Kim, Soo-Kyung;Kang, Hee-Kyoo;Jun, Jin-Hyun;Choi, Kyoo-Wan;Kim, Moon-Kyoo
Development and Reproduction
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v.5
no.1
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pp.17-21
/
2001
This study was conducted to investigate the expression pattern of Trypsin-like enzyme and the effect of a trypsin inhibitor(benzimidine) on hatching process during in-vitro culture of mouse preimplantation embryos. The Trypsin-like enzyme was identified by rhodamine-conjugated Trypsin substrate probe. The expression of trypsin-like enzyme was firstly detected at the late morula stage, and the enzyme was uniformly localized in the trophectoderm of late blastocysts. Especially, intense fluorescence was observed in the blebbing area of hatching blastocysts. Bisbenzamidine, contained in culture media, did not alter embryonic development from 4-cell stage to the expanded blastocyst but decrease the hatching rate in ImM concentration (15.8% vs 89.7%, p<0.02). In the treatment of bisbenzimidine (5mM) for 12 hours according to the embryonic stage of mouse, the hatching rate of control (83.0%) and treatment in late blastocysts (8.7%) were significantly (p<0.01) different. From these results, we suggested that the hatching enzyme having trypsin-like activity was localized from the late morula stage, and the hatching process by this enzyme was activated in the late blastocyst stage of mouse embryos.
Kim, Joung-Hee;Kwack, Seung Jun;Cho, Yong-Kwon;Kim, Byung-Kwan;Kim, Jong Guk;Lee, Eunkyung;Cho, Hye-Kyung;Kim, Keuk-Jun
Journal of Life Science
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v.25
no.7
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pp.748-756
/
2015
The report explores the possibility that the single extract of natural substances and the mixture of single extracts of natural substances create a synergy effect to increase the antimicrobial activity. It also compares the previous researches to find out if the natural medicinal herbs’ extract has antimicrobial activity on dandruff causative organism, Malassezia furfur. For the experiment on the mixture of single extracts of natural substances, the results are like following: 1. Staphylococcus aures’s antibacterial activity is resistant to mixture of three natural substances. 2. Escherichia coil’s antibacterial activity is resistant to mixture with Coptidis rhizoma. 3. Candida albicans’ antifungal activity is resistant to mixture with Chinese galls, which means the different results may be expected when tested with each germ. 4. On the other hand, Malassezia furfur has no antifungal activity on the single extract of natural substances and shows weak antifungal activity, whose diameter is 3.20 mm when tested with the mixture of 50% of Coptidis rhizoma and 50% of Phytoncide. The result is totally different from the one on the same eumycetes, C. albicans. That is because M. furfur has more lipophilic chemicaled cell walls than C. albicans does and it also consists of lamella layer, inner plasma membrane and intermediate multiple layers.
Uncoupling protein (UCP) 3 has a number of proposed roles in the regulation of fatty acid metabolism. A number of polymorphisms in the human UCP3 gene have been identified, and the correlation with obesity related phenotypes evaluated. The objective of this study was to identify SNP in porcine UCP3 gene and to investigate the effect of the SNP on economic traits. The sequencing analysis method was used to identify nucleotide polymorphisms at position 1405 bp (Genebank accession No : AY739704) in porcine UCP3 gene. The SNP (G150R), located in the exon 3, changed the amino acid to glycine (GGG) from arginine (AGG). This G150R showed three genotypes - GG, GR and RR - by digestion with the restriction enzyme Sma Ⅰ using the PCR-RFLP method. The G150R showed significant effects only on back fat (P<0.05). Animals with the genotype GG had significantly higher back fat thickness (1.358 cm) than animals with the genotype GR (1.288 cm, P<0.05) and RR (1.286 cm, P<0.05). However, the genotypes had no significant association with ADG and days to 90kg. According to results of this study, a G allele of the G150R was found to have a significant effect on back fat thickness. It will be possible to use SNP markers on selected pigs to improve backfat thickness, an important economic trait.
