• The korean journal of orthodontics
• /
• v.27 no.2
• /
• pp.335-347
• /
• 1997

Vitamin D is known to exert its action by activating DNA and RBA within target cells to produce proteins and enzymes that can be used in bone resorption process. Particularly, the active form of vitmain D, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is considered to be one of the most potent stimulators of osteoclatic acitivity in vitro. The purpose of this study was to evaluate the effect of 1,25-Dihydroxyvitamin $D_3$ on the avtivity of periodotal ligament cells and, the experimental tooth movement. Human periodontal ligament cells were collected from the first premolar tooth extracted for the orthodontic treatment, and were incubated in the environment of $37^{\circ}C$, 5% $CO_2$ and 95% humidity. Microtitration(MIT) assay was done at 10, 25, 50 and 100ng/ml of 1,25-Dihydroxyvitamin $D_3$. 21 Sprague-Daft rats were divided into a control gmup(3), and experimental groups(18) where 100g of force from helical spring was applied across the maxillary incisors 1,25-Dihydroxyvitamin $D_3$ was injected into periodontal ligament at the mesial or distal surface of maxillary incisors so that we can compare the control side and the experimental side. Expreimental groups were sac rifled at 12, 24, 36, 48, 72hours and 7 days after force application, respectively. And the obtained tissues were evaluated histologically. The observed results were as follows. 1. The activity of periodontal ligament cells in l0ng/ml or 25ng/ml of 1,25-Dihydroxyvitamin $D_3$ 1,25-Dihydroxyvitamin $D_3$ was not significantly different to the control at the cultivation of 1, 2 and 3 days. 2. The activity of periodontal ligament cells was significantly increased at 3 days in 50 ng/ml of 1,25-Dihydroxyvitamin $D_3$ and 2, 3 days in 100g/ml of 1,25-Dihydroxyvitamin $D_3$. 3. Up to 7 days after force application, there was no difference in osteoblastic activity, tearing of periodontal ligament and proliferation of capillary at tension side between 1,25-Dihydroxyvitamin $D_3$ injection side and the control side. 4. The osteoclastic activity and the resorption of alveolar bone was greater in 1,25-Dihydroxyvitamin $D_3$ injection side than the control side at 36 hours after force application.

• Korean Journal of Animal Reproduction
• /
• v.10 no.2
• /
• pp.192-203
• /
• 1986

Two experiments in this study were designed to compare the potential for in vitro capacitation and in vitro fertilization of ejaculated sperm among individual rabbit bucks. In experiment 1, for in vitro capacitation, the ejaculated sperm were preincubated in DM for 12 hr or 18 hr after HIS treatment, then 12 hr -or 18 hr- preincubated sperm were incubated with superovulated rabbit ova in a 5% CO2 incubator for 36 hr at 38$^{\circ}C$, and a part of cleaved ova was transferred to the recipient does for implantation of embryo. In experiment 2, effect of lysolecithin addition to preincubation medium on induction of accelerated in vitro capacitation and in vitro fertilization of individual rabbit sperm was studied. Experiment 1; 1. Percent acrosome reaction of sperm, noted after staining, after 12 hr or 18 hr preincubation ranged from 52.5 to 76.0% and from 67.5 to 90.0%, respectively and sperm motility index of these sperm ranged from 20.0 to 47.5 for 12 hr-preincubated sperm and from 15.0 to 37.5 for 18 hr- preincubated sperm. There was no a certain relation between percent acrosome reaction and sperm motility index. 2. In vitro fertilization rate (cleavage rate) of in vitro capacitated sperm varied widely among individual bucks, ranging from 0 to 47.8% for 12 hr - preincubated sperm and from 0 to 60.9% for 18 hr -prein- cubated sperm. Cleavage rate of 18 hr - preincubated sperm was higher and faster than that of 12 hr - preincubated sperm. 3. Eight of 44 in vitro fertilized embryos transferred into 6 recipients were implanted in 4 recipients (66.7%) up to day 15 and implnatation rate was 18.2%. Experiment 2; 1. The percent acrosome reaction of sperm before and after 4 hr preincubation in DM without lysolecithin varied significantly among individual bucks, ranging from 0.4 to 18.4% and from 1.7 to 37.4%, respectively and percent acrosome reaction of sperm at 30 min after addition of 60${\mu}$g/ml lysolecithin also was significantly different among bucks, ranging from 19.2 to 67.1%. 2. Effect of accelerated acrosome reaction following lysolecithin addition was more considerable in the individuals showed less percent acrosome reaction before and after 4 hr preincubation. Percentage of motile sperm and motility score showed a trendency towards a decrease with increase of preincubation time and time after lysolecithin addition. 3. In vitro fertilization rate (cleavage rate) at 24 hr postinesmination with pooled sperm were treated to 60 $\mu\textrm{g}$/ml lysolecithin for 30 min after 4 hr preincubation was 24.6%, a higher rate than 13.2% for control. While 80 $\mu\textrm{g}$/ml lysolecithin-added sperm showed a lower cleavage than control and 60$\mu\textrm{g}$/ml-added sperm at both 24 hr and 48 hr postinsemination. These results from 2 experiments suggest that more useful preincubation time for the in vitro capacitation of ejaculated rabbit sperm is 18 hr in DM after HIS treatment, although there is wide variation in vitro capacitation and in vitro fertilization rate among individual bucks, and lysolecithin addition to at least 4 hr - preincubated sperm in DM can result in almost same in vitro fertilization rate as that of 18 hr - preincubated sperm in the experiment 1.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.26 no.2
• /
• pp.427-436
• /
• 1999

