• Journal of Embryo Transfer
• /
• v.19 no.2
• /
• pp.155-163
• /
• 2004

The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.45 no.4
• /
• pp.484-491
• /
• 2016

The present study investigated the effect of Sinetrol-XPur (polyphenolic Citrus spp. and Paullinia cupana Kunth dry extract) and defined the action mode for cyclic adenosine monophosphate (cAMP)-dependent uncoupling protein (UCP)-2 activation. Leptin-deficient obese mice were treated with two different doses, 100 mg/kg body weight (BW) and 300 mg/kg BW of each AIN93G supplement, for 7 weeks. Treatment of obese mice with both low and high doses of Sinetrol-XPur significantly reduced body weight gain compared to control obese mice. White adipose tissue weight of mice was reduced by 30.96% in high dose-supplemented groups. Serum total cholesterol and triglyceride were reduced by a high dose of Sinetrol-XPur by 20.02% and 30.96%, respectively. Serum level of high density lipoprotein (HDL) was significantly increased by treatment with both doses, as the ratio of HDL to low density lipoprotein increased by 138.78% and 171.49%, respectively. Regarding expression of biochemical factors related to lipid metabolism, fatty acid synthase significantly decreased and UCP-2 increased upon treatment with a high dose of Sinetrol-XPur, but there was no significant difference in lipoprotein lipase and hormone-sensitive lipase. To define cellular mechanism, intracellular cAMP levels in 3T3-L1 adipocytes significantly increased in a dose-dependent manner over the range of $50{\sim}250{\mu}m/mL$. The phosphodiesterase (PDE) inhibitor 3-isobutyl-1-methylxanthine clearly blocked cAMP, suggesting that Sinetrol-XPur promotes lipolysis of adipocytes through inhibition of cAMP-dependent PDE, resulting in induction of cAMP response element binding protein and UCP-2. These results suggest that Sinetrol-XPur supplementation is a viable option for reducing body weight and fat by improving serum lipid profiles and genetic expression of lipid metabolic factors, especially activation of cAMP-dependent UCP-2.

• Korean Journal of Veterinary Research
• /
• v.32 no.2
• /
• pp.157-164
• /
• 1992

Changes in the both inward current and conductance of membrane by the fertilization were observed using the one microelectrode voltage clamp(or switch clamp) technique. Unfertilized eggs and both 1- and 2-cell stage eggs after fertilization were donated from the superovulated mouse (ICR, more than 6 weeks old) treated with PMSG(pregnant mare serum gonadotropin, Sigma) and HCG(human chorionic gonadotropin, Sigma) and naturally mated ones, respectively in this experiment. Membrane potential was held at -90mV and the voltage step was applied from -80mV to 50mV with interval of 10mV or 20mV for 300ms. since both of amplitudes and time courses in the membrane currents were various according to the states of cells and clamping condition, results were presented by their $averages{\pm}SEM$(standard mean error)and ratios or percentages. Inward currents began to appear in response to the step depolarization from -60mV and reached its maximum at -50mV. However, since the potential was not clamped evenly during the voltage step, current-voltage(I-V) relationship might be positively shifted 10 or 20mV. From the steady-state currents plotted in the I-V curve, outward rectification was markedly observed. Peak inward currents$(i_{in})$ at -50mV were $-0.62{\pm}0.23nA$(n=4),$-0.52{\pm}0.25nA$(n=5) and $-0.37{\pm}0.25nA$(n=6), in the 1-cell stage, 2-cell stage fertilized eggs and in the unfertilized eggs, respectively. Pure inward current (difference between steady-state and peak, $i_{in. pure}$) were $-1.01{\pm}0.23nA$, $-0.69{\pm}0.43nA$ and $-0.68{\pm}0.29nA$, respectively in the 1-cell stage fertilized eggs, unfertilized eggs and 2-cell stage fertilized eggs. These results suggested that the outward current in fertilized eggs of 2-cell stage was more increased than those in the unfertilized eggs. Pure inward currents in the all stages of eggs showed a similar fashion in the I-V relationship from -50mV to 50mV and reversal potential at 50mV. Time constant of inactivation$({\tau})$ in the inward current was decreased as the membrane potential was depolarized in the unfertilized and 2-cell stage eggs but in the 1-cell stage eggs t was not likely to be affected significantly. Slope conductances were 14.2nS, 8.9n5 and 7.7nS in the 1-cell, 2-cell stage fertilized eggs and the unfertilized eggs, respectively. Membranes between two cells within a zona pellucida seem to be electrical-connected in the 2-cell stage eggs from the observation made in the analysis for the electronic spread and decay to the current stimuli. Both of inward current and membrane conductance were increased after fertilization in the mouse eggs. Inward current seems to be carried by the same ion or through the same channels up to the 2-cell stage and ion that carried inward current was thought to play important function after fertilization in the mouse eggs.