Mobilization of storage lipids is critical for the germination of oil seeds, as they supply carbon and energy until photosynthesis commences in cotyledons. In this study, we determined the levels of plant carnitine and associated changes in these levels from seed germination to cotyledon senescence. We also examined changes in the content of unsaturated fatty acids throughout seedling development. Carnitine levels peaked on day 3 at 14.5 nM in cotyledons and decreased sharply to 7.2 nM on day 4. On development day 3 carnitine levels were maintained at around 3 nM until day 7. The unsaturated fatty acid content dropped by half at the same time as carnitine peaked (day-3), and storage lipids were almost depleted by day 5. Thereafter, carnitine was hardly detected until the second stage of cotyledon senescence, at which stage the carnitine content was 6.8 nM, similar to that on day 4 at the time of fatty acid depletion in the cotyledons. Unsaturated fatty acids levels remained constant in green cotyledons but slightly increased in the senescing cotyledons. The latter can be explained by intracellular breakdown of membrane lipids. This is the first such discovery in developing cotyledons and may offer clues regarding other roles of the acetyl unit transport system in plants. The expression of BOU was closely associated with carnitine metabolism during seed germination and cotyledon development. The results provide support for the possibility of carbon re-routing during the glyoxylate cycle in the supply of energy for early germination and development.
Membrane proteins are involved in many common diseases, including heart disease and cancer. In various disease states, such as cancer, abnormal signaling pathways that are related to the membrane proteins cause the cells to divide out of control and the expression of membrane proteins can be altered. Membrane proteins have the hydrophobic environment of a lipid bilayer, which makes an analysis of the membrane proteins notoriously difficult. Therefore, this study evaluated the efficacy of two different methods for optimal membrane protein extraction. High-speed centrifuge and reagent-based method with a -/+ filter aided sample preparation (FASP) were compared. As a result, the high-speed centrifuge method is quite effective in analyzing the mitochondrial inner membranes, while the reagent-based method is useful for endoplasmic reticulum membrane analysis. In addition, the function of the membrane proteins extracted from the two methods were analyzed using GeneGo software. GO processes showed that the endoplasmic reticulum-related responses had higher significance in the reagent-based method. An analysis of the process networks showed that one cluster in the high-speed centrifuge method and four clusters in the reagent-based method were visualized. In conclusion, the two methods are useful for the analysis of different subcellular membrane proteins, and are expected to assist in selecting the membrane protein extraction method by considering the target subcellular membrane proteins for study.
There is growing concern that dietary soy intake is associated with protection of breast cancer. However, questions persist on the potential adverse effects of the main soy constituent genistein(GS) on female reproductive physiology. In this study, we examined whether prepubertal exposure to GS affected on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. GS(100mg/kg/day) was administrated daily from postnatal day 25(PND 25) to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the day after VO occurred. Gross anatomy and tissue weight were compared to test the GS's effect on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in tissues. Specific radioimmunoassay(RIA) were carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptors(PR), total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a results, advanced VO was shown in the GS group(PND $31.2{\pm}0.6$) compared to the vehicle group (PND $35.3{\pm}0.7$). GS treatment significantly increased wet weight of ovaries and uteri compared to the vehicle group. Increased serum LH levels were also shown in the GS group. Graafian follicles and corpora lutea(CL) were observed only in the ovaries from GS treated animals. Similarly, hypertrophy of luminal and glandular uterine epithelium were found only in the GS group. Collectively, these effects were probably due to the estrogenic effects of GS. In the semi-quantitative RT-PCR studies, the transcriptional activities of PR in both ovary and uterus from GS-treated group were significantly higher than those from the vehicle group. The present studies demonstrated that acute exposure to GS, at levels comparable to the ranges of human exposure, during the critical period of prepubertal stage activates the reproductive system resulting precocious puberty in immature female rats.
The present study was designed to examine effect of short term treadmill and weight-training on aging arophy in the rat skeletal muscle. Male rats of 24 months old were used. Each groups included control, treadmill and weight-training for 4 weeks by using treadmill apparatus and body press apparatus. The histo and cytochemical, ultrastructural and stereological changes in senile skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups remained constant. The volume density of muscle fiber type IIC and IIB were increased, that of type IIA was decreased, but type I remained constant in treadmill-training group. In weight-training rat, the density of type IIA and IIB were increased, both those of type IIC was decreased. But, all changes of muscle fiber type is not significant. Senile control group some usual formation of mild contraction band, liposuscin pigment and muscular splitting were observed. After treadmill-training, histological and ultrastructural changes occurred in the muscle fiber, such as irregularity of the sarcolemma, interfibrillar vacuolization, longitudinal splitting, and widened I-bond. After weight-training, the changes occurred in the trained muscle fiber, such as appearances of many lysosomes and autophagic vacuoles, severe contraction band, and breakup of myofibrils. Histo and cytochemical studies showed that the activities of succinic dehydrogenase and acid phosphatase remained constant, activities of $Mg^{++}$-ATPase decrease with training. Stereological changes were not observed in the volume and numerical density of all subject component, but the surface density of mitochondrial inner membrane was increased with treadmill-training. These experimental results suggested that endurance training during short-term may result in the adaptible response in senile skeletal muscles. On the other side, weight-training is bad for senile skeletal muscle.