Preservation of the remaining periodontal ligament cells on an avulsed tooth is very important to the successful outcome of replantation. HBSS is recommended as the most suitable storage medium for the avulsed tooth that cannot be replanted immediately. But their availability near the site of an accident is doubtful. The purpose of this in vitro study was to compare periodontal ligament cells stored in different storage media obtained easily on the spot. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ culture medium containing 20% FBS, at $37^{\circ}C$ 100% humidity, in a 5% $CO_2$ incubator. Cells were cultured in 96 well culture plate, $5{\times}10^4$ cells per well with ${\alpha}-MEM$ and incubated for 24 hours. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS, pasteurized milk, sterilized saline, unstimulated saliva and bench-dried state at $25^{\circ}C$ room temperature for 30, 60, 90, 120, 180 minutes respectively. And then each group was measured using MTT assay. The results were as follows. 1. Between the group of each time, there was statistically significant difference. Periodontal ligament cells viability was highest in pasteurized milk and was reduced stepwisely in sterilized saline, unstimulated saliva and bench-dried state(p<0.05). 2. between the time of each group, there was statistically significant difference(p<0.05) but was no statistically significant difference at 90-120 minutes in pasteurized milk and at 60-90 minutes and 120-180 minutes in sterilized saline(p>0.05). In conclusion, HBSS as storage medium of an avulsed tooth is not practical on the spot. Insteadily pasteurized milk can be recommended to maintain the periodontal ligament cells viability.

• Journal of Embryo Transfer
• /
• v.16 no.3
• /
• pp.203-211
• /
• 2001

This study was carried out to determine sex of porcine embryos produced by in vitro fertilization. Porcine oocyte-cumulus complexes were cultured in BSA-free North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (10%), cystein (0.1 mg/ml) and hormonal supplement (10 IU eCG and 10 IU hCG per ml) for 20~22 hrs. They were then cultured in the same medium but without hormonal supplement for additional 20~22 hrs. After culture, cumulus cells were removed and oocytes were co-incubated for 6 hrs with four different concentrations (5$\times$10$^4$, 2.5$\times$ 10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) of porcine sperm. After fertilization, oocytes were transferred into NCSU 23 with 0.4% BSA medium. The cleavage and blastocyst formation rates were evaluated at 48 and 144 hrs, respectively. In this study, the polymerase chain reaction (PCR) was used to determine the sex of porcine embryos in the stage of blastocyst. The PCR was performed using a set of oligonucleotide primers (5‘-TCATGGACCAGGTAGGGAAT-3', 5’-GAAAGACACGTCCTTGGA GA-3') for 491 bp fragment of porcine male-specific DNA sequence. In the flour different sperm concentration (5$\times$10$^4$, 2.5$\times$10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) for fertilization condition, the cleavage rate was 55.95, 67.88, 60.18 and 47.60%, respectivety, and the development rate of blastocysts was 16.03, 20.40, 21.41 and 12.37%, respectively. At 5.0$\times$10$^4$and 2.5$\times$10$^{5}$ of sperm concentrations per ml cleavage rate and development rate of blastocyst were higher than those of 5.0$\times$10$^4$and l0$\times$10$^{5}$ of sperm concentration (P<0.01). The male of porcine embryos was detected at 491 bp by PCR, and 18 of the 31 porcine blastocysts were the male (58.1%) and the rest 13 were the female(41.9%).

