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PREGNANCY RATE AND SURVIVAL IN CULTURE OF IN VITRO FERTILIZED BOVINE EMBRYOS FROZEN IN VARIOUS CRYYOPROTECTANTS AND THAWED USING A ONE-STEP SYSTEM

  • Suzuki, T.;Takagi, M.;Yamamoto, M.;Boediono, A.;Saha, S.;Sakakibara, H.;Oe, M.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 1997.05a
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    • pp.27-34
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    • 1997
  • Bovine oocytes surrounded with compact cumulus cells were cultured for 20 to 22 hours($38^{\circ}C$, 5% $CO_2$) in modified TCM-199 medium supplemented with 5% superovulated cow serum(SCS) and inseminated by in vitro capacitated spermatozoa. Day 7 to 8 embryos were equilibrated for 10 minutes in 1.3M methyl cellosolve(MC) <1.1M diethylene glycol(DEG), 1.8M ethylene glycol(EG), 1.6M propylene glycol(PG) and 1.1 M 1,3-butylene glycol(BG) solutions. They were then loaded into 0.25ml straws, placed into an alcohol bath freezer at $0^{\circ}C$, cooled from $0^{\circ}C$ to $-6^{\circ}C$ at $-1^{\circ}C$/minute, seeded, held for 10 minutes, and stored in liquid nitrogen. After thawing in $30^{\circ}C$ water, the embryos wee rehydrated in TCM-199 medium and then cultured for 48 hours in TCM-199 plus 5% SCS. Embryos were considered viable if they progressed to later developmental stages with a good morphology. Some of the embryos frozen in each cryoprotectant were thawed and transferred non-surgically without removing the cryoprotectant. Hatched embryos survived freezing and one-step dilution as follows : EG(50.0%), MC(53.6%), DEG(56.9%), PG(58.0%) and BG(11.5%). The survival rate of embryos cooled at -0.3^{\circ}C$ vs. $-0.5^{\circ}C$/minute was not significantly different(P<0.05), however, blastocysts hatched most often (P<0.01) in vitro when cooled at a rate of $0.3^{\circ}C$/minute(64.6%), 31/48) than at $-0.5^{\circ}C$/minute(22.6%, 12/53). Pregnancy rates resulting from embryos frozen in the different cryoprotectants were as follows : MC(48%, 10/21); DEG(30%, 3/10); EG(74%, 20/27); and PG(40%, 4/10). These results indicate that MC, DEG, EG and PG have utility as cryoprotectants for the freezing and thawing of IVF Bovine embryos.

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Relative Importance of Bottom- up vs. Top-down Controls on Size-structured Phytoplankton Dynamics in a Freshwater Ecosystem: I. Temporal and Spatial Variations of Size Structure (담수성 식물플랑크톤의 크기별 동태에 대한 상향식, 하향식 조절간의 상대적 중요도 조사: I. 크기구조의 시 ${\cdot}$ 공간적 변동)

  • Kim, Jong-Min;Sin, Yong-Sik
    • Korean Journal of Ecology and Environment
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    • v.36 no.4 s.105
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    • pp.403-412
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    • 2003
  • Temporal and spatial variations of size-structured phytoplankton (chlorophyll a) were investigated over an annual cycle (February-October, 2003) to elucidate phytoplankton dynamics in the Juam Reservoir, Chonnam. Physical properties were also measured to investigate the relationship between the properties and temporal and spatial variations of size structured phytoplankton using simple linear regression. Phytoplankton (chlorophyll a) were grouped into three size classes: micro-size(> 20 ${\mu}m$), nano-size (3-20 ${\mu}m$) and pico-size (< 3 ${\mu}m$) in this study. Physical properties included water temperature, light attenuation coefficients, PAR (photosynthetically active radiation) and turbidity. Maximum chlorophyll a was observed in April, 2003 in the lower region whereas a peak of chlorophyll a developed in October, 2003 in the upper region. Large cell-sized phytoplankton (micro-size class)were dominant in the events of the chlorophyll a peaks. Potential mechanisms in the physical properties affecting the size-structured phytoplankton dynamics in the Juam Reservoir were discussed.

