• Title/Summary/Keyword: in-vitro

Search Result 16,409, Processing Time 0.06 seconds

Alteration of Biosynthesis and Secretion of Adrenal Catecholamines in Cycling Rat (발정주기 중 흰쥐 부신에서의 카테콜아민 합성과 분비 변화)

  • 이성호
    • Development and Reproduction
    • /
    • v.6 no.2
    • /
    • pp.105-110
    • /
    • 2002
  • Numerous hormones are involved in the regulation of reproduction. Among them, estrogen and progesterone are the most important ovarian steroid hormones regulating female fertility. On the other hand, diverse stressors impede female receptivity and fertility. Since norepinephrine(NE) and epinephrine(E) are released from the adrenal during stress, it might play a role in stress-induced disruptions of fEmale reproductive parameters. The present study was performed to analyze the changes in adrenal catecholaminergic activities in cycling rats. The tissue content and secretion level of catecholamines were determined by high performance liquid chromatography coupled with electrochemical detector(HPLC-ECD). Adrenomedullary content of norepinephrine(NE) was increased on proestrus stage (59.47 $\pm$ 6.86 ug/gland), peaked on diestrus I stage(65.22 $\pm$ 5.99 ug/gland), and was nadir on diestrus II stage(41.63 $\pm$ 1.33 ug/gland). The highest E content was observed on proestrus stage(361.86 $\pm$ 15.58 ug/gland) while the lowest level was on diestrus II stage(285.58 $\pm$ 12.25 ug/gland). In addition to these observations, a significant reduction of the NE : E ratio was observed (1 : 4.81 on diestrus I vs 1 : 6.13~7.02 on other stages). In vitro secretion of adrenal NE and E was increased on proestrus stage, peaked on estrus stage, and decreased on diestrus II stage. Interestingly, the NE : E ratio in conditioned media was significantly increased on estrus stage (1 : 3.32 vs 1 : 2.34~2.65 on other stages. The biosynthesis of NE and E is mediated by tyrosine hydroxylase(TH) and phenylethanolamine-N-methyltransferase(PNMT) which acts conversion of tyrosine into DOPA and NE into E, respectively. These finding demonstrated that sex steroids, during setrous cycle, seem to be able to modify the adrenal catecholamines biosynthesis and secretion with stage-specific manner by modulation of the enzyme activities.

  • PDF

Effect on the Inoculation of Bacillus on the Growth of Chinese Cabbage and Sesame and on Microbial Flora in Soils (Bacillus subtilis 접종이 배추 및 참깨의 생장(生長)과 토양(土壤) 미생물상(微生物相)에 미치는 영향(影響))

  • Kim, Kwang-Sik;Lee, Jae-Pyeong;Kim, Yong-Woong;Rhee, Young-Hwan;Kim, Yeong-Yil
    • Korean Journal of Soil Science and Fertilizer
    • /
    • v.26 no.4
    • /
    • pp.271-277
    • /
    • 1993
  • An antagonistic bacteria was isolated from rhiaosphere of pepper and corn and identified as Bacillus (B.) subtilis. These B. subtilis B-5 was transformed and marked with the plasmid pCPP4 which possess neomycine resistan. gene. The marked stranins showed growth inhibition to Rhizoctonia (R.) solani, Fusarium (F.) solani, and F. oxysporum in vitro, and were used in studying growth promoting effects on sesame and cabbage. All the identified strains utilize glucose, sucrose, fructose, lactose, mannitol and sorbitol as carbon source, but not rhamnose, and the marked strains also showed characteristics similar to wild-type strains. Germination rate of chinese cabbage and sesame seeds was increased by about 10% or more in the plot to which these strains were inoculated and the effect was higher in soil than in petri dish. The early growth promoting effects of these strains appeared higher, as compared with control plot, in the plots to which B. subtilis B-5 and pathogenic fungi was inoculated together. When the marked strains, B. subtilis B-5NEOr, were inoculated in the rhizosphere of chinese cabbage and sesame with $1.1{\times}10^8CFU/g$ dry soil, the number of inoculated strain was decreased slowly to the level of $10^5{\sim}10^6CFU/g$ dry soil after 4 weeks and the number of Pseudomonas spp. maintanied the level of $10^5CFU/g$ dry soil throught total period, but the number of fungi was decreased rapidly from the early level of $10^8CFU/g$ dry soil to $10^3CFU/g$ dry soil after 4 weeks.

