• KSBB Journal
• /
• v.23 no.5
• /
• pp.452-459
• /
• 2008

To obtain maximal efficacy with minimal systemic side-effects, many studies have been carried out to achieve the controlled release of 5-fluorouracil (5-FU). In this study, biodegradable poly(L-lactide) (L-PLA) microparticles containing 5-FU were prepared by a process, called aerosol solvent extraction system (ASES), utilizing supercritical carbon dioxide. The effects of various organic solvents, drug/polymer feeding ratio, polymer molecular weight, and blending with the same polymers with different molecular weights on the formation of 5-FU loaded microparticles were investigated under a predetermined operating condition from our previous study. The drug recovery, entrapment efficiency, and in vitro drug release kinetics were determined by HPLC assays. The drug recovery obtained from the ASES process was found to be very high, whereas the drug entrapment efficiency was considerably low in all the experiments due to the poor affinity between L-PLA and 5-FU. These results indicated that the precipitation rate of L-PLA might be quite different from that of 5-FU so that there was little chance to form 5-FU loaded L-PLA microparticles.

• Clinical and Experimental Reproductive Medicine
• /
• v.35 no.1
• /
• pp.39-48
• /
• 2008

Objective: The aim of this study was to investigate whether multipotent germline stem cells (MGSCs) can be established from neonatal mouse testis. Methods: Various cells containing MGSCs were collected from neonatal testis of ICR mice and allocated to plates for in vitro culture. After 7 days in culture, the cells were passed to a fresh culture plate and continuously cultured. From the third or fourth passage, the presumed MGSCs were cultured and maintained on mitomycin C-inactivated STO feeder cells. The MGSCs were cultured in a condition where mouse embryonic stem cells (ESCs) are cultured. Characteristics of the MGSCs were evaluated by RT-PCR, immunocytochemistry, alkaline phosphatase activity, karyotyping, and transmission electron microscopy. Results: Two MGSCs lines were established from 9 pooled sets of neonatal testicular cells. MGSCs colonies were morphologically undistinguishable from ESCs colonies and both MGSC lines as well as ESCs expressed undifferentiated stem cell markers, such as Thy-1, Oct-4, Nanog, Sox2 and alkaline phosphatase. Fine structure of undifferentiated MGSCs were similar to those of ESCs and 60% of MGSCs (12/20) had normal karyotype at passage 10. They were able to form embryoid bodies (EBs) and MGSC-derived EBs expressed marker genes of three germ layers. Conclusion: We could establish the MGSCs from neonatal mouse testis and they were differentiated to multipotent lineages of three germ layers. Molecular characteristics of MGSCs were similar to those of ESCs. Our results suggest a possibility that multipotent stem cells derived from testis, the MGSCs, could replace the ESCs in biotechnology and regenerative medicine.

• Journal of Periodontal and Implant Science
• /
• v.24 no.3
• /
• pp.661-670
• /
• 1994

It is well known that the application of dressings after periodontal surgery have benefits to provide the comforts to patient and to promote the healing process with action of bleeding control and temporary stabilization for the operated mobile teeth. But until recently the relationship between periodontal dressings and cells which are composed of periodontium has not been clear. The purpose of this study was to evaluate the cytotoxic effect of soluble extracts from the four different kinds of periodontal dressings, two of them were eugenol type (K.H.pack, Wondrpak) and the others were non-eugenol type (Coe-pak, Periocare), on the human gingival fibroblasts in vitro. Human gingival fibroblasts were primarily cultured from gingiva around third molar during the extraction for preventive purposes. Extracts solution were prepared with culture medium by means of imersing the consistent size of periodontal dressing made from plastic mold. Cell were inoculated into the 24 well plate with $3\;{\times}\;10^4\;cells/well$ of medium at $37\;^{\circ}C$, 100% of humidity, 5% of $CO_2$, incubator for 24 hours. After discard of the supernatant of medium, those cells were cultured with original, 1/2, 1/5, 1/10 diluted soluble extract for 24, 48 and 72 hours, and counted the number of cells using the hemocytometer at each designed time and concentration. Also, the cytotoxic effect of soluble extract was measured by Wataha's MTT assay method. In briefly, cells were inoculated and cultured into 96 well culture plate with $2\;{\times}\;10^4\;cells/well$ for 24 hours. Soluble extracts were applied to cultured cells and incubated for 48 hours at same condition. $50\;{\mu}l$ of MTT solution and DMSO were added into each well for the detection of absorbance with ELISA reader. The measured data were calculated by value of colorimetric assay for survival rate. The results were as follows ; In the case of eugenol type of dressing, original, 1/2 and 1/5 diluted extracts of K.H.pack showed very low survival rate. And original extract of Wondrpak showed strong cytotoxic effect and 1/2 diluted extract showed moderate cytotoxic effect. In the case of Non-eugenol type of dressings, only original extract of Coe-pak revealed strong cytotoxic effect and Periocare had little cytotoxic effect. It is concluded that eugenol type of dressings showed more cytotoxic effect than non-eugenol types. This study suggest that use of non-eugenol dressings after periodontal surgery is recommended.