Denitrification Potential and Denitrifier Abundance in Downstream of Dams in Temperate Streams (온대지역 댐 하류의 탈질화 능력과 탈질화균 분포)

  • Vo, Nguyen Xuan Que;Lee, Seung-Hoon;Doan, Tuan Van;Jung, Sokhee P.;Kang, Hojeong
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.137-151
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    • 2014
  • Various studies have been conducted to investigate effects of dams on river ecosystems, but less information is available regarding damming impacts on downstream denitrification. We measured denitrification enzyme activity (potential denitrification rate) and denitrifier abundances (using nirS, nirK, and nosZ as markers) in dammed headstreams of the Nakdong River in South Korea. Sediments in Phragmites-dominated riparian areas and in-stream areas across streams (dammed vs. reference) with different streambed materials (gravel and sand) were sampled occasionally. We hypothesized that (i) the higher available N and C contents in sediments downstream of dams foster larger denitrifier communities than in the reference system and (ii) differences in potential denitrification rates across the systems correspond with denitrifier abundances. Despite 30 years of different hydrological management with dams and greater inorganic N and DOC contents in sediments downstream of dams, compared to the references, abundances of denitrifier communities and potential denitrification rates within the whole sediment were not significantly different across the systems. However, nirS and nosZ denitrifier abundances and potential denitrification rates were considerably increased in specific sediments downstream of dams (gravelly riparian and sandy in-stream) with regard to flooding events and seasonal temperature variation. nirK was not amplified in all sediments. Canonical correspondence analyses (CCA) revealed that the relationship between abundances of denitrifier communities and nutrient availabilities and potential denitrification rates was a weak one.

Comparison between Group I in Which Non-Supine Apnea-Hypopnea Index ≥5 and Group II in Which Non-Supine Apnea-Hypopnea Index<5 in Patients with Positional Sleep Apnea (체위성 폐쇄성 수면무호흡증 환자에서 비앙와위 무호흡-저호흡 지수가 5 이상과 5 미만인 집단간의 비교)

  • Park, Won Il;Jung, Hye Won;Joo, Joon Bum;Cho, Ju Eun;Kim, Jong Yang
    • Sleep Medicine and Psychophysiology
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    • v.20 no.1
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    • pp.31-34
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    • 2013
  • Objectives: The aim of this study was to evaluate the differences in patients with positional dependent sleep apnea according to their non-supine apnea-hypopnea index (AHI, ${\geq}5$ vs. <5). Methods: 92 patients with positional sleep apnea were evaluated. The patients were divided into two groups : group I was non-supine AHI having ${\geq}5$ ; group II was non-supine AHI having less than 5. Statistical analysis was performed to find the difference between two groups. Results: In 92 patients, the number of group I patients was 11 (12%) and the number of group II patients was 81 (88%). In the severe AHI group, percentage of group I was dominated (70%) and showing a significant difference compared with the mild and moderate AHI groups (p<.05). In the severe body mass index (BMI) group, percentage of group I was dominated (54.5%) and showing a significant difference compared with of the mild and moderate BMI groups (p<.05). The percentage of group I was significantly higher than group II (p<.05) in the AHI, supine AHI, non-supine AHI and snore time. Conclusions: In patients with positional sleep apnea, severe OSA and high BMI are more common in patients with non-supine AHI${\geq}5$ than non-supine AHI<5.

