• 대한한방내과학회지
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• 제31권2호
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• pp.189-200
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• 2010

Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

• 약학회지
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• 제22권1호
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• pp.33-41
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• 1978

In order to study the growth and survival of enteric pathogens causing water-borne infections in sewage, the filter-sterilized and autoclaved sewages of Dae Gu City were inoculated with Salmonella typhimuriuim, Shigella flexneri 2a, Sh. sonnei I, Vibrio eltor and V. parahaemolyticus, as test series and Escherichia coli as control. After varying periods of incubation up to 15 days at $4^{\circ}$, $15^{\circ}$, $25^{\circ}$ and $37^{\circ}C$, viable cells in the inoculated sewages were counted by colony count technique. Distilled water and 0.9% saline were subjected to inoculation of the organisms was observed in the filter-sterilized and autoclaved sewages at $4^{\circ}$ and the sewages became sterile within a few days. At $15^{\circ}$, no growth and rapid inactivation of the organisms in the filter-sterilized sewage and slight or no growth in the autoclaved sewage was noted. Some viable cells were found in the autoclaved sewage after 15 days. A considerable growth was observed in the filter-sterilized and autoclayed sewages, at $25^{\circ}$ and $37^{\circ}$, and large numbers of viable cells were found even after 15days of incubation. In general, the autoclaved sewage supproted the growth more noticeably than the filter-sterilized, except for V.parahaemolyticus which grew well in filter-sterilized sewage. No marked difference was noted between incubations at $25^{\circ}$ and $37^{\circ}$, but V. parahaemolyticus showed a slightly more active growth at $25^{\circ}$ than at $37^{\circ}$. Distilled water inactivated the organisms within a few days, but saline supported the growth at $25^{\circ}$ and $37^{\circ}$. Marked differences were noted in the survival test of sewages pathogens of different origins.

• 대한수의학회지
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• 제33권3호
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• pp.429-442
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• 1993

Serpulina(S) hyodysenteriae B 204 strain이 생산한 세포독소에 의한 장점막의 변화를 알아보기 위하여 일반사육조건에서 사육된 8주령, 수컷 잡종돼지 2마리의 결장을 외과적으로 결찰한 계제(loop)를 이용하여 실험하였다. Serpulina균을 trypticase soy broth(TSB)에 배양하여 결찰한 결장계제 4곳에 각각 멸균한 Serpulina균 TSB, 여과한 Serpulina균 TSB, 세척한 Serpulina균, 무처치 Serpulina균 TSB를 접종하였다. 점막조직을 접종(p.i.) 24, 48시간 후에 채취한 후 처리하여 주사전자현미경과 투과전자현미경으로 관찰하였다. 여과한 Serpulina균 TSB를 접종한 경우는 변화가 관찰되지 않았으나 세척한 Serpulina균을 접종한 경우는 무처치 Serpulina균 TSB를 접종한 부위에서 나타난 것과 유사한 초기변화가 관찰되었다. 본 실험의 결과는 Serpulina균의 세포독소가 실험적 감염시 초기병변형성에 별로 기여하지 않음을 시사하였다. Serpulina균의 독소가 병변을 야기하는데 관여하는 기전은 밝혀지지 않았다.

• 한국동물위생학회지
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• 제34권2호
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• pp.159-166
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• 2011

This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

• Journal of Periodontal and Implant Science
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• 제25권3호
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• pp.539-556
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• 1995