• The Korean Journal of Nuclear Medicine Technology
• /
• v.13 no.3
• /
• pp.156-158
• /
• 2009

Purpose: Since radiopharmaceuticals are intended for human administration, it is imperative that we should undergo quality control very strictly. Now almost all the PET laboratories have adopted Bacterial Endotoxin Test as the stand quality control method to monitor whether pyrogen is free or not in the product vial containing crude solution. The aim of this study is to find out the reason why false positive result is observed when using commercially available test vial. Materials and Methods: For this experiment, we used commercially available single test kit (Associates of Cape Code. Inc. USA) and we made pH samples by mixing each buffer whose pH ranges are 1.0 to 12.0. Otherwise we made Ethanol samples diluted with distilled water. After making test samples, it added 0.2 mL to the test vial. Assay mixture in the test vial was incubated in a water bath (Chang Shin Co. KOR) for 60 min at $35{\pm}2^{\circ}C$. Results: After incubation period ($60{\pm}1^{\circ}C$), we inverted the test vial about $180^{\circ}$ To know what pH and how many percentage of Ethanol (Fisher Scientific Korea. Ltd) will affect the reaction. With pH buffer, false positive result was observed at pH 1.0 to 5.0 and 7.7 to 12.6 but at pH 5.2 to.7.5, the test results show negative. It's very strange that we couldn't observe negative test result with Tris buffer at pH 8.4, 8.6, 8.8, 9.0. in other case Ethanol, the test result was seen with 5 to 10% Ethanol. But to my surprise we could see very thick gel formation with 100% Ethanol. Conclusions: In this study, we could notice that pH which is too much acidic or alkalic or high concentrated Ethanol would affect Bacterial Endotoxin Test result. As you know, LAL test is sensitive and very reliable method. Therefore, we are needed to elicit the accurate test result as possible as we can.

• Journal of Mushroom
• /
• v.14 no.4
• /
• pp.220-224
• /
• 2016

Chronic inflammation, which results from continuous exposure to antigens, is one of major reasons for tissue damage and diseases such as rheumatoid arthritis and type 2 diabetes. In this study, we investigated the anti-inflammatory effects of extracts (hexane, $CHCl_3$, MeOH, $MeOH/H_2O$, and $H_2O$) from GW10-45, which is our new cultivar of an edible mushroom Pleurotus ferulae (ASI 2803 and ASI 2778), in RAW264.7 murine macrophages. None of the extracts showed cytotoxicity in RAW264.7 cells and the hexane, CHCl and H extracts reduced nitric oxide (NO) production, an important inflammatory marker, in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Particularly, the extract (CG45) inhibited NO production more than the other extracts did. To elucidate the effects of CG45 on molecular targets involved in pro-inflammatory responses, we performed western blot analysis. Expression of inducible nitric oxide (iNOS) significantly decreased in LPS and CG45 co-incubated cells compared to that in LPS only-treated cells. Additionally, another protein thatplays a critical role in inflammation, was down-regulated in cells treated with both LPS and CG45. In the nuclear factor $(NF)-{\kappa}B$ pathway, phosphorylation of $I{\kappa}B{\alpha}$ decreased in RAW264.7 cells treated with both LPS and CG45. Furthermore, CG45 inhibited the phosphorylation of $NF-{\kappa}B$ in LPS-stimulated RAW264.7 cells. Conclusively, CG45 could suppress pro-inflammatory responses in LPS-stimulated RAW264.7 cells by down-regulating not only the phosphorylation of $NF-{\kappa}B$ and $I{\kappa}B{\alpha}$ but also the expression of iNOS and COX-2 without any cytotoxicity.