• Journal of Veterinary Clinics
• /
• v.23 no.2
• /
• pp.123-128
• /
• 2006

Production of cloned pigs by somatic cell nuclear transfer (SCNT) has unlimited value for developing critical biotechnology such as xenotransplantation. Various efforts have been made to establish this technology, and several litters of live piglets have been produced after transfer of SCNT embryos. However, the efficiency is very low compared to piglet production by artificial insemination or natural mating. So far, most studies have been limited to in vitro production of SCNT embryos. This study was conducted to standardize a surrogate recipient (gilts) for transfer of SCNT embryos to improve pregnancy rate. Potential surrogate gilts over 7 months of age were checked for their estrous status by observing external signs; vaginal fluid, vulva redness, vulva swelling, and standing response to back pressure. Viscosity of vaginal fluid was evaluated and classified as none (0), medium (1), and strong (2). Vulva redness and swelling was respectively assessed by none or shrink (0), medium (1), strong (2). Back pressure was estimated by an immediate move (0), standing less than 10 sec (1), and standing over 10 see (2). And then ovulation status of each surrogate was classified as pre-ovulation (PO-17 surrogates), just prior to ovulation (JPO-20 surrogates), in ovulation (IO-12 surrogates), just after ovulation (JAO-14 surrogates) and after ovulation (AO-24 surrogates) at the time of surgery for embryo transfer (ET). Real-time ultrasonographic scanners have been used for pregnancy diagnosis by observing amniotic vesicles. The first pregnancy diagnosis was done on Day 30 after ET and then repeated 2-week interval. In the results, SCNT embryos transferred into JPO surrogates gave better pregnancy rates (45%) than others (4% to 11%) on Day 30 after ET. These result indicates that surrogate gilts in a status just prior to ovulation can offer optimal condition to establish pregnancy by transfer of SCNT pig embryos.

• The Journal of Korean Academy of Prosthodontics
• /
• v.45 no.3
• /
• pp.303-309
• /
• 2007

Statement of problem. The effect of gold electroforming on gold alloy was not studied. Purpose. This in vitro study investigate the effect of gold electroforming on gold-silver-palladium alloy. Material and methods. Three pieces of gold strips had undergone the electroforming procedures on one side and then half of the side again electroformed. The set mode for this study was program 1 ($200{\mu}m$). And the processing time was 15min (1/20 time to form $200{\mu}m$ coping). The confocal laser scanning microscope (PASCAL 5, Carl Zeiss, Bernried, Germany) was used to measure the thickness of the pure gold layer electroformed on the gold strips. Half of the gold strip was coated two times with electroformed gold, and the other half one time. The data from the cone focal laser system was processed to get the vertical profile of the strips and the difference of the vertical height between the double coated and single coated layer was regarded as the thickness of the gold coating. The layer thickness value to built 3D image of the cone-focal laser was set $0.5{\mu}m$. Next to the measurement of the thickness of the coating, the Vicker's hardness test was done. It was performed on the double coated surface, single coated surface and non-coated surface (back side) three times each. Results. The mean thickness value gained from gold electroforming technique was measured to be $22{\mu}m$ for sample 1, $23{\mu}m$ for sample 2, $21{\mu}m$ for sample 3. In the same condition of time, power and the amount of electrolyte, the data showed no difference between samples. According to the results of variance analysis, the differences among the variations in number of coating were statistically insignificant (p>0.05), meaning that the two times of gold electroforming coating did not change the hardness of gold-silver-palladium alloy. Conclusion. The test of thickness of gold coating proved the coherency of the gold electroforming procedure, in other words, when the power, the exposed surface area, processing time and the amount of electrolytes were set same, the same thickness of gold would be coated on. The hardness test showed that the electroformed gold coating did not change the hardness of the gold-silver-palladium alloy when it is coated not more than $45{\mu}m$.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.25 no.4
• /
• pp.403-415
• /
• 2009