Periodontal therapeutic modalities should be re-establishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, periodontal ligament cells must selective migrate to the deneded root surface, attached and proliferated it. Local pH concentration is one of the most factors that periodontal regeneration. The aims of this study were to examine on biological effects of pH to the human periodontal ligament cells in vitro, especially on the cell morphology, attachment, activity, vitality and viability. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Immediately after extraction, any soft tissue adhering to the cervical parts of the roots was carefully removed with a sterile curette. To produce different pH levels in the media, Eagle's MEM was adjusted from pH 6.6 to 8.2 in 0.2 intervals with 1 M NaOH and 1 N HCl. After cultivation, Then, Periodontal ligament cells were cultured at pH ranging from 6.6-8.2. attachment assay was done at 1, 2 day incubation and activity assay was done at 1, 2, 3 day incubation. The experiments were evaluated by scaning electron microscopic techniques (HITACHIX-650 Scaning Electron Microanalyzer, Tokyo, Japan), MTT assay, and the cultured periodontal ligament cells were fixed in neutral formalin for 24 hours and immunohistochemically processed by PCNA for proliferating ability. The surviving cells in the medium showed slightly increased volume and widening intercellular distances at low concentration of pH than control group (pH 7.4), and apparently shrinkage at high concentration of pH than control group (pH 7.4). The results of the statistical analysis from the experiment on attachment, vitality and viability were as follows. Attachment of periodontal ligament cells at 1st and 2nd day, similar attachment rate of low concentration pH compared with control value (pH 7.4). But above pH 8.0, attachment rate were statistically significant decrease from control value(P<0.05). Periodontal ligament cell's activities were maximum at pH 7.6 by MTT assay. Similar with control value at low concentration of pH. But, the activities were statistically significant decrease at high concentraration of pH(P<0.05). Cellular proliferating rate (PCNA index) were statistically significant decrease from control value at low and high concentration of pH(p<0.05). This results suggested that hjgh concentration pH, in other words, alkali pH was cytotoxic effects on human periodontal ligament cells in vitro.

• 한국동물위생학회지
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• 제42권4호
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• pp.257-264
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• 2019

Mastoparan V1 was used as adjuvant of formalin-inactivated Salmonella Gallinarum whole cells vaccine against fowl typhoid in a chicken model. The 75 brown nick chickens were equally divided into 5 groups, and all chickens of each group were immunized at 6 weeks of age (0 WPPI; weeks prime post immunization), and at 9 weeks of age (3 WPPI) (except group B). Group A chickens were intramuscularly (IM) inoculated with 500 uL of sterile phosphate-buffered saline (PBS), and group B chickens were subcutaneously immunized with 0.2 ml containing 5×10⁷ viable vaccine strain/bird. The chickens in groups C~E were IM inoculated with approximately 3×10⁹ cells/0.5 mL of formalin-inactivated the S. Gallinarum whole cells, approximately 3×10⁹ cells/0.5 mL of formalin-inactivated the S. Gallinarum whole cells with mastoparan V1 as adjuvant, and 0.5 mL of PBS, respectively. S. Gallinarum outer membrane proteins-specific serum IgG titers were considerably higher in groups B~D than in groups A and E. However, the levels of IFN-γ in groups B and D only than in groups A and E were significantly higher. Following oral challenge with virulent wild-type S. Gallinarum, no chicken in groups A (no challenge group) and B was dead, and only 30% of chickens in group D was dead. However, 70% of chickens in group C and all chickens in group E were dead after oral challenge. The results of this study demonstrated that IM immunization with approximately 3×10⁹ of the formalin-inactivated S. Gallinarum whole cells containing mastoparan V1 induced robust antibody and cell-mediated immune responses in chickens. The whole cells also conferred protection against infection with wild-type S. Gallinarum.

• 대한수의학회지
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• 제30권4호
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• pp.447-457
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• 1990

