• Journal of Embryo Transfer
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• v.17 no.3
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• pp.239-249
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• 2002

Embryos derived from pig oocytes matured in mSOF are able to develop to blastocysts after IVF. Experiment 1 evaluated the effects of two maturation media (TCM-199 vs mSOF) on maturation rate, fertilization parameters, including penetration, polyspermy, male pronuclear formation, and the mean number of sperm penetrated per oocyte. Experiment 2 and Experiments 3 examined the effects of two maturation media on zona pellucida solubility and cortical granule distribution by transmissible electron microscopy, respectively. Experiment 4 assessed the effects of two maturation media on the in vitro embryo cleavage rate and development to blastocyst. Lastly, experiment 5 examined the cell number of blastocyst. An effect of media (P<0.05) was detected for mSOF on the mean number of sperm per oocyte. In TCM group, zona digestion time (196.5$\pm$15.5 vs 131.6$\pm$20.1 before IVF, 397.5$\pm$30.3s vs 185.3$\pm$16.4s after IVF, p<0.05) was higher in TCM-199 group. No significant effects of media was observed on cortical granule distribution between two groups by TEM. An effect (P<0.05) was observed on embryo development to blastocyst (16% vs 8%) but not on cleavage rates. No significant effects of media was observed on total cell number of blastocyst. We found that the high mean number of sperm penetrated per oocyte and the weaker zona pellucida on the basis of the digestion time was shown in pig oocytes matured in mSOF, however, porcine oocyte maturation with supplemented synthetic oviduct fluid medium (mSOF) resulted in blastocyst cell numbers comparable to those observed with Tissue Culture Medium 199.

• Biomolecules & Therapeutics
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• v.15 no.4
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• pp.205-211
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• 2007

The fruits and stems of Opuntia ficus-indica var. saboten have been reported to exhibit a variety of pharmacological actions, including antioxidant, analgesic, anti-inflammatory, and anti-ulcer effects. In the present study, we evaluated effects of the butanol fraction (SK OFB901) prepared from the 50% ethanol extract of the stems on various types of neuronal injuries induced by oxidative stress, excitotoxins, and amyloid ${\beta}\;(A_{\beta})$ in primary cultured rat cortical cells. Its antioxidant and radical scavenging activities were also evaluated by cell-free bioassays. We found that SK OFB901 strongly inhibited the oxidative neuronal damage induced by $H_2O_2$ or xanthine/xanthine oxidase. In addition, it exhibited marked inhibition of the excitotoxic neuronal damage induced by glutamate, N-methyl-D-aspartic acid, or kainate. Furthermore, the $A_{\beta(25-35)}$-induced neurotoxicity was also significantly attenuated by SK OFB901. It was found to inhibit lipid peroxidation initiated by $Fe^{2+}$ and L-ascorbic acid in rat brain homogenates and scavenge 1,1-diphenyl-2-picrylhydrazyl free radicals. These results indicate that the butanol fraction prepared from the stems of Opuntia ficus-indica var. saboten exerts potent antioxidant and neuroprotective effects through multiple mechanisms, implying its potential applications for the prevention or management of neurodegenerative disorders associated with oxidative stress, excitotoxicity, and $A{\beta}$.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.183-191
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• 1993

These experiments were carried out to investigate whether the enzyme is involved in zona hardening during normal activatin of the oocytes by sperm, and demonstrate peroxidase activity during in vitro fertilization of oocytes treated with peroxidase inhibitors(250 $\mu$M phenylhydrazine, 28mM sodium sulfite, 350mM glycine ethyl ester and 50mM sodium azide) and tyrosine analogue(12.5mM tyramine). Also, zona soluble properties of the ovarian oocytes incubated for 0, 5, 10 and 15 hr in the presence of pheylhydrazine or tyramine were studied by using $\alpha$-chymotrypsin. The results obtained from these experiments were summarized as follows ; 1. The rates of fertilizatin in control oocytes and oocytes treated with phenylhydrazine or tyramine were 69.8%, 62.3% and 88.2%, respectively. However in vitro fertilization in oocytes treated with three different peroxidase inhibitors, sodium sulfite, glycine ethyl ester and sodium azide, were not induced. The oocytes treated with phenylhydrazine had no significant effect on in vitro fertilization rate as compared to control. However there was a significantly different in fertilization between tyramine treated group and control group(P<0.01). 2. The zona solubility(t50) of control and fertilized oocytes in culture treated with phenylhydrazine or tyramine were 30.7, 26.0 and 16.3 min., respectively. Phenylhydrazine treated group and tyramine treated group had effect on inhibition of zona hardening as compared to control group. These results suggest that ovoperoxidase is involved in zona hardening during normal activation of the oocytes by sperm. 3. t50 of control oocytes and ovarian oocytes treated with phenylhydrazine or tyramine for 5, 10 and 15 hr in vitro were 14.0, 26.2 and 32.0 min., 14.5, 26.9 and 30.2 min., and 14.0, 24.3 and 31.2 min., respectively. These results suggest that zona hardening in ovarian oocytes matured for various times in vitro cannot be inhibited by peroxidase inhibitors and tyrosine analogue, that the spontaneous zona hardening incultured ovarian oocytes is not caused by the secretory products of cortical granules released during the cortical reaction, ovoperoxidase.