  • PDF

Differential Effects of Green Tea Polyphenol in the ${\gamma}-irradiation$ Induced Human Leukemic and Lymphoblastic Cell Damage (녹차 폴리페놀이 감마선조사에 의한 백혈병과 림프구모세포의 손상에 미치는 영향의 차이)

  • Jeong, Hwan-Jeong;Kim, Eun-Mi;Min, Jung-Jun;Bom, Hee-Seung;Kim, Young-Ho;Jeong, Young-Do;Kim, Chang-Guhn
    • The Korean Journal of Nuclear Medicine
    • /
    • v.37 no.5
    • /
    • pp.308-316
    • /
    • 2003
  • Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

Anti-Allergic Activities of Fermented Eriobotrya japonica and Saurus chinensis Extracts in 2,4-Dinitrochlorobezene-Induced BALB/c Mice (2,4-Dinitrochlorobenzene으로 유도된 아토피 피부염 동물모델에서 비파엽 및 삼백초 추출발효물의 항아토피 활성)

  • Choi, Myung-Jin;Jung, Hee-Kyoung;Jeong, Yoo-Seok;Park, Seung-Chun;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.39 no.11
    • /
    • pp.1611-1618
    • /
    • 2010
  • This study was conducted to evaluate the effects of Eriobotrya japonica and Saurus chinensis extracts and their fermented extracts on immune parameters in BALB/c mice treated with 1% 2,4-dinitrochlorobenzene (DNCB). The groups were Eriobotrya japonica extract (BI), Saurus chinensis extract (SA), mixture with E. japonica extract and S. chinensis extract (FB) and fermented mixture with E. japonica extract and S. chinensis extract (FA) and distilled water treated control. The level of serum immunoglobulin (Ig) E was decreased in FA compared to control group, but significant difference was not observed (p<0.05). The histamine and contents in FA and control group were $1.47{\pm}0.20$ ng/mL and $1.90{\pm}0.04$ ng/mL, respectively (p<0.05). Ceramide contents were significantly increased in FA compared to BI, SA, FB and control group (p<0.05). These results suggest that FA supplementation in the DNCB treated BALB/c mice affect anti-allergic activities positively, and may be used as functional material for suppression of atopy dermatitis in food industry.

Biocompatibility and Histopathologic Change of the Acellular Xenogenic Pulmonary Valved Conduit Grafted in the Right Ventricular Outflow Tract (우심실 유출로에 이식한 무세포화 이종 폐동맥 판막도관의 생체 적합성 및 조직병리학적 변화양상에 대한 연구)

  • 허재학;김용진;박현정;김원곤
    • Journal of Chest Surgery
    • /
    • v.37 no.6
    • /
    • pp.482-491
    • /
    • 2004
  • Background: The xenogenic or allogenic valves after in Vitro repopulation with autologous cells or in vivo repo-pulation after acellularization treatment to remove the antigenicity could used as an alternative to synthetic polymer scaffold. In the present study, we evaluated the process of repopulation by recipient cell to the acellu-larized xenograft treated with NaCl-SDS solution and grafted in the right ventricular outflow tract. Material and Method: Porcine pulmonary valved conduit were treated with. NaCl-SDS solution to make the grafts acellularized and implanted in the right ventricular outflow tract of the goats under cardiopulmonary bypass. After evaluating the functions of pulmonary valves by echocardiography, goats were sacrificed at 1 week, 1 month, 3 months, 6 months, and 12 months after implantation, respectively. After retrieving the implanted valved conduits, histopathologic examination with Hematoxylin-Eosin, Masson' trichrome staining and immunohistochemical staining was performed. Result: Among the six goats, which had been implanted with acellularized pulmonary valved conduits, five survived the expected time period. Echocardiographic examinations for pulmonary valves revealed good function except mild regurgitation and stenosis. Microscopic analysis of the leaflets showed progressive cellular in-growth, composed of fibroblasts, myofibroblasts, and endothelial cells, into the acellularized leaflets over time. Severe inflammatory respon-se was detected in early phase, though it gradually decreased afterwards. The extracellular matrices were regenerated by repopulated cells on the recellularized portion of the acellularized leaflet. Conclusion: The acellularized xenogenic pulmonary valved conuits were repopulated with fibroblasts, myofibroblasts, and endothelial cells of the recipient and extracellullar matrices were regenerated by repopulted cells 12 months after the implantation. The functional integrity of pulmonary valves was well preserved. This study showed that the acellularized porcine xenogenic valved conduits could be used as an ideal valve prosthesis with long term durability.