Clinical Effect of Bong Chuna Manual Therapy and Acupunture Treatment for Improving Cervical Curvature of Turtle Neck Syndrome and Measurement Method of Radiography (거북목증후군의 경추만곡 호전에 대한 봉추나요법과 침술의 효과와 방사선학적 평가방법 연구)

  • Oh, Won-Kyo;Lee, Eugune;Shin, Byung-Cheul
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.1
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    • pp.113-124
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    • 2009
  • Objectives : The aim of this study was to observe the effect of Bong Chuna manual therapy(BCMT) and acupuncture on the Turtle Neck Syndrome(TNS) by the changes of radiological findings and symptoms. Methods : Total twenty-eight patients diagnosed as TNS were evaluated. Twenty patients in experimental group were treated by the combination of BCMT and acupuncture, eight patients in control group were treated by acupuncture only. We measured VAS(Visual analogue scale) as pain intensity and Neck Pain and Disability Scale(NDI) at pre- and post-treatment. Also the four line Cobb's method and Jochumsen method were assessed for evaluating the radiographical changes, additionally we used 'two line method'. Results : VAS score showed statistically significant reduction in pain intensity in both experimental group and control group after 8 weeks treatment, $-44.05{\pm}14.91$ vs $-23.75{\pm}14.08$ respectively. Also NDI score presented $-11.40{\pm}8.63$ reduction in experimental group, and it was significant statistically, however not in control group, $-8.63{\pm}9.84$ reduction after 8weeks treatment. In radiological findings, the four line Cobb's method, Jochumsen method and two line method were reduced after 8weeks treatment, $-9.30{\pm}10.33$, $-0.65{\pm}1.72$ and $-14.35{\pm}5.68$ in experimental group respectively, $-0.75{\pm}1.91$, $-0.25{\pm}0.71$ and $0.38{\pm}2.20$ in control group respectively. However they were statistical significance only in the four line Cobb's method and two line method in both group. Conclusions : Combination treatment of CMT and acupuncture showed better effect on pain reduction, VAS score and the correction of neck anteversion than acupuncture only in TNS. Two line method seems valuable for evaluating the improvement of radiographical changes in TNS.

Effects of dietary supplementation of lipid-coated zinc oxide on intestinal mucosal morphology and expression of the genes associated with growth and immune function in weanling pigs

  • Song, Young Min;Kim, Myeong Hyeon;Kim, Ha Na;Jang, Insurk;Han, Jeong Hee;Fontamillas, Giselle Ann;Lee, Chul Young;Park, Byung-Chul
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.403-409
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    • 2018
  • Objective: The present study was conducted to investigate the effects of a lipid-coated zinc oxide (ZnO) supplement Shield Zn (SZ) at the sub-pharmacological concentration on intestinal morphology and gene expression in weanling pigs, with an aim to gain insights into the mechanism of actions for SZ. Methods: Forty 22-day-old weanling pigs were fed a nursery diet supplemented with 100 or 2,500 mg Zn/kg with uncoated ZnO (negative control [NC] or positive control [PC], respectively), 100, 200, or 400 mg Zn/kg with SZ for 14 days and their intestinal tissues were taken for histological and molecular biological examinations. The villus height (VH) and crypt depth (CD) of the intestinal mucosa were measured microscopically following preparation of the tissue specimen; expression of the genes associated with growth and immune function was determined using the real-time quantitative polymerase chain reaction. Results: There was no difference in daily gain, gain:feed, and diarrhea score between the SZ group and either of NC and PC. The VH and VH:CD ratio were less for the SZ group vs NC in the jejunum and duodenum, respectively (p<0.05). The jejunal mucosal mRNA levels of insulin-like growth factor (IGF-I) and interleukin (IL)-10 regressed and tended to regress (p = 0.053) on the SZ concentration with a positive coefficient, respectively, whereas the IL-6 mRNA level regressed on the SZ concentration with a negative coefficient. The mRNA levels of IGF-I, zonula occludens protein-1, tumor necrosis $factor-{\alpha}$, IL-6, and IL-10 did not differ between the SZ group and either of NC and PC; the occludin and transforming growth $factor-{\beta}1$ mRNA levels were lower for the SZ group than for PC. Conclusion: The present results are interpreted to suggest that dietary ZnO provided by SZ may play a role in intestinal mucosal growth and immune function by modulating the expression of IGF-I, IL-6, and IL-10 genes.