• Journal of Life Science
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• v.17 no.1 s.81
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• pp.143-149
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• 2007

Scutellaria baicalensis GEORGI(SB) is used in oriental medicine for the treatment of incipient strokes. Although it has been reported that SB is neuroprotective in a hypoxia model, its mechanism is poorly understood. Here, we investigated the effect of SB on the modulation of retinoic acid receptor a (RARa). Rat cerebrocortical cells were grown in neurobasal medium. On DIV12 cells were treated with SB $(20{\mu}g/ml)$ and given a hypoxic shock $(2%\;O_2/5%\;CO_2,\;3hr)$ on DIV14. In situ hybridization using cRNA probe revealed that RARa mRNA punctae are distributed, in addition to nucleus, throughout neuronal dendrites, where SB upregulated its density by 69.8% (p=0.001) and 129.8% (p=0.001) in both normoxia and hypoxia, respectively. At the protein level, SB upregulated RARa in the neuronal soma by 78.8% (p=0.004) and 23.6% (p=0.001) in both normoxia and hypoxia, respectively. These results indicate that SB upregulates RARa in both normoxia and hypoxia, which might contribute to the neuroprotection.

• Journal of Plant Biology
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• v.36 no.2
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• pp.177-182
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• 1993

The root nodules of Elaeagnus umbellata were coralloid-shape due to repeated dichotomous branching of nodule meristem. The filamentous endophyte with vesicle cluster ranging from 30 ${\mu}{\textrm}{m}$ to 60 ${\mu}{\textrm}{m}$ in diameter was present only in the cortical cells. The isolated endophytes in vitro culture showed typical Frankia morphology, consisting of highly branched hyphae ranging from 0.8 ${\mu}{\textrm}{m}$ to 1.0 ${\mu}{\textrm}{m}$ in diameter, terminal and intrahyphal sporangia varing in shape and size up to 60 ${\mu}{\textrm}{m}$ in length and laminated vesicles. Its infectivity and effectivity were confirmed by induction of nitrogen-fixing root nodules on the inoculated seedlings of two Elaeagnus species. Consequently, the isolate was confirmed as a true symbiont of Elaeagnus umbellata root nodule and named Frankia EuIK1.

• The Journal of Internal Korean Medicine
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• v.25 no.4
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• pp.324-336
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• 2004

Objectives : The purpose of this investigation is to evaluate the effects of Scutellaria baicalensis GEORGI on alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E18 rat cortical cells were grown in a Neurobasal medium containing B27 supplement. On 12 DIV, Scutellaria baicalensis GEORGI(20 ug/ml) was added to the culture media and left for 24 hrs. On 11 DIV, cells were given a hypoxic insult $(2%\;O_2/5%\;CO_2,\;37^{\circ}C,\;3\;hrs)$, returned to normoxia and cultured for another 24 hrs. Total RNA was prepared from Scutellaria baicalensis GEORGI-untreated (control) and -treated cultures and alteration in gene expression was analysed by microarray using rat 5K-TwinChips. Results : For most of the genes altered in expression, the Global M values were between -0.5 to +0.5. Among these, 1143 genes increased in their expression by more than Global M +0.1, while 1161 genes decreased by more than Global M -0.1. Effects on some of the genes whose functions are implicated in neural viability are as follows: 1) The expression of apoptosis-related genes such as Bad (Global M = 0.39), programmed cell death-2(Pdcd2) (Global M = 0.20) increased, while Purinergic receptor P2X(P2rxl) Global M = -0.22), Bc12-like1(Bc1211)(Global M = -0.19) decreased. 2) The expression of 'response to stress-related genes such as antioxidation-related AMP-activated protein kinase subunit gamma 1 gene (Prkag1) (Global M = 0.14), catalase gene (Global M = 0.14) and Heme Oxygenase(Hmoxl) increased. 3) The expression of Fos like antigen 2 (Fos12) expressed in neurons that survive ischemic insult increased (Global M = 0.97). Conclusions : these data suggest that Scutellaria baicalensis GEORGI increases the expression of antiapoptosis- and antioxidation- related genes in a way that can not yet be explained.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.600-606
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• 2005

Neurofilament (NF) proteins constitute the major intermediate filament type in adult neurons. They are made up by the copolymerization of the neurofilament light (NF-L, 61 kDa), medium (NF-M, 90kDa), and heavy (NF-H, 115 kDa) proteins. Although neurofilaments play a crucial .ole in neuronal growth, organization, shape, and plasticity, their expression pattern and cellular distribution in the developing neurons remain unknown. In this study, we have produced a rabbit polyclonal antibody specific to NF-M and investigated expression of NF-M in cultured cortical neurons. Immunostaining of 12 and 24 h cultures revealed strong expression of NF-M in axonal growth cone and in the region of a soma toward the axon. Doublestaining of 4 and 14 DIV corical neurons with NF-M and PSD95 antibodies revealed that both axon and dendrites were stained intensely with NF-M antibody, and that NF-M immunostaining along dendrites is often punctate and colocalize with PSD95 puncta, indicating that the puncta represent postsynaptic spines. Presence of NF-M in the postsynaptic spine was also indicated by immunoblot analysis of the postsynaptic density fraction. Taken together, our results show intensive targeting of NF-M into axons in the early axonal development, and into spines in mature neurons, indicating its important functions in axon and spine development.

