• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.235-243
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• 2000

Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1) enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at $37^{\circ}C$ and 5% $CO_2$ incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 lU/ml and DNAse 20 lU/ml. After 20 min., follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when ${\rho}$ value was less than 0.05. Results: In handling time, mincing or scraping method ($28{\pm}3.42$ min or $16{\pm}1.58$ min) were significantly (p<0.00001) shorter than enzymatical method ($72{\pm}1.69$ min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method ($49.8{\pm}3.91$) than in mincing or scraping method ($25.3{\pm}2.33$ or $20.5{\pm}1.75$). Isolated follicles in ${\leq}$90${\mu}m$ were significantly (p<0.005) higher in enzymatical method ($15{\pm}1.71$) than in mincing or scraping method ($7.8{\pm}0.98$ or $8.1{\pm}1.31$). In 91~130 ${\mu}m$, isolated follicles were significantly (p<0.0005) higher in enzymatical method ($33{\pm}3.27$) than in mincing or scraping method ($16.3{\pm}1.82$ or $10.7{\pm}1.38$). In ${\geq}$ 131 ${\mu}m$, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ${\leq}$ 90 ${\mu}m$ was highest in scraping method (39.6% vs. enzymatical method: 30.1%, p<0.05; mincing method: 30.9%, p=0.11719, NS). Rate of follicles in $91{\sim}130$ ${\mu}m$ was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ${\geq}$131 ${\mu}m$ was highest in scraping method (8.3% vs. enzymatical or scraping method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handling time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91 ~ 130 ${\mu}m$ was highest in all methods.

• Journal of Life Science
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• v.22 no.12
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• pp.1621-1627
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• 2012

Cell motility plays an essential role in many physiological responses, such as development, immune reaction, and angiogenesis. In the present study, we showed that lysophosphatidic acid (LPA) modulates cancer cell migration by regulation of generation of reactive oxygen species (ROS). Stimulation of SKOV-3 ovarian cancer cells with LPA strongly promoted migration. but this migration was completely blocked by pharmacological inhibition of phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. Inhibition of the ERK pathway had no effect on migration. Stimulation of SKOV-3 ovarian cancer cells with LPA significantly induced the generation of ROS in a time-dependent manner. LPA-induced generation of ROS was significantly blocked by pharmacological inhibition of PI3K or Akt, but inhibition of the ERK signaling pathway had little effect. LPA-induced generation of ROS was blocked by pretreatment of SKOV-3 ovarian cancer cells with an NADPH oxidase inhibitor, whereas inhibition of xanthine oxidase, cyclooxygenase, or mitochondrial respiratory chain complex I had no effect. Scavenging of ROS by N-acetylcysteine completely blocked LPA-induced migration of SKOV-3 ovarian cancer cells. Inhibition of NADPH oxidase blocked LPA-induced migration whereas inhibition of xanthine oxidase, cyclooxygenase, or mitochondrial respiratory chain complex I did not affect LPA-induced migration of SKOV-3 ovarian cancer cells. Given these results, we suggest that LPA induces ROS generation through the PI3K/Akt/NADPH oxidase signaling axis, thereby regulating cancer cell migration.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.69-73
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• 2003

This study was undertaken to investigate the effect of three different porcine IVM media, TCM-199, mSOF and NCSU-23 on early development of porcine spontaneous parthenogenotes. Spontaneous parthenogenotes were induced by a prolonged culture of porcine oocytes in each medium. In Experiment 1, oocytes derived from gilts were matured in three IVM media and maturation of oocytes was evaluated by the status of chromatin configuration. Oocytes matured in mSOF, NCSU-23 and TCM-199 showed no significant difference (P>0.05) in maturation. Maturation rates at 48h after IVM were 83.1$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (mSOF), 78.0$\pm$3% (NCSU-23) and 83.5$\pm$2% (TCM-199). In Experiment 2, pronucleus formation and development to 6~8 cell stage of pig oocytes activated spontaneously. Pronucleus formation, cleavage rate and development to 6~8 cell embryos of porcine spontaneous parthenogenotes were assessed at 55~58 h, 96 h and 168h after IVM, respectively. Pronucleus formation (5.4$\pm$2% and 3.7 $\pm$ 1% vs 1.7 $\pm$ 3%) and development to the 6$\pm$8 cell (3.2$\pm$3% and 4.0$\pm$1% vs 1.4$\pm$3%) was significantly (P<0.05) higher in mSOF or NCSU-23 than TCM-199. In conclusion, the present study showed that oocytes matured in mSOF and NCSU-23 were spontaneously activated with higher frequency.