• Asian-Australasian Journal of Animal Sciences
• /
• v.25 no.5
• /
• pp.690-700
• /
• 2012

The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen ($NH_3$-N) and $H_2S$ gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, $NH_3$-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as $NO_2$, $NO_3$ and $SO_4$ concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that $NH_3$-N, $H_2S$ and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds.

• The korean journal of orthodontics
• /
• v.28 no.4 s.69
• /
• pp.627-640
• /
• 1998

[$TGF-{\beta}$] is a polypeptide with multiple physiological functions in regulation of cell-to-cell interaction and in growth and development. The active form of vitmain $D_3$, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is one of the most potent stimulators of osteoclastic acitivity. The purpose of this study was to evaluate the effect of Vitamin $D_3$ and/or $TGF-{\beta}$ on the periodontal ligament(PDL) cells. Human PDL cells were prepared from the first premolars extracted for the orthodontic treatment and were incubated in the environment of , $37^{\circ},\;5\%\;CO_2\;and\;95\%$ humidity. 10, 50 or 100ng/m1 of $1,25-(OH)_2D_3$ and 0.1, 1, 5 or 10ng/ml of $TGF-{\beta}$ were administered to the culture wells, separately or in combination. And the viability of PDL cells was evaluated by MTT assay The obtained results were as follows. 1. The viability of PDL cells in 10ng/ml of vitamin $D_3$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 50ng/ml of Vitamin $D_3$ at 3-day and in 100ng/m1 of Vitamin $D_3$ at 2 and 3-day. 2. The viability of PDL cells in 0.1ng/ml of $TGF-{\beta}$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 1 and 5ng/ml of $TGF-{\beta}$ at 3-day of cultivation, and in 10ng/ml of $TGF-{\beta}$ at 2 and 3-day of cultivation. 3. In case of admixture of 1ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells was significantly increased in the admixture of 100ng/ml of vitamin $D_3$ at 3-day of cultivation 4. In case of admixture of 5ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells began to be increased from 2-day of cultivation in the admixture of 10 50 and 100ng/ml of vitamin $D_3$, but it was not maintained at 3-day in the admixture of 10ng/m of vitamin $D_3$. 5. In case of admixture of 10ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$ the viability of PDL cells was significantly increased in the admixture of 50ng/ml of vitamin $D_3$ at 2 and 3-day of cultivation, and in the admixture of 100ng/ml at 1, 2 and 3-day.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.42 no.2
• /
• pp.161-167
• /
• 2013

The aim of this study was to investigate the protective effects of methanolic extract from perilla (Perilla frutescens Britt var. japonica) leaves (PLME) on oxidative injury from hydrogen peroxide ($H_2O_2$) in human HaCaT keratinoctyes. Cells were co-incubated with various concentrations (0~200 ${\mu}g/mL$) of PLME for 24 hr, and then exposed to $H_2O_2$ (500 ${\mu}M$) for 4 hr. $H_2O_2$ significantly decreased cell viability (p<0.05). However, PLME provided protection from $H_2O_2$-induced HaCaT cell oxidation in a dose-dependent manner. To further investigate the protective effects of PLME on $H_2O_2$-induced oxidative stress in HaCaT cells, the cellular levels of lipid peroxidation, and antioxidant enzymes (including superoxide dismutase (SOD), glutathione peroxidase (GSH-px) and catalase (CAT)) were measured. PLME decreased cellular levels of lipid peroxidation, and also increased the activities of antioxidant enzymes. In addition, the antioxidant activities of PLME were also determined by DPPH and hydroxyl (${\cdot}OH$) radical scavenging assay, and major antioxidant compounds of PLME were measured by colorimetric methods. DPPH and ${\cdot}OH$ radical scavenging activities of PLME increased in a dose dependent manner and was similar to the DPPH scavenging activity of ascorbic acid at 50 ${\mu}g/mL$; however PLME activities were stronger than ascorbic acid (50 ${\mu}g/mL$) in the ${\cdot}OH$ scavenging assay. The amounts of antioxidant compounds, including total polyphenolics, total flavonoids, and total ascorbic acid from PLME were $52.2{\pm}1.1$ mg gallic acid (GAE)/g, $33.7{\pm}4.7$ mg rutin (RUE)/g, and $17.0{\pm}0.5$ mg ascorbic acid (AA)/g, respectively. These results suggest that PLME has a strong free radical-scavenging activity and a protective effect against $H_2O_2$-induced oxidative stress in the keratinocytes.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.23 no.2
• /
• pp.20-37
• /
• 2010

