• Journal of Embryo Transfer
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• v.17 no.2
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• pp.145-151
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• 2002

The aim of these experiments was to investigate in vitro nuclear maturation of canine oocyte collected from various stages of estrus cycles, Ovaries were obtained from 1 to 4 year-old mongrel bitch and minced for oocyte collection in phosphate buffered saline with 100 iu penicillin-G $m\ell$／sup -1／, 50 $\mu\textrm{g}$ streptomycin sulphate $m\ell$／sup -1／ and 0.1％ (w／v) polyvinyl alcohol. Cumulus-oocyte-complexes (COCs) were washed in HEPES buffered tissue culture medium (TCM)199 and in vitro matured in TCM-199 culture medium supplemented with sodium pyruvate 0.028mg/$m\ell$, L-glutamine 0.146mg／$m\ell$, penicillin G 10,000 IU／$m\ell$, streptomycin 0.031mg／$m\ell$ and 10％ (v／v) fatal calf serum. COCs were in vitro matured for 48～72 hrs at 39$^{\circ}C$ in humidified 5％ $CO_2$ in air atmosphere. In vitro matured oocytes were remove the cumulus cells using 0.2％ (v／v) hyaluronidase. After denuding, oocyte were placed in acetic acid : methanol : chlorform solusion (3:6 : 1.5 v／v) for 30 sec and acetic acid: ethanol(1:3 v／v) for 48hrs fixation. Nuclear maturation was classified to GV, GVBD, MI, MII and degenerate oocyte under microscopy after 1％ aceto-orcein stain. In vitro maturation rates at 48hrs were not significantly difference among the oocytes collected from different stage of estrus at 15.9％, 16.3％, 23.7％ and 18.2％ for anestrus, proestrus, estrus and diestrus. However, the oocytes maturation(36.6％) of collected from estrus ovaries were significantly different from oocytes derived from proesturs, diestrus and anestrus ovaries(30.8％, 17.5％ and 22.1％; p＜0.05). The overall in vitro maturation rates were significantly higher (p＜0.05) in 72hrs culture than 48hrs culture system. In summary, there was a tendency for higher in vitro maturation rates with the oocyte collected from estrus ovary than other stages of estrus. Also, for nuclear maturation, in vitro culture of oocyte for 72hrs was better than 48hrs culture.

• Tuberculosis and Respiratory Diseases
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• v.57 no.5
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• pp.425-433
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• 2004

Background : Induction of oral tolerance (OT) has been known to prevent allergic inflammation in acute asthma model within 4 weeks. However it is remained whether induction of OT may effectively prevent allergic inflammation in chronic asthma model over 4 weeks. We observed the effect of induction of OT on allergic inflammation and airway remodeling in chronic asthma model up to 8 weeks. Methods : 5-week-old female BALB/c mice divided into 4 groups-control group, asthma group, low dose OT group, and high dose OT group. To induce oral tolerance mice were fed ovalbumin (OVA) before sensitization with OVA and aluminum hydroxide-1 mg for 6 consecutive days in the low dose OT group and 25 mg once in the high dose OT group. Mice in the asthma group were fed phosphate buffered saline instead of OVA. After sensitization followed by repeated challenge with aerosolized 1% OVA during 6 weeks, enhanced pause (Penh), inflammatory cells, IL-13, and IFN-${\gamma}$ levels in bronchoalveolar lavage (BAL) fluids as well as OVA-specific IgE, IgG1, and IgG2a levels in serum were measured. In addition the degree of goblet cell hyperplasia and peribronchial fibrosis were observed from lung tissues by PAS and Masson's trichrome stain. Results : Both OT groups showed a significant decrease in Penh, inflammatory cells, IL-13, and IFN-${\gamma}$ levels in BAL fluids as well as OVA-specific IgE, IgG1, and IgG2a levels in serum compared with the asthma group (P<0.05). In addition, the degree of goblet cell hyperplasia and peribronchial fibrosis were significantly attenuated in both OT groups compared with the asthma group (P<0.01). Conclusion : These results suggest that induction of OT may effectively prevent allergic inflammation as well as airway remodeling even in chronic asthma model up to 8 weeks.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.355-360
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• 2010