Impact of Low Versus Conventional Doses of Chemotherapy During Transcatheter Arterial Chemo-embolization on Serum Fibrosis Indicators and Survival of Liver Cancer Patients

  • Kong, Wei-Dong;Cao, Jian-Ming;Xu, Jian;Chen, Bo;Yang, Tao;Xu, Tan-Tan;Lu, Guang-Ming;Li, Jun;Huang, Xin-En
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4757-4761
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    • 2012
  • Objectives: To explore the impact of low- vs conventional-dose chemotherapy via transcatheter arterial chemo-embolization (TACE) on serum fibrosis indicators and treatment efficacy of hepatocellular cancer patients (HCC). Materials and Methods: Patients fulfilling the eligibility criteria were assigned to TACE in Group A (with low-dose chemotherapy) or Group B (conventional-dose chemotherapy). Four serum fibrosis related indicators, hyaluronic acid(HA), human pro-collagen type-III (hPC-III), laminin (LN), and collagen type-IV(IV-C) before TACE were compared with the values 7 days after TACE. The response rate and survival time were also compared between the two groups. Results: Fifty patients with HCC were enrolled in this study, including 25 in Group A and 25 in Group B. No significant differences were detected between the two groups in the four indicators before TACE. After TACE, the value of the four serum indicators increased significantly in Group B. However, no significant differences regarding these four indicators were found in Group A after TACE. Significant differences were demonstrated between the two groups after TACE, but median survival time and 1 or 2 year overall survival rates did not differ (P>0.05). Conclusions: Low-, compared with conventional-dose chemotherapy exerts the same impact on the variation of fibrosis related indicators and has no influence on median survival time and survival rate after TACE in HCC patients.

Cytotoxicity, Toxicity, and Anticancer Activity of Zingiber Officinale Roscoe Against Cholangiocarcinoma

  • Plengsuriyakarn, Tullayakorn;Viyanant, Vithoon;Eursitthichai, Veerachai;Tesana, Smarn;Chaijaroenkul, Wanna;Itharat, Arunporn;Na-Bangchang, Kesara
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4597-4606
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    • 2012
  • Cholangiocarcinoma (CCA) is an uncommon adenocarcinoma which arises from the epithelial cells of the bile ducts. The aim of the study was to investigate the cytotoxicity, toxicity, and anticancer activity of a crude ethanolic extract of ginger (Zingiber officinale Roscoe) against CCA. Cytotoxic activity against a CCA cell line (CL-6) was assessed by calcein-AM and Hoechst 33342 assays and anti-oxidant activity was evaluated using the DPPH assay. Investigation of apoptotic activity was performed by DNA fragmentation assay and induction of genes that may be involved in the resistance of CCA to anticancer drugs (MDR1, MRP1, MRP2, and MRP3) was examined by real-time PCR. To investigate anti-CCA activity in vivo, a total of 80 OV and nitrosamine (OV/DMN)-induced CCA hamsters were fed with the ginger extract at doses of 1000, 3000, and 5000 mg/kg body weight daily or every alternate day for 30 days. Control groups consisting of 10 hamsters for each group were fed with 5-fluorouracil (positive control) or distilled water (untreated control). Median $IC_{50}$ (concentration that inhibits cell growth by 50%) values for cytotoxicity and anti-oxidant activities of the crude ethanolic extract of ginger were 10.95, 53.15, and $27.86{\mu}g/ml$, respectively. More than ten DNA fragments were visualized and up to 7-9 fold up-regulation of MDR1 and MRP3 genes was observed following exposure to the ethanolic extract of ginger. Acute and subacute toxicity tests indicated absence of any significant toxicity at the maximum dose of 5,000 mg/kg body weight given by intragastric gavage. The survival time and survival rate of the CCA-bearing hamsters were significantly prolonged compared to the control group (median of 54 vs 17 weeks). Results from these in vitro and in vivo studies thus indicate promising anticancer activity of the crude ethanolic extract of ginger against CCA with the absence of any significant toxicity. Moreover, MDR1 and MRP3 may be involved in conferring resistance of CCA to the ginger extract.