• Tuberculosis and Respiratory Diseases
• /
• v.47 no.1
• /
• pp.50-56
• /
• 1999

Background : Pleural effusion is a common clinical problem and many clinical and laboratory evaluations, such as tumor marks, have been studied to discriminate malignant pleural fluid from benign pleural fluid. However their usefulness in the diagnosis of pleural effusion is still not established fully. We studied the diagnostic value of cyfra 21-1 in diagnosis of malignant pleural effusion. Methods: Pleural fluid was obtained from 45 patients with malignant diseases(32 lung cancer patients, 13 metastatic malignant diseases) and 47 patients with benign diseases. The level of cyfra 21-1 in the pleural fluid and serum were determined using a CYFRA 21-1 enzyme immunoassay kit(Cis-Bio International Co.). The t-test was used for comparison between two diseases groups and receiver operating characteristic(ROC) curves were constructed by calculating the sensitivities and specificities of the cyfra 21-1 at several points to determine the diagnostic accuracy of the cyfra 21-1. Results: In patients with primary lung cancer, the level of cyfra 21-1 in the pleural fluid was significantly higher than those of patients with benign diseases and had positive correlations between the level of cyfra 21-1 in the pleural fluid and serum levels. In the ROC curve analysis of the pleural fluid, the curve for primary lung cancer group was located closer to the left upper comer and the cut off value, sensitivity and specificity of the cyfra 21-1 of the primary lung cancer group was determined as 22.25ng/ml, 81.8% and 78.7% respectively. Conclusions: Our data indicates that the measurement of cyfra 21-1 level in pleural effusion has useful diagnostic value to discriminate malignant pleural effusion in primary lung cancer from benign pleural effusion.

• Clinical and Experimental Pediatrics
• /
• v.46 no.5
• /
• pp.467-473
• /
• 2003

Purpose : The Epstein-Barr virus(EBV), gamma herpesvirus, is an important pathogen that is widespread around the world. The EBV causes various diseases depending on the geographic location, and on the immunity or the premorbid condition of the person exposed to EBV. To evaluate EBV typing may be the most important step to figure out the pathogenesis of EBV associated diseases, and we need to re-evaluate the pathologic role of human leukocyte antigen(HLA) in developing Epstein- Barr virus associated acute infectious mononucleosis by using newly developed methods. Methods : This study included 24 children(age range : 6 to 13 years), serologically confirmed with acute infectious mononucleosis. The control group for the HLA type consisted of 200 age-matched healthy children. To classify HLA I, modified ARMs-PCR was used, while modified PCR-SSOP was utilized in typing of HLA II. Also, we performed EBV typing in study patients by using a one-step PCR. Results : The results of HLA types : In HLA class I, HLA-A24 was positive in 69 of 200 healthy children and positive in 14 of 24 patients in the study group(relative risk : 3.5724, chi-square; 5.26, P<0.05). In HLA class II, HLA-DRB1*07 was detected in 18 of 200 healthy children, and eight of 24 patients in the study group(relative risk; 506173, chi-square; 9.73, P<0.01). The results of EBV types : In the research group, 20(83.8%) of 24 patients were shedding type A virus, while 4(16.7%) were type B. Conclusion : We conclude that development of infectious mononucleosis may be associated with HLA types, and these results suggest that acute infectious mononucleosis could have hereditary traits. And we confirm that type A EBV is highly prevalent in patients with acute infectious mononucleosis in Korea. Also, our results suggest that further large scale studies, including adult groups, regarding the association between pathogenesis of EBV with HLA-DP or HLA-DQ will be warranted.