Vibrio parahaemolyticus (V. parahaemolyticus), which is commonly found in raw seafood, causes gastroenteritis in humans. Rapid and effective methods have been developed as culture methods require up to 5-7 days. In this study, real-time PCR was compared with the standard culture method for detecting V. parahaemolyticus in seafood and radish sprout samples. Five hundred grams of the samples were artificially contaminated with V. parahaemolyticus then divided into 20 samples. The samples were incubated in alkaline peptone water and then streaked onto thiosulfate-citrate-bile saltssucrose agar. Biochemical tests for suspicious colonies were performed using the API 20NE strip. In parallel, real-time PCR was performed targeting the toxR gene using the enrichment broth. The real-time PCR was sensitive in discriminating V. parahaemolyticus from other foodborne pathogens. The detection limit of the real-time PCR was $10^3\;CFU/mL$ in phosphate-buffered saline. Although the real-time PCR detected more positive samples (76 out of 180, 42%) than the culture method (66 out of 180, 37%), there was no significant statistical difference (p>0.05) between the two methods. In conclusion, real-time PCR assays could be an alternative to the standard culture method for detecting V. parahaemolyticus in seafood and radish sprouts, which has many advantages in terms of detection time, labor, and sensitivity.

• Journal of Animal Science and Technology
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• v.48 no.6
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• pp.797-804
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• 2006

The present study was undertaken to investigate the post-thawed survivability of bovine embryo depending on different dose of ethylene glycol and sucrose. Ovaries were collected at local slaughterhouse and the cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured at 39°C in an atmosphere of 5% CO2 incubator. For conventional slow-freezing, d 7 or 8 expanded blastocysts were collected. Embryos were equilibrated in 1.5 M and 1.8 M ethylene glycol(EG) with 0.1 M and 0.3 M sucrose in Dulbecco's phosphate-buffered saline(D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25ml-straw and placed directly into cooling chamber of programmable freezer precooled to 󰠏7°C, after 2 min, the straw was seeded, maintained at 󰠏7°C for 8 min, and then cooled to 󰠏35°C at 0.3°C/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 sec and exposed to 37°C water for 20 sec. Straws were then removed from 37°C water. Rates of blastocyst survive and hatching were evaluated at 24 to 72 h post-warming. No difference of the survivability was shown between 1.5 M and 1.8 M EG (71 and 70%, respectively). Addition of 0.1 M sucrose to 1.5 M and 1.8 M ethylene glycol in the freezing solution did not differ significantly embryo survival (74 and 77%, respectively), whereas survival rates was higher(89%) in freezing solution contained 0.3M sucrose to 1.8M EG compared with 0.3M sucrose to 1.5M EG group(71%). However, there was no difference in the overall total cell number between the two groups (122±1.8 vs 131±1.4, respectively). In conclusion, the results suggest that 0.3 M sucrose in 1.8 M EG may be optimal condition for freezing and thawing methods with in vitro produced embryos and may be applied to on-farm conditions for embryo transfer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.9
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• pp.1126-1135
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• 2008

This study was carried out to investigate the anti-oxidative and anti-inflammatory actions of genistein in BALB/c mice injected with lopopolysaccharide (LPS), called endotoxin. Mice (10 weeks of age) weighing approximately 20 g were divided into 4 groups. Endotoxin shock was induced by intraperitoneal injection of LPS (100 mg/kg BW). LPS and genistein+LPS groups were injected with LPS 30 min after phosphate buffered saline (PBS) solution and genistein (200 mg/kg BW) injections, respectively. Genistein group was injected with genistein, followed by PBS, while PBS group received two injections of PBS. Superoxide anion generation of peritoneal macrophage cells was significantly (p<0.05) lower in the genistein+LPS group than in the LPS injection group at 8 h after intraperitoneal injection, while SOD activity was significantly higher in genistien+LPS group than LPS group. Tumor necrosis factor-$\alpha$ levels of plasma were significant lower (p<0.05) in the genistein+LPS injection group than LPS group at 8 h after intraperitoneal injection. Plasma TBARS was lower in genistein+LPS group than LPS group, while hepatic TBARS were not different among groups. Hepatic glutathione concentrations and antioxidant enzyme activities were ignificantly higher in the genistein+LPS group than in the LPS group at 1 h and 8 h after intraperitoneal injection. Nuclear factor-kappa B (NF-${\kappa}B$) transactivation was significantly (p<0.05) inhibited in LPS group. These results demonstrate genistein may ameliorate inflammatory diseases through inhibition of NF-${\kappa}B$ transactivation and oxidative stress, which may be mediated partially by anti-oxidative effect of genistein.