• Childhood Kidney Diseases
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• v.14 no.1
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• pp.71-78
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• 2010

Purpose : Hydronephrosis is found about 30% of children with urinary tract infection (UTI). It can be caused by various conditions, although most childhood hydronephrosis is congenital. This study was performed to investigate the relationship between febrile UTI and hydronephrosis. Methods : We retrospectively reviewed the medical charts of 183 patients diagnosed as UTI between January 2007 and May 2009 at Korea University Guro Hospital. Inclusion criteria were as followings; 1) fever more than $37.5^{\circ}C$ measured in the axilla, 2) positive urine culture, 3) no history of urinary tract anomaly on antenatal sonography and urinary tract infection. We classified the enrolled children into two groups of patients with hydronephrosis (HN) and those without hydronephrosis (NHN). Results : The 80 patients were HN and 103 patients NHN. Hydronephrosis was found in 58 patients with left kidney, 8 right and 14 both kidneys. Most of hydronephrosis were of low grade. Compared with NHN group, initial renal cortical defects on DMSA scan significantly increased in HN group (HN 37.5%, NHN 16.5%, P<0.05). The incidence of VUR was not different between the two groups (HN 22%, NHN 12.1%). White blood cell counts and C-reactive protein were not different between the two groups. Follow-up DMSA scan (about 6 months later after UTI) showed no difference of renal scarring in both two groups. Conclusion : Our data suggests that hydronephrosis in febrile UTI patients is clinically useful for detecting renal cortical defects, but is not associated with follow-up renal scar.

• Clinical and Experimental Pediatrics
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• v.46 no.11
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• pp.1101-1106
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• 2003

Purpose : In vivo, minocycline appears to be neuroprotective. Thus, the neuroprotective effects of minocycline were studied in a rat brain cortical cell culture induced by hypoxia. Methods : Cultured cells from the brains of Sprague-Dawley rats were divided into two sets of groups : normoxia groups treated with 5% $CO_2$ and hypoxia groups treated with 1% $CO_2$. After several days of incubation, the control groups were not treated with minocycline, while the sample groups were treated with either 1 or $10{\mu}g/mL$ of minocycline. The damaged cells were observed under a microscope, while apoptosis was detected using a TUNEL assay control-stained with DAPI. Results : Among the normoxia groups, the control and sample groups treated with 1 and $10{\mu}g/mL$ of minocycline were all statistically significantly different from each other. Meanwhile, among the hypoxia groups, although the control was significantly different from the sample groups, there was no statistically significant difference between the sample groups. When comparing the normoxia and hypoxia groups, there was a statistically significant difference between the control groups and sample groups treated with $1{\mu}g/mL$ of minocycline, yet no significant difference between the sample groups treated with $10{\mu}g/mL$ of minocycline. Conclusion : Minocycline was found to be neuroprotective in normoxia and hypoxia induced rat brain cortical cell cultures.

• Clinical and Experimental Pediatrics
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• v.49 no.3
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• pp.278-286
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• 2006

Purpose : Although suprapubic bladder aspiration(SBA) is the most reliable technique for identifying bacteriuria in young infant, no report has been published about the clinical manifestations of urinary tract infection(UTI) diagnosed by aspirated urine in Korea. This study was performed to examine clinical manifestations and related factors of UTI confirmed by a positive urine culture obtained by SBA in young infants. Methods : We reviewed medical records of 159 infants younger than six months of age, who had been admitted for UTI to Korea University Ansan Hospital from January 2002 to June 2005. Results : The male:female ratio was 5.1 : 1. The most common pathogen in urine culture was Escherichia coli(92.5 percent). Most of the gram-negative pathogens had high sensitivity to amikacin, or third generation cephalosporins and had low sensitivity to ampicillin, trimethoprim-sulfamethoxazole, or gentamicin. Hydronephrosis and vesicoureteral reflux(VUR) were present in 32.7 percent and 27.7 percent respectively. Renal cortical defects observed on DMSA scans were detected in 37.1 percent. Prolonged fever duration and hospital day, high erythrocyte sedimentation rate(ESR) and C-reactive protein(CRP) levels, hydronephrosis and VUR were related to the renal cortical defects(P<0.05). Rates of UTI-associated bacteremia and aseptic meningitis were 8.3 percent and 6.6 percent. Conclusion : Early differential diagnosis is very important in infants younger than 6 months of age with UTI because the clinical findings are non-specific and UTI-associated bacteremia and aseptic meningitis are concomitantly found. Because prolonged fever and higher ESR and CRP levels are risk factors of the renal cortical defects, radiologic evaluations and nephrologic follow-up were needed in identifying the predisposing congenital abnormalities and chronic renal scarring.