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Effects of In Vitro Fertilization Conditions of In Vitro Matured Cumulus-Intact Pig Oocytes on Embryo Development

  • Kim, Jae-Young;Lee, Eun-Ji;Park, Jin-Mo;Lee, Hong-Cheol;Park, Hum-Dai;Kim, Jae-Myeong
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.117-122
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    • 2011
  • In this study, we examined the effectiveness of in vitro fertilization of porcine immature oocytes on the embryo development of blastocysts or hatched blastocysts and the number of cells according to the in vitro fertilization conditions. In the in vitro fertilization of in vitro matured porcine oocytes, there were no significant differences between treatment groups regarding fertilization rate, blastocyst rate, and embryo development of hatched blastocysts according to the storage periods of liquid sperm of 24, 48, and 72 hours. The embryo development rate of hatched blastocysts after the fertilization according to different spermatozoa concentrations ($0.4{\times}10^5$, $1.2{\times}10^5$, and $3.6{\times}10^5$ cells/ml) showed the highest rate in the group with a spermatozoa concentration of $1.2{\times}10^5$ cells/ml; in particular, this rate was significantly higher than that in the $0.4{\times}10^5$ cells/ml group (p<0.05). The total number of blastocysts cells as well as trophectoderms (TE) that developed in each treatment group were also significantly higher in the $1.2{\times}10^5$ cells/ml group than in any other groups (p<0.05). In contrast, the embryo development rate of blastocysts according to different co-incubation periods of sperm and oocyte (1, 3, and 6 hr) was high in the 6-hour group; in particular, the rate was significantly higher than that of the I-hour group (p<0.05). Furthermore, the total number of oocytes cells and TEs that developed was significantly higher in the 6-hour group than any other group (p<0.05). In this study, the most effective treatment conditions for porcine embryo development and high cell number were found to be as follows: a sperm storage period of less than 72 hours, a spermatozoa concentration of $1.2{\times}10^5$ cells/ml, and a 6-hour co-incubation period for sperm and ooocyte.

Enhancement of Clavulanic Acid Production by Expressing Regulatory Genes in gap Gene Deletion Mutant of Streptomyces clavuligerus NRRL3585

  • Jnawali, Hum Nath;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.146-152
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    • 2010
  • Streptomyces clavuligerus NRRL3585 produces a clinically important $\beta$-lactamase inhibitor, clavulanic acid (CA). In order to increase the production of CA, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene (gap) was deleted in S. clavuligerus NRRL3585 to overcome the limited glyceraldehyde-3-phosphate pool; the replicative and integrative expressions of ccaR (specific regulator of the CA biosynthetic operon) and claR (Lys-type transcriptional activator) genes were transformed together into a deletion mutant to improve clavulanic acid production. We constructed two recombinant plasmids to enhance the production of CA in the gap1 deletion mutant of S. clavuligerus NRRL3585: pHN11 was constructed for overexpression of ccaR-claR, whereas pHN12 was constructed for their chromosomal integration. Both pHN11 and pHN12 transformants enhanced the production of CA by 2.59-fold and 5.85-fold, respectively, compared with the gap1 deletion mutant. For further enhancement of CA, we fed the pHN11 and pHN12 transformants ornithine and glycerol. Compared with the gap1 deletion mutant, ornithine increased CA production by 3.24- and 6.51-fold in the pHN11 and pHN12 transformants, respectively, glycerol increased CA by 2.96- and 6.21-fold, respectively, and ornithine and glycerol together increased CA by 3.72- and 7.02-fold, respectively.

Effects of Essential Fatty Acids during In Vitro Maturation of Porcine Oocytes: Hormone Synthesis and Embryonic Developmental Potential

  • Kim, Kang-Sig;Park, Hum-Dai
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.2
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    • pp.75-85
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    • 2019
  • Omega-3 α-linolenic acid and omega-6 linoleic acid are essential fatty acids for health maintenance of human and animals because they are not synthesized in vivo. The purpose of this study was to evaluate the effect of α-linolenic acid and linoleic acid supplementation on in vitro maturation and developmental potential of porcine oocytes. Various concentrations of α-linolenic acid and linoleic acid were added into in vitro maturation medium, and we evaluated the degree of cumulus expansion, oocyte nuclear-maturation rate, blastocyst rate, blastocyst quality, and levels of prostaglandin E2, 17β-estradiol, and progesterone in the spent medium. High doses (100 μM) of α-linolenic acid and linoleic acid supplementation significantly inhibited cumulus expansion and oocyte nuclear maturation, and prostaglandin E2 synthesis also significantly decreased compared with other groups (p < 0.05). Supplementation of 50 μM α-linolenic acid and 10 μM linoleic acid showed higher quality blastocysts in terms of high cell numbers and low apoptosis when compared with other groups (p < 0.05), and synthesis ratio of 17β-estradiol / progesterone also significantly increased compared with control group (3.59 ± 0.22 vs. 2.97 ± 0.22, 3.4 ± 0.28 vs. 2.81 ± 0.19, respectively; p < 0.05). Our results indicated that supplementation with appropriate levels of α-linolenic acid and linoleic acid beneficially affects the change of hormone synthesis (in particular, an appropriate increase in the 17β-estradiol / progesterone synthesis ratio) for controlling oocyte maturation, leading to improved embryo quality. However, high doses of α-linolenic acid and linoleic acid treatment results in detrimental effects.