• Journal of Korean Ophthalmic Optics Society
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• v.20 no.4
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• pp.463-469
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• 2015

Purpose: A hydrogel including alginate and $CaCl_2$ extracted from seaweed was manufactured, and their physical properties were investigated. Also, its applicability as contact lenses was examined. Methods: A film-type sample used in this experiment was manufactured using 2-hydroxyethyl methacrylate (HEMA), which is the raw material of hydrogel contact lenses; azobisiobutyonitile (AIBN), which is an initiator and ethylenglycoldimethacrylate (EGDMA), which is a cross-linking agent. It was hydrated in a PBS solution for 24 hours, and an interpenetrating polymer network (IPN) was formed in 1% and 2% alginate and 1%, 3%, and 5% $CaCl_2$ solutions for 24 hours, respectively. Results: The measurement of the physical properties of the film after the IPN showed that the moisture content was 30.89~36.89%, the refractive index was 1.431~1.441, the contact angle was $62.98{\sim}80.45^{\circ}$, and the tensile strength was 2.378~4.215 ($gf/mm^2$). Also, the physical properties hardly changed as the content of alginate increased, and the moisture content decreased as the content of $CaCl_2$ increased. As a result of the IPN, the moisture content and contact angle decreased compared to those of basic HEMA, but the tensile strength increased. The tensile strength of the second IPN was higher than that of the first IPN. In the case of $CaCl_2$, for the sample polymerized for 24 hours and the second IPN sample with 2% alginate, the contact angle decreased as the content of $CaCl_2$ increased. Conclusions: In this study, the tensile strength increased as the content of $CaCl_2$ increased, and the wettability increased as a result of IPN of alginate and $CaCl_2$. The hydrogel containing the alginate and $CaCl_2$ was confirmed possible utilization as contact lens material.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.6
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• pp.783-788
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• 2016

Excessive intake of trans fats is a risk factor for cardiovascular diseases. Previously, we showed that green tea extract lowers intestinal absorption of cholesterol and elaidic acid in rats. This study was conducted to investigate a possible role for sorghum extract on lymphatic absorption of trans fat and cholesterol in rats. Adult male rats with lymph cannulae were infused at a rate of 3.0 mL/h for 8 h via a duodenal catheter with a lipid emulsion containing $146.4{\mu}mol$ of trielaidin, $36.8{\mu}mol$ of trilinoelaidin, $452.0{\mu}mol$ of triolein, $1.0{\mu}Ci$ cholesterol labeled with $^{14}C$ ($^{14}C-cholesterol$), $20.7{\mu}mol$ of cholesterol, and $396.0{\mu}mol$ of Na-taurocholate without or with 100.0 mg of sorghum extract in phosphate-buffered saline buffer (pH 6.4). Lymph was collected hourly for 8 h. No significant difference was noted in lymph flow. However, the lymphatic absorption of elaidic acid and linoelaidic acid for 8 h was significantly lower in rats infused with sorghum extract than in those infused with no sorghum extract. Further, lymphatic absorption of $^{14}C-cholesterol$ was reduced by sorghum extract, which was observed previously. These data indicate that sorghum extract has an inhibitory effect on the intestinal absorption of trans fat and cholesterol. The mechanism(s) by which sorghum extract lowers intestinal absorption of trans fat warrants further study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.11
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• pp.1415-1421
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• 2008

Preventive effects of 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid (HDMPPA), an active compound in Korean cabbage kimchi with anti-atherogenic effects, on the accumulation of lipids in the vital organs of $apoE^{(-/-)}$ mice fed atherogenic diet (AD) were studied. Each group of 10 mice was fed AD for 8 weeks with intraperitoneal injection of either HDMPPA (1 mg HDMPPA/100 g BW/day) or phosphate buffered saline as a vehicle. The organs used for this study were liver, kidney, spleen, lung, testis, and brain. Total cholesterol (TC) concentration of lung was the highest followed by spleen and brain. TC level for the liver was the lowest. In contrast to the results of TC, triglyceride (TG) concentration in the liver was the highest followed by kidney and testis. $ApoE^{(-/-)}$ mice did not have any problem uptaking chylomicron remnant by the liver which carries an extra TG after delivering it to the adipose tissue. HDMPPA retarded TC and TG accumulations in the vital organs. Thiobarbituric acid reactive substances (TBARS) in the brain and spleen were the highest and that in the testis were the lowest. Poly-unsaturated fatty acids in the brain and activated peroxisome in the spleen might be responsible for high TBARS levels in these organs. The greatest antioxidant effect of HDMPPA against lipid peroxidation was observed in the spleen, showing 21.47% decrease. The most noticeable effect of HDMPPA was observed in glutathione (GSH) level. GSH levels of six organs in the HDMPPA group were significantly higher than those of the control group. GSH-peroxidase activity was negatively related to GSH level of the organs except liver and spleen. In conclusion, HDMPPA from Korean cabbage kimchi inhibits the lipid accumulation as well as increases the antioxidant status in the vital organs of $apoE^{(-/-)}$ mice fed an atherogenic diet.

