• Journal of the Society of Cosmetic Scientists of Korea
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• v.39 no.1
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• pp.19-24
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• 2013

To investigate the efficacy of enzymatic extract of Ecklonia cava and its polyphenol extract (AG-DK) as cosmetic ingredients, the anti-oxidative effect, anti-glycation effect, anti-melanogenic effect, and anti-inflammatory effect of the extracts were evaluated in vitro. The enzymatic extract of E. cava ($SC_{50}$ 42.9 ppm) and AG-DK ($SC_{50}$ 6.4 ppm) showed a strong DPPH free radical scavenging activity. The anti-glycation ability of the enzymatic extract of E. cava and AG-DK was tested using bovine serum albumin (BSA), which inhibited the formation of advanced glycation end-products (AGEs) in the BSA/glucose system. The enzymatic extract of E. cava ($IC_{50}$ 97.2 ppm) and AG-DK ($IC_{50}$ 7 ppm) had inhibitory effects on tyrosinase activity. Moreover, the enzymatic extract of E. cava and AG-DK had an anti-inflammatory effect through the inhibition of nitricoxide (NO) and prostaglandin E2 ($PGE_2$). These findings suggest that the enzymatic extract of E. cava and AG-DK can be applied to skin-care products as cosmetic ingredients.

• Food Science and Preservation
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• v.22 no.1
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• pp.19-26
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• 2015

Indole 3-carbinol (I3C), important component of cruciferous vegetables and its major acid-catalyzed metabolite, 3,3'-diindolylmethane (DIM) have been suggested to have an inhibitory effect on the tumor growth and metastasis. This study investigated the effect of DIM on the adhesion, migration and invasion of highly invasive PC3 and DU145 human prostate cancer cell lines. Cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 3.0 g/L glucose, 3.7 g/L sodium bicarbonate and 10% fetal bovine and were incubated in a humidified incubator at $37^{\circ}C$ and 5% $CO_2$. DIM reduced the adhesion of PC3 and DU145 cells in a dose dependent manner. The pretreatment of PC3 cells with DIM reduced the adhesion dose dependantly, but inhibition was less effective than the treatment with DIM during the adhesion assay. The migration and invasion of PC3 and DU145 cells were reduced by DIM dose dependantly, and the inhibition of DIM was less effective in the DU145 cells than in the PC3 cells. The pretreatment of PC3 cells with DIM for 24 hr before the assay reduced invasion of PC3 cells by 37%. These results suggest that DIM inhibits adhesion, migration and invasion of the PC3 and DU145 cells and may be an effective antimetastatic therapy in addition to traditional chemotherapy.

• Archives of Pharmacal Research
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• v.19 no.2
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• pp.116-121
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• 1996

Antisense oligonucleotides seem to provide a promising new tool for the therapy. Choi et al. (1995) reported antisense phosphorothioate oligonucleotides (PS-ODN, 25 mer) complementary to TGF-.betha. mRNA designed for scar formation inhibitor to eliminate scars, which was caused by undesired collagen deposition due to overexpression of TGF-.betha., in wounded skin. PS-ODN were evaluated in vitro for skin penetration using normal and tape-stripped damaged rat skin. The in vitro skin transports were carried out with partially modified PS-ODN (6S) and fully modified PS-ODN (25S). The cumulative amount of PS-ODN (6S) penetrated through normal rat skin was $0.234{\pm}0.041{\mu}g/cm^2$ and that of tape-stripped damaged rat skin was $1.077{\pm}0.301{\mu}g/cm^2$ over 8 hrs. PS-ODN (25S) can not be found in receptor medium through normal skin due to high molecular weight (Mol.Wt.=8,000) and polyanionic charge. However, the cumulative amount of PS-ODN (25S) penetrated across damaged rat skin in PBS was $0.340{\pm}0.296{\mu}g/cm^2$ over 8 hrs. The absense of dermis raised the cumulative amount of PS-ODN (6S) penetrated through rat skin. And the fluxes of PS-ODN (6S) and PSODN (25S) at 8hrs across damaged rat skin were $134.63{\pm}37.67{\mu}g/cm^2$ h, and $42.50{\pm}36.95ng/cm^2$ h, respectively. While PS-ODN (25S) was stable in 10% heat inactivated fetal bovine serum (FBS) during 24 hrs, PS-ODN (6S) was less stable than PS-ODN (25S), but was markedly stable than unmodified phosphodiester. It is suggested that the cumulative amount of PS-ODN (6S) penetrated through damaged rat skin is larger than that of PS-ODN (25S) since the former is easier to degrade by nuclease than the latter and then is apt to penetrate into skin. Thus, PS-ODN represents a logical candidate for further evaluation due to the potential for delivery into the wounded skin.