In order to observe the distribution of mast cell on the stages of the mammary carcinogenesis, the numerical changes of mast cells in the mammary tumor development in rats treated with DMBA and compound 48/80 have been investigated by the light microscope. The results observed were summarized as follows: The appearance of tumor were not observed during the whole experimental period in the rats of the control group received injection of sterile saline, but tumors appeared in 100% of the animals, the tumor induction time that represented the number of days elapsing between the 3rd DMBA administration until a first tumor became $10{\times}10mm$ in diameter was $42.5{\pm}4.7$ days and the mean number of tumor masses per rat was $3.4{\pm}1.2$ in the DMBA treated group. And the majority of the DMBA-induced mammary neoplasms were appeared cervical mammary gland and thoracic mammary gland. The histological findings of mammary carcinoma were recognized adenocarcinoma in the DMBA treated group. Mast cells were distributed within the adipose tissues and the interglandular connective tissue in the control, but found to be randomly dispersed within the tumor cell masses, in the connective tissues adjacent to the periphery of the tumor, the adipose tissues and the subcutaneous tissues contiguous to the region of tumor development in the DMBA treated group. Numerical alterations of mast cells were observed in the mammary tumors that separated into three major classes of tumors: hyperplasia, atypical hyperplasia and carcinoma. The number of mast cells were distributed in the connective tissues adjacent to the mammary gland was $45.3{\pm}3.4$ cells in the control group, but was $50.2{\pm}4.9$ cells, $126.7{\pm}10.5$ cells and $340.3{\pm}19.2$ cells according to each stages of mammary tumorigenesis in the DMBA treated group.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제17권4호
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• pp.331-336
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• 1995

To use cultured mucosa as a graft of full thickness, our laboratory has been involved in the development of techniques to grow epidermis together with connective tissue. Human oral mucosa was obtained at dental surgery. Under sterile conditions the tissues were cut into explants of 0.1 $cm^2$ which were placed in the center of 24 well tissue culture dishes and incubated in a growth medium. The growth medium used for epithelial was MEM(Minimum Essential Medium) supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide, glutamine (0.292 g/l), epidermal growth factor (40 ug/ml), cholera toxin (30 ng/ml), hydrocortisone (2 ug/ml), insulin (40 ug/ml) and transferin (5 ug/ml). The medium for stratification of epithelial cells was MEM supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide and glutamine (0.292 g/l). The medium used for fibroblasts was MEM supplemented with 10% fetal calf serum. With the three types of media used alternatively, a mucosa composed of epidermis and connective tissue was obtained after 3 weeks of culture.

• 한국환경과학회지
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• 제18권1호
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• pp.9-14
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• 2009

Pfiesteriaand Pfiesteria-like organisms were reported to be linked to major fish kills(involving well over a billion fish) in North Carolina and Maryland estuaries on the U.S. east coast during the 1990s. Occurrences of these species have been recently reported from Korean waters including Chinhae Bay and the coast of Yeosu. In this study, the life cycle of Cryptoperidiniopsis brodyi and Pfiesteria piscicida were examined using DAPI staining. Their excystment and growth were stimulated directly by the addition of prey cells such as Rhodiminas salina. Amoeboid stages in C. brodyi and P. piscicida were never observed in culture, even after addition of filter-sterile fish mucus and tissue. The dominant life cycle stages consisted of motile flagellated zoospores and cysts. A typical dinoflagellate life cycle was demonstrated by direct observation and DAPI staining.

• Animal cells and systems
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• 제4권2호
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• pp.109-115
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• 2000

To clarify taxonomic status of the two sibling species, Moroco oxycephalus and M. lagowskii reproductive isolation mechanisms were investigated at sympatric area located in Kansung-up, Kosung-gun, Kangwon-do, Korea. Genetic analysis was performed to reveal mating system and intensity of Hybridization between the two species. The frequencies of hybrids were increased since 1989, and then the observed hybrid frequencies ($H_O$) did not significantly differ from the expected hybrid ($H_E$) in 1998 and 1999. However, based on histological analysis of two parents and their hybrid s gonads, the hybridizations between M. oxycephalus and M. lagowskii produced mostly fertile females but sterile males in accordance with Haldane s rule. Although it was suspected that pre- and postmating isolation mechanisms were affected between the two species, M. oxycephalus and M. lagowskii seemed to be strongly isolated with microhabitat at sympatry until 1997. Since 1998, hybrid frequencies were increased by habitat disturbance. However, their hybrid frequencies would be reduced by postmating isolation mechanisms. Therefore, the two species are considered to be distinct species recently diverged.