Parental Perception on the Mechanism of Child Development and Importance of Education (속담에 나타난 아동발달 기제와 교육의 중요성에 대한 부모의 인식)

  • Lee, Joo-Yeon;Lee, Yoon Hyung
    • Korean Journal of Childcare and Education
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    • v.11 no.1
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    • pp.415-433
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    • 2015
  • The purposes of this study were to investigate Korean parents' perception on nature versus nurture and to examine the dualism in the mechanism of human development. For this goal, the present study used the Korean proverbs about child development and education as a measurement scale for developmental values. This study 1) examined the degree of agreement on the proverbs, 2) analysed the differences in the degree of agreement as the subject's socio-demographic variables, and 3) compared agreement scores as the topic of proverbs. The participants of the study were 608 parents. As the first result, the agreement scores between nature and nurture proverbs were very similar, whereas the agreement score of the importance of education proverbs was relatively higher. Second, there were some differences in related socio-economic variables as the topic of proverbs. Especially, it was revealed that the participants had some different beliefs in the nurture as their age and in the importance of education as their sex. Third, there was no significant difference between the agreement scores of nature and nurture proverbs. From these results, it seems that the interactionism between nature and nurture is generally accepted by Korean parents, and child education is a very important belief.

Influence of proton pump inhibitor therapy on intestinal inflammation assessed by fecal calprotectin in pediatric patients

  • Kim, Su Yeong;Lee, Na Mi;Yun, Sin Weon;Chae, Soo Ahn;Lim, In Seok;Choi, Eung Sang;Yi, Dae Yong
    • Clinical and Experimental Pediatrics
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    • v.62 no.10
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    • pp.400-404
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    • 2019
  • Background: An increase in the numbers of patients with gastrointestinal symptoms has recently been observed. Purpose: To investigate the effects of proton pump inhibitor (PPI) therapy on intestinal inflammation in children and adolescents as confirmed by clinical manifestations and objectively assessed by fecal calprotectin (FC) level measurement. Methods: Consecutive children (aged 3-18 years) who presented with gastrointestinal symptoms and were treated with or without PPI for at least 1 month were enrolled. Patients were divided into PPI and non-PPI groups. The PPI group was further subdivided by treatment duration and type of PPI used. Stool samples were collected for FC evaluation at baseline and after treatment and clinical data and FC levels were compared between the groups. Results: Fifty-one patients (15 boys, 36 girls) were enrolled in the study. The PPI group included 37 patients, while the non-PPI group included 14 patients. Clinical symptoms were not significantly different. FC levels and laboratory results, including C-reactive protein levels, white blood cell count, and absolute neutrophil count, were not statistically different before versus after PPI treatment. After treatment, FC levels decreased to 8.1 mg/kg (-575.4 to 340.3 mg/kg) in the PPI group and increased to 5.6 mg/kg (-460.0 to 186.9 mg/kg) in the non-PPI group compared to those before treatment (P=0.841). The number of patients with increased FC levels was not significantly different between the 2 groups (48.6% vs. 64.3%, P=0.363), similar to that observed in patients with an FC level > 50 mg/kg (24.3% and 7.1%, P=0.250). PPI therapy type and duration did not affect the FC levels (P=0.811 and P=0.502, respectively). Conclusion: Although we aimed to confirm the evidence of intestinal inflammation due to PPI use in children and adolescents through clinical symptoms and FC measurement, no significant changes were observed.