• Korean Journal of Food Science and Technology
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• v.20 no.2
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• pp.170-175
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• 1988

This experiment was carried out to study the properties of casein micelles obtained from colostral skim milk. As lactation was progressed from parturition until 240h after calving, the content of total protein decreased while the proportion of casein to whey protein increased. Fractionaltion according to the site of casein micelle was done by ultracentrifugation at 100,000 x g for 10 minutes(pellet 1), 30 minutes(pellet 2) and 60 mintes(pellet 3) and the serum casein was prepared by acid precipitation of final supernatant at pH 4.6. During the lactation period, the relative amount of pellet 1(large size) decreased, that of pellet 2(middle size) maintained nearly constant level except for pllet from parturition, that of pellet 3(small size) increased, and the serum casein showed almost constant level. The relative amounts of ${\alpha}_{s1}-casein\;and\;{\alpha}_{s2}-casein\;and\;{\beta}-casein-5P$ in the pellets decreased and that of x-casein increased markedly with decreasing micelle size, but the relative amounts of ${\beta}-casein-1P$(f 29-209), (f 106-209) and (f 108-209) showed little change. The composition of the serum casein was different from that of the skim milk casein.

• The Journal of Korean Academy of Prosthodontics
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• v.44 no.1
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• pp.112-123
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• 2006

Purpose: The biocompatibility and bio-adhesive property of a dental implant abutment are important for proper soft tissue healing and maintenance of osseointegration of implant. However, studies of soft tissue healing and mucosal attachment of various materials of implant abutment other than titanium are still needed. In this study, cell attachment, proliferation, cytotoxicity of human gingival fibroblast for ceramic, gold alloy, Ni-Cr alloy and, commercially available pure titanium as a control were evaluated, using MTS and scanning electron microscopy. Materials and Methods: Specimen was designed to disc, 4mm diameter and 1mm thickness, made of ceramic, gold alloy, Ni-Cr alloy and commercially available pure titanium. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. Cells were inoculated in the multiwell plates placed the specimen disc. Cell Titer 96 AQucous One Solution Cell Proliferation Assay were done after 1hour 3hours, 24hours, 3days, 5days of incubation. The discs were processed for scanning electron micrography to evaluate cell attachment and morphologic change. Results: The results were obtained as fellows. 1. The ceramic showed high cell attachment and proliferation and low cytotoxicity, which is as much bioadhesive and biocompatible as titanium. 2. The gold alloy represented limited proliferation of human gingival fibroblast and the highest cytotoxicity among tested materials (p<0.05). 3. The Ni-Cr alloy limited the proliferaion of the human gingival fibroblast compared to titanium(p<0.05) but cytotoxicity on the bottom of well was not so considerable, compared to titanium. 4. On the scanning electron micrographs , the ceramic showed good attachment and proliferation of human gingival fibroblast, which was similar to titanium. But gold alloy and Ni-Cr alloy showed the shrinkage of gingival fibroblast both after 24 hours and 3 days. On 5th day, small amount of the human gingival fibroblast proliferation was observed on the Ni-Cr alloy, while the shrinkage of gingival fibroblast was still observed on the gold alloy. Conclusions: These results suggest that the ceramic abutment is as biocompatible as titanium to make proper mucosal seal. The gold alloy has a high cytotoxicity to limit proliferation of gingival fibroblast, which suggest limited use on the anterior tooth where soft tissue healing is recommeded.