• Radiation Oncology Journal
• /
• v.13 no.2
• /
• pp.149-156
• /
• 1995

Purpose : Since February 1991 a Prospective study for non-small cell lung cancer patients who underwent radical resection and had a risk factor of positive resection margin or regional lymph node metastasis has been conducted to evaluate the effect of MVP chemotherapy and radiotherapy on the pattern of failure, disease free and overall survival. and tolerance of combined treatment. Materials and Methods: Twenty nine patients were registered to this study until Sep. 1993; of these 26 received planned therapy Within 3 weeks after radical resection, two cycles of MVP(Mitomycin C $6mg/m^2,$ Vinblastin $6mg/m^2,$ Cisplatin $60mg/m^2$) chemotherapy was given with 4 weeks intervals. Radiotherapy (5040cGy tumor bed dose and 900cGy boost to high risk area) was started 3 to 4 weeks after chemotherapy. Results: One and two year overall survival rates were $76.5\%\;and\;58.6\%$ respectively. Locoregional failure developed in 6 patients$(23.1\%)$ and distant failure in 9 patients$(34.6\%)$ Number of involved lymph nodes, resection margin positivity showed some correlation with failure pattern but T-stage and N-stage showed no statistical significance. The group of patients who received chemotherapy within 2 weeks postoperatively and radiotherapy within 70 days showed lower incidence of distant metastasis. Postoperative combined therapy were well tolerated without definite increase of complication rate, and compliance rate in this study was $90\%$. Conclusion: 1) MVP chemotherapy showed no effect on locoregional recurrence, but appeared to decrease the distant metastasis rate and 2) combined treatments were well tolerated in all patients. 3) The group of patients who received chemotherapy within 2 weeks postoperatively and radiotherapy within 70days showed lower incidence of distant metastasis. 4) Addition of chemotherapy to radiotherapy failed to increase the overall or disease free survival.

• Clinical and Experimental Reproductive Medicine
• /
• v.33 no.4
• /
• pp.237-243
• /
• 2006

Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.

• Journal of Yeungnam Medical Science
• /
• v.16 no.1
• /
• pp.94-100
• /
• 1999

The aim of this retrospective study was to determine whether a nephron sparing surgery might be feasible in patients with a small solid renal tumor. Materials and methods: Between 1988 and 1999, 21 patients with a radiologically detectable small solid renal tumor underwent enucleoresection, wedge resection and polar segmental nephrectomy. The mean age of the 11 men and 10 women in this study was 43 years (range 14 to 68). According to the preoperative radiological diagnosis, 15 among the 21 patients were considered to have renal cell carcinoma, 4 were considered to have angiomyolipoma, and in the remaining 2 patients, radiological differentiation of renal tumors was difficult. Among 15 patients considered to have renal cell carcinoma, 14 were found to have renal cell carcinoma and the remaining one patient was diagnosed as having oncocytoma on pathologic examination. Radiological determination of angiomyolipoma in four patients was confirmed to be correct on pathological examination. The 2 patients whose radiological diagnose was difficult were found to have cavernous hemangioma and angiomyolipoma. One patient with renal cell carcinoma developed arteriocaliceal fistula, the only immediate complication in this series and underwent nephrectomy on postoperative 10th day. The mean follow up duration for the 14 patients with renal cell carcinoma was 18.6 months (range:1-103). There was no other tumor involvement in the resection margins following the nephron sparing surgery. These results suggest that nephron sparing surgery provides an effective treatment for patients with a single, small, unilateral, localized renal tumor. Longer follow-up is suggested for more definite verification of the role of nephron sparing surgery.