Purpose: The objective of this study was to observe the antibacterial effects of Sasangjasaebang (SSJSB) aqueous extracts, traditionally used for treating various gynecological diseases including vaginitis in Korea and their individual components - Cnidi Fructus(CF), Zanthoxyli Fructus(ZF) and Alumen(AL) against Gardnerella vaginalis ATCC14018, and combination effects of SSJSB extracts and Metronidazole were also monitored in this study. Methods: Antibacterial activities against Gardnerella vaginalis ATCC14018 of SSJSB (CF : ZF : AL = 10 : 3 : 3) aqueous extracts and their individual components were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at MIC and $MIC{\times}2$ levels. The combination effects of SSJSB aqueous extracts and Metronidazole were observed by Checkboard Microtiter Assay and effects on bacterial growth curve treated with SSJSB aqueous extracts MIC + Metronidazole MIC, 1/2MIC and 1/4MIC, respectively. In the present study, Gardnerella vaginalis ATCC14018 were incubated under $37^{\circ}C$, 10% CO2 and bacterial growth curves were calculated at 24, 48, 72, 96 and 120 hrs after incubations. Results: MIC of CF aqueous extracts, ZF aqueous extracts, AL powders and SSJSB aqueous extracts against Gardnerella vaginalis ATCC14018 were detected as $27.5\;{\pm}\;13.693$(12.5~50), $6.875\;{\pm}\;3.423$(3.125~12.5), $4.375\;{\pm}\;1.713$(3.125~6.25) and $2.188\;{\pm}\;0.856(1.563{\sim}3.125)mg/m{\ell}$, respectively. MIC of Metronidazole was detected as $7.6\;{\pm}\;5.367(2{\sim}16){\mu}g/m{\ell}$ at same conditions. In addition, Metronidazole, CF aqueous extracts, ZF aqueous extracts, AL powders, and SSJSB aqueous extracts showed marked dosage-dependent inhibition of bacterial growth, and more dramatical inhibitions were detected in Metronidazole 1/2 MIC + SSJSB aqueous extracts MIC treatment as compared with each of single Metronidazole MIC and SSJSB aqueous extracts MIC treatments, respectively. Especially, quite similar inhibitory effects on bacterial growth were detected in Metronidazole 1/4 MIC + SSJSB aqueous extracts MIC treatment as compared with single Metronidazole MIC treatment in the present study. FIC index were detected as $0.475\;{\pm}\;0.137$(0.375~0.625) at Checkboard Microtiter Assay. Conclusion: The results obtained in this study suggest that CF aqueous extracts, ZF aqueous extracts, AL powders, and SSJSB aqueous extracts showed antibacterial effects against Gardnerella vaginalis ATCC14018, and they also showed dosage-dependent inhibitory effects on the bacterial growth. More potent antibacterial effects were detected in SSJSB aqueous extract as compared with individual components, respectively. In addition, combination treatment of SSJSB aqueous extract and Metronidazole showed more potent inhibitory effects on the growth of Gardnerella vaginalis with FIC index $0.475\;{\pm}\;0.137$(0.375~0.625). According to these results, the combination of SSJSB aqueous extract and Metronidazole is synergistic, and it is expected that effective dosages of Metronidazole could be reduced to 1/4 levels in combination with SSJSB extracts. And it might be needed to make further studies to seek the herbs which have antibacterial effects on the Gardnerella vaginalis ATCC14018.