Vinclozolin(VCZ), a systemic dicarboximide fungicide, has been used in the control of diseases caused by microorganism of some species in fruits, vegatables and ornamental plants. Although VCZ itself is a very weak antagonist for androgen receptor binding, both melabolites M1 and M2 are effective antagonists. The present study was undertaken to examine whether prepubertal exposure to VCZ affects on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. VCZ(10 mg/kg/day) was administered daily from postnatal day 21(PND 21) through the day when the first vaginal opening(V.O.) was observed. Gross anatomy and weight of reproductive tissues were compared to test the VCZ's effects on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in the tissues. To determine the transcriptional changes in progesterone receptor(PR), total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a result, delayed V.O. was shown in the VCZ group(PND $34.00{\pm}1.22$) compared to the control group(PND $38.20{\pm}1.92$; p<0.01). VCZ treatment significantly decreased the wet weight of ovaries and uteri compared to the control group(p<0.01). Graafian follicles and corpora lutea were observed only in the ovaries from the control animals, while numerous primary, secondary follicles and small atretic follicles were observed in the ovaries from VCZ group. Similarly, hypotrophy of luminal and glandular uterine epithelium was found in the VCZ group. In the semi-quantitative RT-PCR studies, the transcriptional activity of PR in ovary(p<0.01) from VCZ group were significantly lower than those from the control group while in uterus were similar compared with the control group. The present studies demonstrated that the acute exposure to VCZ during the critical period of prepubertal stage could inactivate the reproductive system resulting delayed puberty in female rats.
Kim Jae-Hyun;Oh Sam-Sae;Lee Chang-Ha;Baek Man-Jong;Hwang Seong-Wook;Lee Cheul;Lim Hong-Gook;Na Chan-Young
Journal of Chest Surgery
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v.39
no.4
s.261
/
pp.289-297
/
2006
Background: Surgery for Stanford type A aortic dissection shows a high operative mortality rate and frequent postoperative brain injury. This study was designed to find out the risk factors leading to operative mortality and brain injury after surgical repair in patients with type A aortic dissection. Material and Method: One hundred and eleven patients with type A aortic dissection who underwent surgical repair between February, 1995 and January 2005 were reviewed retrospectively. There were 99 acute dissections and 12 chronic dissections. Univariate and multivariate analysis were performed to identify risk factors of operative mortality and brain injury. Resuit: Hospital mortality occurred in 6 patients (5.4%). Permanent neurologic deficit occurred in 8 patients (7.2%) and transient neurologic deficit in 4 (3.6%). Overall 1, 5, 7 year survival rate was 94.4, 86.3, and 81.5%, respectively. Univariate analysis revealed 4 risk factors to be statistically significant as predictors of mortality: previous chronic type III dissection, emergency operation, intimal tear in aortic arch, and deep hypothemic circulatory arrest (DHCA) for more than 45 minutes. Multivariate analysis revealed previous chronic type III aortic dissection (odds ratio (OR) 52.2), and DHCA for more than 45 minutes (OR 12.0) as risk factors of operative mortality. Pathological obesity (OR 12.9) and total arch replacement (OR 8.5) were statistically significant risk factors of brain injury in multivariate analysis. Conclusion: The result of surgical repair for Stanford type A aortic dissection was good when we took into account the mortality rate, the incidence of neurologic injury, and the long-term survival rate. Surgery of type A aortic dissection in patients with a history of chronic type III dissection may increase the risk of operative mortality. Special care should be taken and efforts to reduce the hypothermic circulatory arrest time should alway: be kept in mind. Surgeons who are planning to operate on patients with pathological obesity, or total arch replacement should be seriously consider for there is a higher risk of brain injury.
Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
Journal of Embryo Transfer
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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.
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