Effect of High Dietary Fiber Diet on Lead Absorption and Metabolic Changes in Growing Rats (종류가 다른 고 섬유질식이가 흰쥐의 납 흡수 및 체내대사에 미치는 영향)

  • 김미경
    • Journal of Nutrition and Health
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    • v.23 no.7
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    • pp.492-503
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    • 1990
  • This study was performed to investigate nutritional effect of various dietary fibers on lead absorption, and protein and lipid metabolisms in growing rats. Sixty male rats of Sprague-Dawley strain weighing 140$\pm$1.1g were blocked into 10 groups according to body weight and fed 10 kinds of diet different with fiber sources [non-fiber, cellulose, pectin, guar gum or carboxymethylcellulose(CMC)] and lead levels (0 or 1%) for 4 weeks. Results were summerized as follows : 1) Food intake, weight gain, FER and PER were remarkably decreased in lead(Pb)-added groups. Weight gain, FER and PER in Pb-added pectin group were significantly lower than those in Pb-added non-fiber group. 2) Liver and kidney weights, femur weight and length, hematocrit and hemoglobin content were decreased in Pb-added groups. Especially femur and liver weights in pectin groups were the lowest among groups. 3) Total protein content in serum was significantly decreased in Pb-added groups but was not different with dietary fiber sources. Total lipid content in serum was not different with dietary Pb levels and fiber sources, but cholesterol content in serum of guar gum group was significantly decreased by Pb addition. 4) Nitrogen, lipid and cholesteol contents in liver were significantly decreased in Pb-added groups, and lipid content in liver of pectin and CMC groups was lower than other groups. 5) Daily urinary and fecal excretions of nitrogen, kipid and cholesterol were decreased in Pb-added groups, and fecal nitrogen was significantly increased in Pb-added groups, and fecal nitrogen of cellulose and guar gum groups was significantly higher than other groups. Fecal excretions of lipid and cholesterol were increased by dietary fibers, and especially fecal lipid was remarkably increased in pectin and guar hum group. 6) Pb contents in liver and femur were decreased by dietary fibers. Especially Pb contents in liver, kidney and femur were significantly decreased in guar gum group. 7) Daily urinary and fecal excretions of Pb were significantly increased in cellulose and guar gum groups, and fecla excretion of Pb in guar gum group was twice of non-fiber group. Pb absorption ratio was significantly decreased in guar gum group. In conclusion, dietary fibers have effect on protein and lipid metabolisms, and decreased intestinal absorption of Pb by increasing fecal excretion. But the degree of effect was different with dietary fiber sources.

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Perioperative Risk Factors Related to Lumbar Spine Fusion Surgery in Korean Geriatric Patients

  • Lee, Jung-Hyun;Chun, Hyoung-Joon;Yi, Hyeong-Joong;Bak, Koang-Hum;Ko, Yong;Lee, Yoon-Kyoung
    • Journal of Korean Neurosurgical Society
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    • v.51 no.6
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    • pp.350-358
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    • 2012
  • Objective : Life expectancy for humans has increased dramatically and with this there has been a considerable increase in the number of patients suffering from lumbar spine disease. Symptomatic lumbar spinal disease should be treated, even in the elderly, and surgical procedures such as fusion surgery are needed for moderate to severe lumbar spinal disease. However, various perioperative complications are associated with fusion surgery. The aim of this study was to examine perioperative complications and assess risk factors associated with lumbar spinal fusion, focusing on geriatric patients at least 70 years of age in the Republic of Korea. Methods : We retrospectively investigated 489 patients with various lumbar spinal diseases who underwent lumbar spinal fusion surgery between 2003 and 2007 at our institution. Three fusion procedures and the number of fused segments were analyzed in this study. Chronic diseases were also evaluated. Risk factors for complications and their association with age were analyzed. Results : In this study, 74 patients experienced complications (15%). The rate of perioperative complications was significantly higher in patients 70 years of age or older than in other age groups (univariate analysis, p=0.001; multivariate analysis, p=0.004). However, perioperative complications were not significantly associated with the other factors tested (sex, comorbidities, operation procedures, fusion segments involved). Conclusion : Increasing age was an important risk factor for perioperative complications in patients undergoing lumbar spinal fusion surgery whereas other factors were not significant. We recommend good clinical judgment and careful selection of geriatric patients undergoing lumbar spinal fusion surgery.

