRNA expression of pyruvate kinase, pyruvate dehydrogenase, citrate synthase, and isocitrate dehydrogenase ${\beta}$ (linear, p<0.05; quadratic, p<0.05), and had a tendency to increase the mRNA expression of hexokinase 2 (linear, p<0.1). Moreover, Asn increased liver phosphorylated AMP-activated protein kinase (pAMPK)/total AMP-activated protein kinase (tAMPK) ratio (linear, p<0.05; quadratic, p<0.05). However, at 4 h after LPS challenge, Asn supplementation had no effect on these parameters. Conclusion: The present study indicated that Asn could improve the energy metabolism in injured liver at the late stage of LPS challenge.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.31-37
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• 2000

This study was designed to identify the ultrastructural changes of mouse endometrium during peri-implantation period and obtain the fundamental information for the establishment of 3-dimensional culture system of mouse endometrial cells in vitro. The used female ICR mice ($6{\sim}8$ wks) were conducted on pregnant. The biopsies were obtained from whole uterus at cycle day 1 (D1) and day 5 (D5) after hCG injection and mating. The biopsies materials were fixed 2.5% glutaraldehyde and 1% osmium tetroxide. Subsequently, for observation using light and transmission electron microscopy (LM and TEM), they were dehydrated and embedded in Epon and the embedded biopsies were sectioned and stained. For scanning electron microscopy (SEM), the fixed specimens were dehydrated, dried and coated with gold. 1) For LM, the biopsied materials at D5 (late secretory phase) were appeared the extended stromal layer by increased connective tissues and the fully developed endometrial glands and vessels compared with D1 (early secretory phase). 2) For TEM, the mouse endometrium was consisted of 3-layers, a simple polarized columnar epithelial cells, basement membrane and stromal cells. At D5, the distribution of microvilli, endoplasmic reticulum, Golgi body, lipid and glycogen deposits, secretory granules and surface area of basement membrane were increased. 3) For SEM, the degree of folding and microvilli of surface of mouse epithelial cells was became more and more according to the process of secretory phase, and at D5, implantation time of mouse, the appearance of pinopodes as a specific marker of uterine receptivity was found. The uterine pinopodes of mouse were found in narrow sites at the luminal surface, irregularity and appeared the different stages in the same sample. Therefore, these results indicated that the mouse endometrium was experienced dramatic morphological changes during peri-implantation period.

• Journal of The Korean Ophthalmological Society
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• v.58 no.6
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• pp.706-711
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• 2017

Purpose: To analyze blinking patterns when watching an ultra-high definition (UHD) television and to compare the results between normal eyes and dry eyes. Methods: A total of 59 participants aged from 13 to 69 years were instructed to watch a colorful and dynamic video on a UHD television for 10 minutes. Before and after watching the UHD television, we measured the best corrected visual acuities, autorefraction, tear-break-up-time, degree of corneal erosion and conjunctival hyperemia via slit lamp biomicroscopy. In addition, questionnaires for the evaluation of eye fatigue and symptoms of a dry eye were completed. The definition of dry eye syndrome was that the tear-break-up-time of one of the eyes was less than 5 seconds, conjunctival injection, or marked corneal erosion. The number of blinks and the duration of blinking were both measured and analyzed at the early and late phases of video-watching. Results: After watching the UHD television in the normal eye group, the tear-break-up-time was significantly decreased (p < 0.001) and the degree of corneal erosion was significantly increased (p = 0.023). However, the subjective symptoms of participants were not aggravated (p = 0.080). There were no significant differences in blinking patterns in the dry eye group. On the other hand, in the normal eye group, the mean blinking time was significantly increased (p = 0.030). Conclusions: Watching an UHD television changes the tear-break-up-time, degree of corneal erosion, and blinking pattern in normal eyes, which may increase the risk of dry eye syndrome.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.85-92
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• 1999

This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.