• Restorative Dentistry and Endodontics
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• v.24 no.2
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• pp.416-425
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• 1999

The purpose of this study was to evaluate shear bond strength of glass ionomer cements and compomer according to dentin surface treatment method. The materials used in this study were dentin conditioner and cavity conditioner for dentin treatment: Ketacfil, Fuji II LC, and Dyract for restoration. In this study, 90 sound bovine teeth were selected and then the teeth were embeded in improved stone and were grounded with 400 to 600 grit silicon carbide paper to create a flat dentin surfaces. The teeth were divided into nine groups as follows ; Group 1A : Samples bonded to dentin surface with Ketacfil after no treatment Group 1B : Samples bonded to dentin surface with Ketacfil after applicating dentin conditioner Group 1C : Samples bonded to dentin surface with Ketacfil after applicating cavity conditioner Group 2A : Samples bonded to dentin surface with Fuji II LC after no treatment Group 2B : Samples bonded to dentin surface with Fuji II LC after applicating dentin conditioner Group 2C : Samples bonded to dentin surface with Fuji II LC after applicating cavity conditioner Group 3A : Samples bonded to dentin surface with Dyract after no treatment Group 3B : Samples bonded to dentin surface with Dyract after applicating dentin conditioner Group 3C : Samples bonded to dentin surface with Dyract after applicating cavity conditioner Treated dentin surfaces were observed under SEM. After filling of each materials, shear bond strenth was evaluated and then debonded surfaces were observed under SEM. The following results were obtained; 1. The shear bond strengths obtained were decreased as Fuji II LC, Dyract, Ketacfil in that order and there was statistically significant difference(p<0.05). 2. About Group 1. the shear bond strengths were decreased as 1C, 1B and 1A in that order. But there was no significant difference between group 1B and 1C (p<0.05). 3. About Group 2, the shear bond strengths were decreased as group 2B, 2A and 2C in that order. And there was significant difference between group 2B and 2C (p<0.05). 4. About Group 3, the shear bond strengths were decreased as group 3A, 3C and 3B in that order. And there was signicant difference between group 3A and 3B (p<0.05). 5. As a result of observation under SEM, the fracture patterns of Fuji II LC and Dyract were adhesive failures, but those of Ketacfil were cohesive failure of material and mixture of cohesive and adhesive failure.

• Journal of Pharmaceutical Investigation
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• v.20 no.4
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• pp.223-228
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• 1990

The contribution of endogenous transport systems to the blood-brain barrier (BBB) transport of basic and acidic drugs was studied by using a carotid injection technique in rats and an isolated bovine cerebrovascular disease state were compared between the normotensive rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) which have been well established as an animal model with pathogenic similarities to humans. Basic drugs such as eperisone, thiamine and scopolamine inhibited, in a concentration dependent manner the in vivo uptake of $[{^3}H]choline$ through BBB, whereas amino acids and acidic drugs such as salicylic acid and valproic acid did not inhibit the uptake. The uptake of $[^3H]choline$ by B-CAP increased with time and showed a remarkable temperature dependency. The uptake of $[^3H]choline$ by B-CAP showed the very similar inhibitory effects as observed in the in vivo brain uptake, and was competitively inhibited by a basic drug, eperisone. The in vivo BBB uptakes of $[^3H]acetic$ acid and $[^{14}C]salicylic$ acid were dependent on pH of the injectate and the concentration of drugs. Several acidic drugs such such as salicylic acid, benzoic acid and valproic acid inhibited the in vivo uptake of $[^3H]acetic$ acid, whereas amino acid, choline and a basic drug such as eperisone did not inhibit the uptake. The uptake of acetic acid by B-CAP was competitively inhibited by salicylic acid. The permeability surface area product (PS) through BBB for $[^3H]choline$ in SHRSP was significantly lower than that in WKY. The concentration of choline in the brain dialysate in SHRSP was about half of that in WKY, while no significant difference was observed in the plasma concentration of choline between SHRSP and WKY. No significant difference was observed in the transport of monocarboxylic acids, glucose and neutral amino acid through BBB between SHRSP and WKY. From these results, it was concluded that BBB transport system of choline contributes to the transport of basic drugs through BBB, that acidic drugs can be transported via a moncarboxylic acid BBB transport system and that the specific dysfuntion of the BBB choline transport in SHRSP was ascribed to the reduction of the maximum velocity of choline concentration in the brain interstitial fluids.