• Tuberculosis and Respiratory Diseases
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• v.41 no.3
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• pp.270-276
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• 1994

Background: The measurement of nonspecific bronchial hyperreactivity is valuable for diagnosis and management of bronchial asthma. Methacholine or histamine is used for the pharmacologic provocation test. Usually a methacholine bronchial provocation test is performed by a dosing technique with counted number of breaths. A dosimeter is indispensable in the dosing technique. Recently a timed tidal breathing technique which dose not need an expensive dosimeter was introduced. We measured the degree of nonspecific bronchial hyperreactivity to histamine using a simple timed tidal breathing technique. Method: Forty two healthy volunteers, 12 patients with bronchial asthma(BA), 10 patients with rhinitis(RH) and 10 patients with upper respiratory infection(URI) participated in the study. The subject's nose was clipped and inhalation continued during tidal breathing for 2 minutes via a face mask. $FEV_1$ was measured at 30 seconds, 90 seconds after inhalation and inhalation of next solution was continued until there was a fall in $FEV_1$ of 20%. Histamine PC20 was defined as the concentration at 20% fall of $FEV_1$ and it was obtained from the log dose-response curve by linear interpolation. Results: Inhalation of serial dilution of histamine could be performed in all patients without significant side effects. The geometric mean${\pm}$standard deviation of histamine PC20 in healthy volunteers is $8.27{\pm}2.22mg/ml$, BA group $0.33{\pm}3.02mg/ml$, RH group $0.85{\pm}3.24mg/ml$, and URI group $1.47{\pm}1.98mg/ml$. Conclusion: Histamine bronchial provocation test using timed tidal breath method is a simple and suitable tool for management of patients with bronchial hyperreactivity.

• Applied Chemistry for Engineering
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• v.34 no.3
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• pp.258-263
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• 2023

In this study, conductive polymers and the enzyme tyrosinase (Tyr) were deposited on the surface of a screen printed carbon electrode (SPCE), which can be fabricated as a disposable sensor chip, and applied to the detection of bisphenol F (BPF), an endocrine disruptor with proven links to male diseases and thyroid disorders, using electrochemical methods. On the surface of the SPCE working electrode, which was negatively charged by oxygen plasma treatment, a positively charged conductive polymer, poly(diallyldimethyl ammonium chloride) (PDDA), a negatively charged polymer compound, poly(sodium 4-styrenesulfonate) (PSS), and another layer of PDDA were layered by electrostatic attraction in the order of PDDA, PSS, and finally PDDA. Then, a layer of Tyr, which was negatively charged due to pH adjustment to 7.0, was added to create a PDDA-PSS-PDDA-Tyr sensor for BPF. When the electrode sensor is exposed to a BPF solution, which is the substrate and target analyte, 4,4'-methylenebis(cyclohexa-3,5-diene-1,2-dione) is generated by an oxidation reaction with the Tyr enzyme on the electrode surface. The reduction process of the product at 0.1 V (vs. Ag/AgCl) generating 4,4'-methylenebis(benzene-1,2-diol) was measured using cyclic and differential pulse voltammetries, resulting in a change in the peak current with respect to the concentration of BPF. In addition, we compared the detection performance of BPF using an ionic liquid electrolyte as an alternative to phosphate-buffered saline, which has been used in many previous sensing studies. Furthermore, the selectivity of bisphenol S, which acts as an interfering substance with a similar structure to BPF, was investigated. Finally, we demonstrated the practical applicability of the sensor by applying it to analyze the concentration of BPF in real samples prepared in the laboratory.