• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.4
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.

• Tuberculosis and Respiratory Diseases
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• v.53 no.4
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• pp.389-400
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• 2002

Background : The rapid diagnostic tests for tuberculosis are needed to facilitate early treatment of tuberculosis and prevention of Mycobacterium tuberculosis transmission. The Xeniss Rapid TB kit is a rapid, card-based immunochromatographic test for the detection of antibodies directed against M. tuberculosis antigens including antigen 5(38-kDa antigen). The objective of this study was to evaluate the performance of the Xeniss Rapid TB kit for the diagnosis of active tuberculosis with serums from patients, asymptomatic healthy and close contact controls. Methods : 188 patients with active tuberculosis were tested; 177 with pulmonary tuberculosis(18 with combined pleurisy), and 11 with extrapulmonary tuberculosis. The control groups were composed of 82 close contacts and 57 healthy adults. Study subject were drawn from one national tuberculosis hospital for patients and close contacts, and another private hospital for healthy adults in Masan city, Korea. The Xeniss Rapid TB kit(Xeniss Life Science Co., Ltd., Seoul, Korea) was evaluated by using serum samples according to the instructions of the manufacturer by an investigator masked to the clinical and microbiological status of the study subjects. Results : The diagnostic sensitivity of the Xeniss Rapid TB kit was 73.9% in patients and specificities were 73.2% and 93.0% in close contact and healthy adults respectively. The positive predictive value in patients was 84.2% and the negative predictive value in controls was 85.8%. Conclusion : This study shows that the Xeniss Rapid TB test is a simple and fast method to diagnose active TB. The results of the sensitivity and specificites suggest that serodiagnosis using this point of care testing(POCT) device would be valuable and advantageous for screening tuberculosis in the clinical field.

• Korean Journal of Poultry Science
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• v.42 no.2
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• pp.147-156
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• 2015

This study was conducted to investigate the effects of dietary resveratrol on growth performance, blood biochemical parameters, immunoglobulin, and blood antioxidant activity in broiler chicks. Three hundred twenty one-day old broiler chicks were divided 8 treatments (C(-), basal diet; C(+), basal diet with antibiotics; DL-${\alpha}$-tocopherol 20 IU; DL-${\alpha}$-tocopherol 200 IU; resveratrol 20 ppm; resveratrol 200 ppm; methylated resveratrol 20 ppm; methylated resveratrol 200 ppm) with 4 replicates and 10 birds per replicate. Birds were reared for 35 days, and, at the age of 35 days, eight birds of average weight from each replicate were selected for blood samples collection. There were no significant differences on feed intake and feed conversion ratio. But final body weight and weight gain in antibiotics, resveratrol and methylated resveratrol treatments were significantly higher than no-antibiotics and ${\alpha}$-tocopherol treatments (P<0.05). There were no significant differences on carcass rate and relative organ weights among treatments, however, weights of liver and bursa of februcius in antibiotics, resveratrol and methylated resveratrol treatment were lower than other treatments. Weight of pancreas was high in resveratrol and methylated treatment. On the cecal microflora (total microbes, Coliform bacteria, Salmonella spp., and lactic acid bacteria), these in resveratrol and methylated resveratrol treatments didn't show the differences compared with those in no-antibiotics, antibiotics, and ${\alpha}$-tocopherol treatments. In the serum, there were no significant differences on creatinine, blood urea nitrogen (BUN), total protein, albumin, globulin, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) among treatments, though globulin contents of reseveratrol 200 ppm and methylated resveratrol 20 ppm treatments decreased compared to those of other treatments. Immunoglobulin (IgA, IgG and IgM) were significantly decreased in antibiotics and resveratrol treatments compared to that of no-antibiotics and ${\alpha}$-tocopherol treatments (P<0.05). Superoxide dismutase (SOD) like activity tended to increase in resveratrol groups (P<0.05), however, there was no significant difference on malondiakdehyde (MDA) content among treatments. In conclusion, these results showed that resveratrol derived from mulberry can be used as alternative of antibiotics through improvement of broiler's performance and maintain of health.