• Food Science and Preservation
• /
• v.23 no.3
• /
• pp.402-412
• /
• 2016

Generation of biogenic amines (BAs) and physiological activities of Makgeolli, Korean rice wine, were investigated during fermentation under different temperatures (20, 25, and $30^{\circ}C$) and time (3, 5, and 7 days). The pH was 3.96~4.36 during fermentation and the acidity increased proportionally with temperature and time. Alcohol and total phenolic contents peaked on day 5 at 25 by 9.0~9.8% and $3.01{\pm}0.07mg/g$, respectively. The organic acid mostly produced was lactic acid, which increased gradually with temperature and time. Formation of biogenic amines comprised of tryptamine and putrescine was observed during fermentation: 6.91, 11.70, and 15.63 mg/mL of tyramine on day 7 at 20, 25, and $30^{\circ}C$, respectively. Antioxidant activities from DPPH and FRAP assay were high in the order of $25^{\circ}C$>$20^{\circ}C$>$30^{\circ}C$ while that from ABTS assay was high regardless of temperature. Processed at 1 mg/mL and $30^{\circ}C$, the inhibitory effect on ${\alpha}$-amylase was $67.99{\pm}0.11$, $73.64{\pm}0.43$, and $75.51{\pm}0.26%$ on days 3, 5, and 7, respectively, which increased proportionally with temperature and time. Inhibitory activity on lipid accumulation in 3T3-L1 adipocytes was the highest in Makgeolli fermented on day 5 at $25^{\circ}C$. This study shows that fermentation at $25^{\circ}C$ for 5 days yields Makgeolli of the best quality with high bioactivities but no biogenic amines.

• IMMUNE NETWORK
• /
• v.3 no.2
• /
• pp.150-155
• /
• 2003

Background: We investigated the effect of donor marrow T cell depletion, administration of FK506, cyclosporin A (CSA), and 3-deazaadenosine (DZA) on graft versus host disease (GVHD) after allogeneic murine hematopoietic stem cell transplantation (HSCT). Methods: We used 4 to 6 week old Balb/c ($H-2^d$, recipient), and C3H/He ($H-2^k$, donor) mice. Total body irradiated recipients received $1{\times}10^7$ bone marrow cells (BM) and $0.5{\times}10^7$ splenocytes of donor under FK506 (36 mg/kg/day), CSA (5 mg/kg/day, 20 mg/kg/day), and DZA (45 mg/kg/day), which were injected intraperitoneally from day 1 to day 14 daily and then three times a week for another 2 weeks. To prevent the GVHD, irradiated Balb/c mice were transplanted with $1{\times}10^7$ rotor-off (R/O) cells of donor BM. The severity of GVHD was assessed daily by clinical scoring method. Results: All experimental groups were well grafted after HSCT. Mice in experimental group showed higher GVHD score and more rapid progression of GVHD than the mice with R/O cells (R/O group) (p<0.01). There were relatively low GVHD scores and slow progressions in FK506 and low dose CSAgroups than high dose CSA group (p<0.01). The survival was better in FK506 group than low dose CSA group. All mice treated with CSA died within 12 days after HSCT. The GVHD score in DZA group was low and slow in comparison with control group (p<0.05), but severity and progression were similar with low dose CSA group (p=0.11). All mice without immunosuppressive treatment died within 8 days, but all survived in R/O group (p<0.01). Survival in low dose CSA group was longer than in control group (p<0.05), but in high dose CSA group, survival was similar to control group. The survival benefit in DZA group was similar with low dose CSA group. FK506 group has the best survival benefit than other groups (p<0.01), comparable with R/O group (p=0.18), although probability of survival was 60%. Conclusion: We developed lethal GVHD model after allogeneic murine HSCT. In this model, immunosuppressive agents showed survival benefits in prevention of GVHD. DZA showed similar survival benefits to low dose CSA. We propose that DZA can be used as a new immunosuppressive agent to prevent GVHD after allogeneic HSCT.