• Journal of agriculture & life science
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• v.45 no.2
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• pp.43-50
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• 2011

Hibiscus cannabinus L. is a plant in the Malvaceae family, that was seeded at June 1st in 2010 and harvested at November 18th. The present study was designated to investigate the safety for host cells, antibacterial effects of Hibiscus cannabinus L. of flower (HCME-F) or leaf (HMEF-L) methanol extract for typical Gram's positive bacteria (St. aureus and Str. epidermidis) or Gram's negative bacteria (S. typhimurium and E. coli). In treatment of different concentrations of HCME-F or HMEF-L (1, 50 and $100{\mu}g/ml$), cytotoxic effects were not shown to RAW 264.7 cells until 24 h incubation. In determination of antibacterial activity of HCME-F or HMEF-L, the antibacterial activities for St. aureus and Str. epidermidis were markedly increased compared to that of untreated control group, but antibacterial activity of HCME-F or HMEF-L for S. typhimurium and E. coli were not changed. Taken together, we demonstrated that methanol extract of HCME-F or HMEF-L showed the safety for RAW 264.7 cells and antibacterial activities for Gram's positive pathogenic bacteria St. aureus and Str. epidermidis. These findings suggest that a methanol extract of Kenaf flower or leaf may be useful alternatives of conventional chemotherapies for dermatitis and mastitis causing Gram's positive pathogens such as Stapylococcus spp. and Streptococcus spp. in domestic animals and humans.

• Korean Journal of Food Science and Technology
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• v.51 no.5
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• pp.480-485
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• 2019

The present study was conducted to compare antioxidant capacities of protein hydrolysates from four different edible insects (Protaetia brevitarsis larvae, Allomyrina dichotoma larvae, Gryllus bimaculatus imago, and Tenebrio molitor larvae) which have recently been registered as food varieties in Korea. Protein hydrolysates were prepared from each insect using enzymatic hydrolysis using alcalase, and were then separated into a fraction containing ${\leq}3kDa$. According to $RC_{50}$ values and trolox equivalent antioxidant capacity results obtained from five different antioxidant analyses, the Gryllus bimaculatus (GB) hydrolysate showed relatively high levels of antioxidant capacity and, in particular, the GB hydrolysate showed considerably strong antioxidant activities in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and in ferric reducing antioxidant power (FRAP) assays. The GB hydrolysate also showed the strongest inhibitory effect on peroxidation of linoleic acid, and its rate of inhibition at $100{\mu}g/mL$ on day 3 of treatment was 60.26%. These results suggest that protein hydrolysates from edible insects including GB represent potential sources of natural antioxidants.

• Korean journal of applied entomology
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• v.61 no.1
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• pp.155-163
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• 2022

The application of fungicides is indispensable to global food security, and their use has increased in recent times. Fungicides, directly or indirectly, have impacted insects, leading to genetic and molecular-level changes. Various detoxification mechanisms allow insects to eliminate reactive oxygen species (ROS) toxicity induced by agrochemicals including fungicides. In the present study, we analyzed the mRNA expression levels of detoxifying enzymes in Tenebrio molitor larvae following exposure to non-lethal doses (0.2, 2, and 20 ㎍/µL) of a fungicide captan. Transcripts of peroxidases (POXs), catalases (CATs), superoxide dismutases (SODs), and glutathione-s-transferases (GSTs) were screened from the T. molitor transcriptome database. RT-qPCR analysis showed that TmPOX5 mRNA increased significantly 24 h post-captan exposure. A similar increase was noticed for TmSOD4 mRNA 3 h post-captan exposure. Moreover, the expression of TmCAT2 mRNA increased significantly 24 h post-treatment with 2 ㎍/µL captan. TmGST1 and TmGST3 mRNA expression also increased noticeably after captan exposure. Taken together, these results suggest that TmPOX5 and TmSOD4 mRNA can be used as biomarkers or xenobiotics sensors for captan exposure in T. molitor, while other detoxifying enzymes showed differential expression.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.457-463
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• 2023