A Study on Cognition of Oriental Medical Hospital Employees on the Oriental Medical Institution Assessment (한방의료기관평가에 대한 한방의료기관 근무자의 인지도)

  • Jeon, Hyun-Sook;Jung, Sang-Hyuk;Yu, Seung-Hum;Jung, Woo-Sang
    • The Journal of Korean Medicine
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    • v.29 no.4
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    • pp.114-122
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    • 2008
  • Objectives: To determine the factors affecting the assessment of Oriental medical institutions under the Oriental Medical Institution Assessment System and thereupon, provide for some basic data and alternative measures for assessment of Oriental medical institutions. Methods: The researcher sampled 320 people employed by 26 Oriental medical institutions designated as model Oriental medical institutions subject to assessment in 2008 and 2009 and thereupon, used a structured and open-ended survey table for them to collect the data. The size of the final sample was n=302. Results: The Oriental medical institution staff were highly aware of the Oriental medical institution assessment (OMIA), After adjusting the other factors by multiple regression, the factors affecting such recognition were different significantly depending on age (those in their 30's), types of job (nurses and treatment assistants) and locations of hospital (GyeongSang-do). The staff expected the OMIA could helpful for improving facilities and system of hospitals, thus promoting satisfaction of patients. To do so, they felt it necessary to develop an assessment scale reflecting the special conditions besetting the Oriental medical institutions as well as the indices for improvement of Oriental medical service quality. Conclusion: It is hoped that this study will be followed up by future studies which will comparatively analyze Oriental medical institution staff's perception of the assessment system before and after its operation and thereby, suggest some ideal policy alternatives for assessment of the Oriental medical institutions. Furthermore, future studies are requested to research into Oriental medical institution staff's needs and consumers' needs as well in consideration of the characteristics of the Oriental medical institutions and thereupon, suggest some alternatives for continued education, development of the assessment tools, methods and policies.

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Development of a Critical Pathway of Bullectomy for Spontaneous Pneumothorax Patients (자연기흉으로 폐기포절제술을 받은 환자를 위한 표준진료지침서 개발)

  • Kim, Mi Kyoung;Yu, Seung Hum;Lee, Doo Yun;Son, Young Mo
    • Quality Improvement in Health Care
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    • v.9 no.1
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    • pp.34-51
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    • 2002
  • Background : The purpose for this study is to develop a critical pathway of bullectomy for spontaneous pneumothorax patients. Methods : For this study a conceptual framework of critical pathway was developed through a review of the literature including five critical pathways which are currently being used in USA, and opinions of the critical pathway development team members at Y university hospital. In order to identify the service contents required by these patients and to draw up a preliminary critical pathway, 33 cases of medical records of patients who had received bullectomy for spontaneous pneumothorax between September, 2000 to August, 2001 at the Respiratory Center of Y university hospital in Seoul was analyzed. Results : In order to test the clinical validity of the preliminary critical pathway, it was applied to ten patients who had received bullectomy for spontaneous pneumothorax from October, 2001 to December, 2001. The average discharge day was 4.89th post operation day, six patients discharged on the fourth post operation day which was the expected day, one patients discharged one day earlier than the expected day, one patient discharged three days later than the expected day, and one patient discharged six days later than the expected day. There were variances between the critical pathway and the actual practice. The variances came from tests, medications, and treatments. One item that showed variance in clinical applications was complemented, and three items were decided not to be corrected for the final determination of the critical pathway. Conclusion : This critical pathway is applicable to the care of patients with bullectomy for spontaneous pneumothorax, but it needs more clinical applications to grasp varied variances.