• Journal of Embryo Transfer
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• v.11 no.2
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• pp.125-135
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• 1996

The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.243-248
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• 2003

Sperrnatogenesis, the process by which the male germ-line stem cells(GSCs; type A spermatogonia) divide and differentiate to produce the mature spermatozoa, occurs in the seminiferous tubules of the testis. The GSCs proliferate actively to produce two types of cells: other GSCs and differentiating spermatogonia. GSCs have unipotentcy, devoted solely to the generation of sperm. The function of GSCs has broad implications for development, disease, and evolution. Spermatogenesis is fundamental for propagation of species and the defects of this system can result in infertility or disease. The ability to identify, isolate, culture, and alter GSCs will allow powerful new approaches in animal transgenesis and human gene therapy relating to infertility. Until recently, research on stem cells in the testis has been limited because of technical difficulties in isolating and identifying these cell populations. Here, we were trying to find out optimal conditions for in vitro culture of GSCs for identifying and isolating GSCs. We collected mouse GSCs from 3-days old mouse by two-step enzyme digestion method. GSCs were plated and grown on mouse embryonic fibroblasts in Dulbecco's modified Eagle's medium (DMEM) containing 15％ fatal bovine serum, 10 mM 2-mercaptoethanol, 1％ non-essential amino acids, 1 ng/$m\ell$ bFGF, 10 $\mu$M forskolin, 1500 U/$m\ell$ human recombinant leukemia inhibitory factor (LIF). Over a period 3∼5 days, GSCs gave rise to large multicellular colonies resembling those of mouse pluripotent stem cells. After 5th passages, cells within the colonies continued to be alkaline phosphatase and Oct-4 positive and tested positive against a panel of two immunological markers(Integrin $\alpha$ 6 and Integrin $\beta$ 1) that have been recognized generally to characterize GSCs. SSEA-1, SSEA-3, and SSEA-4 also showed positive signals. Based on our data, these GSCs-derived cultures meet the criteria for GSCs itself and even other pluripotent stem cells. We reported here the establishment of in vitro cultures from mouse male GSCs.

• Journal of Embryo Transfer
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• v.9 no.1
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• pp.95-102
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• 1994

본 연구는 종모우의 선발방법으로 난포란을 이용하여 실험실내 정자의 수정능력을 직접 검정하여 평가코자 시도되었다. 즉 본 실험은 후대검정중에 있는 한우 후보종모우 15두의 동결융해정자의 수정능력을 평가하기 위하여 정액을 고장액(HIS)에 처리한 후 DM에서 6시간 그리고 소 난포액이 20% 첨가된 DM에서 4시간 전배양하여 수정능을 획득시켜 정자의 활력과 첨체 반응율을 조사하였고 전배양된 정자의 체내(토끼 난관) 또는 체외수정능력을 조사하기 위하여 FCS 15%, 발정암소혈정(CSS) 10%가 첨가된 mKRB에서 체외성숙된 한우난포란과 수정시켜 수정능력을 평가하였으며 인공수정에 의한 개체별 수태율과도 비교 검토한 바 다음과 같은 결론을 얻었다. 1. 한우 난포란의 체외성숙율은 BSA 첨가구 에서 43.8%, FCS 15% 첨가구에서 67.4%, CSS10% 첨가구에서 69.9%이었다. 2. 토끼 난관에서 체외수정율은 BSA 참가구에서 43.8%, FCS 15% 참가구 41.2% 및 CSS 10% 참가구 35.0% 이었다. 3. 후보종무우 15두의 정액을 HIS-DM으로 처리후 6시간 전배양하였을 때 정자의 활력지수는 9-32%였고 첨체반응율은 19-44% 이었으며 20% 난포액을 첨가하여 4시간 전배양 하였을 때 정자의 활력지수는 9-13% 이었고 첨체반응율은 20-43%로 개체간에 차이가 있었다. 4. 체외수정율은 6.6-85.7%였으며, 발정암소혈청(CSS) 10%가 첨가된 mKRB에서 성숙시킨 난포란이 FCS 15% 첨가된 mKRB에서 성숙시킨 난포란보다 다소 높았으나, 정자수정능획득방법간에는 차이가 없었 다. 5. 체외수정율에 있어서 전배양후 정자활력지수와는 부의 상관이 었으며, 첨체반응율과는 낮은 정의 상 관을 나타냈다. 6. 종모우의 수태율은 체외수정율, 정자활력지수 및 첨체반응율과 낮은 정의 상관관계를 나타냈다. 7. 종모우의 개체간 수태율 우열순위에서는 수정율순위와의 사이에 더욱 낮은 부의 상관관계를 보였다. 8. 이상의 연구결과 비록 후대정검중의 제한된 자료로 인하여 종모우 수태율과 체외수정율간에 유의적 인 상관관계는 없었으나, 연결 한우 수정율 평가에 대한 실험실내의 검정가능성을 찾을 수 있었다.