• Pediatric Infection and Vaccine
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• v.5 no.2
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• pp.239-244
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• 1998

Purpose : The incidence of hepatitis A virus infection in Korea has decreased recently. However, the prevalence in children and young adults is on an increasing trend in Inchon-Kyunggi prefecture. Economic development that leads to better living conditions and improves hygiene facilities has reduced the rate of HAV infection. This study was designed to evaluate the recent seroprevalence rate of hepatitis A in children and adolescents. Methods : Study population consisted of 612 children and adolescents aged 1 to 17 years in Inchon-Kyunggi prefecture. Serum samples were collected between 1996 and 1997 and tested for anti HAV by Microparticle Enzyme Immunoassay(MEIA). Results : According to age, the seropositive rates were 10.5%(male 13%, female 6.7%) in 1 year-old group, 0% in 3, 5, 11 and 13 year-old group, 6%(male 8%, female 4%) in 15 year-old group, and 11%(male 14%, female 8%) in 17 year-old group. Conclusion : This study showed that seropositive rate of HAV infection increased with age. While it revealed extremely low incidence of HAV infection in children less than 13 years old, they are also likely 10 be exposed 10 the infection. Our findings suggest that the effective administration of vaccination is needed in selected and high risk groups.

• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.3
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• pp.441-446
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• 2001

Flumazenil is a competitive antagonist of benzodiazepines. It is usually administered intravenously. However, if the intravenous route is not available then other routes of drug administration should be considered. This study was designed to evaluate the reversal effects of flumazenil after nasal administration. Twenty-five young, healthy adult volunteers participated in this clinical trial. The dosage of 0.08mg/kg midazolam was administered intravenously to induce deep sedation. Ten minutes after midazolam administration, 0.5mg of flumazenil was dropped nasally, over a period of one minute. Blood samples were taken to measure the concentration of midazolam and flumazenil at 0, 5, 10, and 20min after nasal administration of flumazenil, using High Performance Liquid Chromatography. The degree of sedation was evaluated with sedation score and bispectral index (BIS), Statistical analysis was performed by multivariate ANOVA and correlation analysis (P<0.05). Peak serum flumazenil concentration was reached in 10min. Sedation score decreased after midazolam administration and showed a significant increase after flumazenil administration. However, BIS decreased during the first 10min after midazolam administration and then no significant changes after flumazenil administration. There were two instances representing rapid and complete reversal of midazolam after intranasal administration of flumazenil. In conclusion, intranasal flumazenil administration may be effective in some patients when intravenous route is not available in condition of benzodiazepine overdose.

• Journal of the Korean Society of Food Science and Nutrition
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• v.5 no.1
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• pp.87-92
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• 1976

Comparing the amount of Hb, the nutriture of the deaf-mutes are better than that of the amentias and the nursery school children. The former, however, shows less condition than that of the boys and girls in the junior physical education school (Hb : 12.7) and the normal family children (Hb : 10.8). In the group of six to eleven years old that is a class a elementary school, the physical condition such as stature, chest circumference, and weight of the deaf -mute is respectively almost the same degree, which is above standard in our country. On the contrary the degree of the amentias and the nursery school children has an inferiority approaching to the standard. In the group of twelve to fifteen years old that is a class of junior high school, the nursery school children have the lowest physical condition and the boys and girls of the physical training school the highest. The order of the growth of physical condition is as follows : Nursery school children

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

• Korean Journal of Animal Reproduction
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• v.9 no.1
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• pp.9-30
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• 1985