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Surgical Experience of Transsphenoidal-Supradiaphragmatic Intradural Approach to Presellar and Suprasellar Lesions

  • Park, Min-Woo;Kim, Jae-Min;Kim, Jae-Hoon;Bak, Koang-Hum;Kim, Choong-Hyun;Jeong, Jin-Hyeok
    • Journal of Korean Neurosurgical Society
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    • v.39 no.5
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    • pp.329-334
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    • 2006
  • Objective : In selected cases, the transsphenoidal approach[TSA] can be extended anteriorly to the tuberculum sellae, chiasmatic sulcus, and planum sphenoidale to obtain direct exposure of the suprasellar cisterns and its contents. We applied this modification of the TSA to various lesions of the presellar and suprasellar areas. We evaluate our clinical experience of this technique and review the related literature. Methods : From 1999 to 2004, we used the transsphenoidal supradiaphragmatic intradural approachs[TSIAs] in 9 patients who had various lesions at the pre- and suprasellar regions. Concomitant presellar extension of the bone window was performed with the sublabial or transnasal transseptal transphenoidal techniques. After removal of the lesions, sellar or anterior cranial floor was repaired with silicone plate substitute. Results : The TSIAs have been applied in the following cases : four tuberculum sellae meningiomas, two craniopharyngiomas, two Rathke's cleft cysts, and one non-functioning macroadenoma. The complications were one case of visual acuity decrease and one cerebrospinal fluid rhinorrhea. Conclusion : The TSIA is easily applicable through a minor modification of the standard TSA. It is suitable for removing lesions located in the presellar and suprasellar area adjacent to the pituitary stalk with minimal brain manipulation and decreased morbidity.

Efficacy of the Decompressive Craniectomy for Acute Cerebral Infarction : Timing of Surgical Intervention and Clinical Prognostic Factors

  • Cho, Tae-Koo;Cheong, Jin-Hwan;Kim, Jae-Hoon;Bak, Koang-Hum;Kim, Choong-Hyun;Kim, Jae-Min
    • Journal of Korean Neurosurgical Society
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    • v.40 no.1
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    • pp.11-15
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    • 2006
  • Objective : Acute cerebral infarction is often accompanied by transtentorial herniation which can be fatal. The aim of this study is to determine the timing of surgical intervention and prognostic factors in patients who present with acute cerebral infarction. Methods : We reviewed retrospectively 23 patients with acute cerebral infarction, who received decompressive craniectomy or conservative treatment from January 2002 to December 2004. We divided patients into two groups according to the treatment modalities [Group 1 : conservative treatment, Group 2 : decompressive craniectomy]. In all patients, the outcome was quantified with Glasgow Outcome Scale and Barthel Index. Results : Of the 23 patients, 11 underwent decompressive craniectomy. With decompressive craniectomy at the time of loss of pupillary light reflex, we were able to prevent death secondary to severe brain edema in all cases. Preoperative Glasgow Coma Scale and loss of pupillary light reflex were significant to the clinical outcome statistically. With conservative treatment, 9 of the 12 patients died secondary to transtentorial herniation. The clinical outcomes of remaining 3 patients were poor. Conclusion : This study confirms the value of life-saving procedure of decompressive craniectomy after acute cerebral infarction. We propose that the loss of pupillary light reflex should be considered one of the most important factors to determine the timing of the decompressive craniectomy.

Co-expression of Survivin and Bcl-2 in Primary Brain Tumors : Their Potential Effect on Anti-apoptosis

  • Ryu, Je-Il;Kim, Choong-Hyun;Cheong, Jin-Hwan;Bak, Koang-Hum;Kim, Jae-Min;Oh, Suck-Jun
    • Journal of Korean Neurosurgical Society
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    • v.40 no.1
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    • pp.1-5
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    • 2006
  • Objective : Survivin is an inhibitor of apoptosis protein[IAP], which inhibits apoptosis through a pathway distinct from the Bcl-2 family members. Overexpression of survivin and Bcl-2 have been commonly reported in human neoplasms. The authors investigate whether there is a synergistic effect on the anti-apoptosis rate of primary brain tumors "in situ" based on the co-expression of survivin and Bcl-2. Methods : One hundred and two brain tumor patients who had been resected were included in this study. Survivin tin and Bcl-2 were detected by Western blotting analysis, while apoptosis was examined by DNA fragmentation analysis. An anti-apoptotic rate was assessed in these brain tumor samples based on the expression of survivin and Bcl-2 or co-expression of both. Results : Survivin and Bcl-2 were expressed in 57[55.9%] and 53[52.0%] of 102 brain tumor samples studied respectively, and co-expressed in 31[30.4%]. The percentage of astrocytic and meningeal tumors expressing survivin was significantly correlated with histological grades; however, Bcl-2 was not correlated [p=0.106]. The anti-apoptotic rate in primary brain tumors with survivin, Bcl-2, and both was detected in 49[86.0%] of 57 samples, 42[79.9%] of 53 samples, and 27[87.1%] of 31 samples, respectively. Their difference in the frequency of anti-apoptosis was not significant. Conclusion : Survivin or Bcl-2 is involved in the anti-apoptosis. However, it suggests that co-expression of survivin and Bcl-2, together, have no synergistic effect on the anti-apoptotic properties of the primary brain tumors.