The purpose of the study is to establish the effect of different abutment shapes and types of cements on the retention of the full veneer casted gold crowns. Metal dies that has the similar shape with the implant abutment were manufactured using a short(5mm) and long(10mm) dies with different convergence angles. Metal dies and gold crowns, which were made from the metal dies, were cemented with Temp-bond, Temp-bond mixed with petroleum jelly, ZPC and Premier implant cement. After that, these were tested for tensile force at the point of separation. The effect of convergence angle changes of different cement types on the retention was studied as well as the effects of the cement type changes with different convergence angles on the retention. In addition, study about the marginal gap of Premier implant cement used for this experiment was conducted. The results are as followed under the in-vitro experimental limits; 1. The retention of the Temp-bond mixed with petroleum jelly decreased as the convergence angle increased, and the retention was weakest among the cements. 2. The retention of ZPC decreased as convergence angle increased. When convergence angle was 5 degrees, ZPC showed stronger retention than Premier implant cement. 3. Premier implant cement had the weakest retention when the convergence angle was 5 degrees but when the convergence angle was 10 degrees, it had the strongest retention. As the angle increased more than 10 degrees, the retention decreased. 4. Premier implant cement showed bigger marginal gap when the convergence angle was 5 degrees than 10 degrees under the experimental condition.

• Research in Plant Disease
• /
• v.17 no.3
• /
• pp.381-385
• /
• 2011

Effects of temperatures, relative humidity, pH, and dry periods on conidial germination of Alternaria dauci were evaluated under in vitro conditions. Conidial germination was accelerated at over 95% relative humidity in $15^{\circ}C$ to $25^{\circ}C$ condition. Conidial germination was rapidly reduced at 5 regardless of relative humidity conditions. More than 50% of the conidial germination were initiated within 2 h at $25^{\circ}C$ through pH 5 to 7. The highest conidial germination of A. dauci was on 0.2% of carrot leaf extract. Conidia could survive longer than 12 h, even though its germination decreased. After a 12 h dry period, around 10% of conidia revived and germinated when conidia were hydrated again. These results could be used as the useful information on conidial germination of A. dauci and ecology of Alternaria leaf blight.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.17
• /
• pp.7749-7754
• /
• 2015

Background: Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Materials and Methods: $Lipofectamine^{TM}2000$ as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA, miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. Results: The seeding density of Hela cell line and Siha are $1.5{\times}10^4$ (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (P<0.01). MTT assay showed that according to the above conditions which has the lowest cytotoxicity. Conclusions: The method of Liposome to transfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.

• Journal of Food Hygiene and Safety
• /
• v.30 no.3
• /
• pp.295-301
• /
• 2015

The functions of probiotics, particularly Lactic acid bacteria, have been studied in a range of human diseases, including cancer, infectious diseases, gastrointestinal disorders, and allergies. Among the many benefits associated with the consumption of probiotics, modulation of immune activity has received the most attention. This study aimed at investigating the improved immune stimulatory and stability of L. plantarum when cultivated on modified basal media supplemented with the Undaria pinnatifida co-cultured with L. plantarum. An in vitro test showed that U. pinnatifida media cultured L. plantarum is strong enough to survive in the gastric juice (gastric and bile acid). Mouse macrophage-derived cell lines RAW 264.7 was used to measured immune stimulating activity of L. plantarum. When U. pinnatifida media cultured by L. plantarum was NO and $TNF-{\alpha}$ production is significantly increased compared to basal media cultured L. plantarum. These results show that U. pinnatifida could be applied for a component for cultivation of L. plantarum. This optimized U. pinnatifida medium can be used the improving of stability and immune function on production of probiotics.

• Journal of Korean Society of Forest Science
• /
• v.73 no.1
• /
• pp.33-42
• /
• 1986

This study was carried out to investigate the optimum conditions for isolation and culture of mesophyll protoplasts from Populus alba ${\times}$ P. glandulosa. The results obtained from the experiments are as follows; 1) The suitable concentration of BAP for shoot multiplication was 0.4 mg/l. 2) High yield and viability of isolated protoplasts were obtained by our high enzyme-short time incubation method. 3) Optimum enzyme concentrations for mesophyll protoplast isolation were Cellulase 2%, Macerozyme 0.8%, Hemicellulase 1.2%, Driselase 2%, and Pectolyase Y-23 0.05%. 4) 0.6M mannitol in enzyme solution was the most effective for protoplast isolation and viability. 5) The most adequate pH level of enzyme solution was pH 5.6. 6) The effect of DTT and MES buffer was significant. 7) For protoplast purification, 0.6M sucrose was the most proper concentration. 8) The adding effect of Dextran T40 in floating solution was important. 9) The mesophyll protoplasts isolated through our high enzyme-short time incubation method revealed successful response to culture condition over 3 weeks of culture.