The study was carried out to find out the changes of the sex hormone levels in the milk of Holstein cows during the reproductive stages such as the estrous cycle, pregnancy and periparturient period. The FSH, LH, estradiol-17$\beta$ and progesterone from the milk samples were assayed by radioimmunoassay methods. The results of this study were summarized as follows: 1. The levels of progesterone and estradiol-17$\beta$ were similar among inter-quarters, but they were higher in after milking than before milking times, with no statistical significance. 2. The milk progesterone levels during the estrous cycles reached a peak mean level of 3.55$\pm$0.26ng/$m\ell$ at 15 days after estrus and they did not show any differences among the length of estrous cycles. The estradiol-17$\beta$ levels during the estrous cycles showed a peak level of 36.40$\pm$2.38pg/$m\ell$ at estrus, and decreased(17.20$\pm$0.46 pg/$m\ell$ to 18.65$\pm$1.26pg/$m\ell$) at luteal phase. 3. The FSH levels during the estrous cycles ranged from 2.25$\pm$0.23mIU/$m\ell$ to 4.35$\pm$0.24mIU/$m\ell$ showing significant changes. The LH levels during the estrous cycles gradually increased and remained a peak level of 10.90$\pm$0.36mIU/$m\ell$ from 20 to 25 days after estrus. 4. The progesterone levels during the pregnancy were decreased from 30 to 60 days after artificial insemination, and therafter continuously increased until 240 days. The estradiol-17$\beta$ levels during the pregnancy were 24.56$\pm$1.19pg/$m\ell$ at day 30 after artificial inseminaton, and increased rapidly until 180 days. The levles were agagin decreased by 26.17$\pm$3.03pg/$m\ell$ until 210 days and markedly increased by 68.00$\pm$8.70pg/$m\ell$ until 240 days. 5. The prolactin levels during the pregnancy were 31.27$\pm$2.31ng/$m\ell$ and 42.60$\pm$2.37ng/$m\ell$ at day 150 and 240 after artificial insemination respectively. The LH levels during the pregnancy reached a peak of 27.47$\pm$7.90mIU/$m\ell$ at day 30 after artificial insemination, and thereafter gradually decreased. 6. The progesterone levels during the periparturient period reached a peak of 4.61$\pm$0.34ng/$m\ell$ at day 3 prepartum, and thereafter gradually decreased, and showed 2.05$\pm$0.60ng/$m\ell$ at day 7 postpartum. The estradiol-17$\beta$ levels during the periparturient period showed high level from 207.23$\pm$6.04pg/$m\ell$ at day 1 prepartum to 239.90$\pm$13.90pg/$m\ell$ at day 2 prepartum, and thereafter began to decline and reached 51.87$\pm$1.72pg/$m\ell$ at by 7 postpartum. 7. The prolactin levels during the periparturient period showed relatively higher level at the time of parturition. The LH levels during the periparturient period rnage from 6.32$\pm$0.32mIU/$m\ell$ to 13.90$\pm$1.37mIU/$m\ell$ showing significant changes. 8. The progesterone levels(4.6$\pm$0.8ng/$m\ell$) of the pregnant cows were significantly higher than those (1.84$\pm$1.4ng/$m\ell$) of nonpregnant cows. The cows of artificial insemination from 61 to 90 days after parturition showed higher progesterone levels. 9. During 20 to 25 days after artificial insemination, the accuracy of pregnancy diagnosis from milk progesterone levels were 94.4% for nonpregnant cows(<2.3ng/$m\ell$), and 75.0% for pregnant cows( 3.2ng/$m\ell$). The average overall accuracy of pregnancy prediction for nonpregnant and pregnant cows 83.3% 10. The results obtained this study suggest that the understanding of the endocrinological mechanisms by means of milk hormone analysis during the estrous cycle, pregnancy and parturition would give the basic information needed for increasing efficiency of reproduction. This study would not only provide an accurate method of the early pregnancy diagnosis by milk progesterone levels but also contribute to the research of providing the method of detecting of FSH levels in milk, which was difficult in blood serum.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.469-490
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• 1996

The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.