To investigate residual pesticide levels in agricultural products contained in Meal-kits, 27 Meal-kit products were collected from marts, Meal-kit shops, and online stores in Incheon City, South Korea. Seventy-six vegetable and thirty-seven mushroom products were analyzed for residual levels of 339 pesticides. Residual pesticides were detected in 23 out of 76 vegetables and were not present in the 37 mushroom products. The residual pesticide detection rate was 20.4% (23/113 cases). The pesticides famoxadone 0.034 mg/kg (standard: 0.01 mg/kg or less, PLS) and fenpyroximate 0.302 mg/kg (standard: 0.01 mg/kg or less, PLS) exceeded their maximum residue levels (MRL). This survey revealed that various types of pesticides remain in agricultural products in Meal-kits. Due to the nature of Meal-kit products, there is no separate standard for residual pesticides in agricultural products. Therefore, continuous monitoring of residual pesticides is necessary.

• Journal of the Korean Academy of Child and Adolescent Psychiatry
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• v.9 no.2
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• pp.174-179
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• 1998

Objective：Present study investigated the relationship between mothers' self-perception and perception of children's problems. Method：Subjects were 63 children, mothers, who visited child & adolescent psychiatric clinic. Their mothers completed Minnesota Mulpiphasic Personality Inventory(MMPI) & Korean Personality Inventory for Children(KPI-C). Correlation between validity scales of mothers' MMPI and validity scales of KPI-C was analysed, and then we divided subjects into two group(High & Low Group) according to the MMPI validity scales, and compared KPI-C's validity scales by student t-test. Result：There were significant positive correlation(r=.30) between K scale of MMPI and L scale of KPI-C, significant negative correlation(r=-.32) F scale of MMPI and L scale of KPI-C. The Means of T scores did show significant difference according to MMPI F, K scales in KPI-C L scale, to MMPI L scale in KPI-C F scale. Conclusion：These results indicated that defensive mothers tend to perceive her child's problems more defensively. However mothers who exaggerate their problem more severely, do not exaggerate their children's problem. Therefore, these result suggested that we should be careful not to underestimate children's problem whose mother elevated in K scale of MMPI.

• Journal of Animal Science and Technology
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• v.54 no.2
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• pp.141-147
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• 2012

The present survey was conducted to provide basic information on Hanwoo calf management. Eight hundred and sixty-two Hanwoo breeding farms from all nine provinces were surveyed via personal interviews. The percentages of farms categorized by herd size were 30.5%, 32.8%, 26.0% and 10.7% for <50 heads, 51-100 heads, 101-200 heads, and >200 heads, respectively. More than 50% of farms offered calf starter at 6-10 days of age, showing that calf starter was offered relatively at an earlier age. Calf starter was replaced every three days by 30.1% of farms. The percentages of farms replacing starter weekly (19.2%) were even higher than those of replacing starter daily (18.8%), suggesting that the frequency of replacing starter needs to be increased to maintain starter freshness and to increase starter intake. About one-third of farms offered forage at 6-10 days of age and 21% of farms offered even at 1-5 days of age although it has been well known that forage does not contain either nutrient density or nutrient profile necessary to stimulate rumen papillae development, especially before weaning. Furthermore, about half of farms used rice straw with calf starter. Water was offered relatively at an earlier age (1-5 days of age) by 55% of farms. Deciding when to wean calves should be based on starter intake rather than age but less than 50% of farms decided weaning age by starter intake. In conclusions, to reduce weaning age of Hanwoo calves by rapid rumen papillae development it is necessary to provide fresh starter and water by increasing frequency of starter replacing and water trough cleaning and not to feed forage before weaning.

• Journal of Chest Surgery
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• v.43 no.6
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• pp.576-587
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• 2010

Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.