Effects of 3-dimensional Co-culture of Human Endometrial Cells Decidualized with Progesterone and TGF-${\beta}1$ on the Development of Mouse 2-cell Embryos In Vitro (Progesterone과 TGF-${\beta}1$에 의해 탈락막화가 유도된 인간 자궁내막세포의 삼차원 공배양이 2-세포기 생쥐배아의 체외발달에 미치는 영향)

  • Kwon, Wook-Hyun;Kim, Hwi-Gon;Lee, Dong-Hyung;Ko, Kyung-Rae;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.35 no.1
    • /
    • pp.49-60
    • /
    • 2008
  • Objective: This study was carried out to investigate the effects of 3-dimensional co-culture of human endometrial cells decidualized with progesterone and TGF-${\beta}1$ on the development of 2-cell mouse embryos. Methods: Stromal and epithelial cells isolated from human endometrial tissue were immunostained for cytokeratin and vimentin. Expression of TGF-${\beta}1$, its receptor-1, -2, integrin-${\beta}3$ and prolactin in mono or co-culture according to three different hormone conditions was investigated by RT-PCR. Differential staining was used to investigate the number of ICM and trophectoderm of hatched mouse blastocysts in different three conditions. Results: The immunohistochemical study was positive for cytokeratin or vimentin and confirmed that epithelial and stromal cells were isolated from endometrial tissue successfully. In co-culture, TGF-${\beta}1$, its receptor-1, integrin-${\beta}3$ and prolactin except TGF-${\beta}1$-r2 were expressed in progesterone dominant condition. The hatching and attaching rate were higher in the co-culture with decidualized cells (p<0.05). However, we observed that lots of the incomplete hatched blactocysts attached on non-decidualized cells. The ICM number of hatched mouse blastocysts was higher in co-culture with decidualized and non decidualized cells than media only culture (p<0.05). The trophectoderm number of hatched blastocyst was higher in the co-culture with decidualized cells than non-decidualized cells or media only culture (p<0.05). Conclusion: The administration of progesterone, estrogen and TGF-$\beta$ could induce decidualization of stromal and epithelial cells isolated from human endometrial tissue using 3-dimensional co-culture, and the decidualization of human endometrial cells could increase the hatching and attaching rate of 2-cell mouse embryos.

Radiation-Induced Apoptosis of Lymphocytes in Peripheral Blood (말초혈액 내 림프구의 방사선에 의한 아포프토시스)

  • Oh, Yoon-Kyeong;Lee, Tae-Bum;Nam, Taek-Keun;Kee, Keun-Hong;Choi, Cheol-Hee
    • Radiation Oncology Journal
    • /
    • v.21 no.1
    • /
    • pp.75-81
    • /
    • 2003
  • Purpose : This study quantitatively evaluated the apoptosis In human peripheral blood lymphocytes using flow cytometry, and investigated the possibility of using this method, with a small amount of blood, and the time and dose dependence of radiation-induced apoptosls. Materials and Methods : Peripheral blood lymphocyes were isolated from the heparinized venous blood of 11 healthy volunteers, 8 men and 3 women, with each 10 ml of blood being divided Into IS samples. The blood lymphocytes were Irradiated using a linear accelerator at a dose rate of 2.4 Gy/min, to deliver doses of 0.5, 1, 2 and S Gy. The control samples, and Irradiated cells, were maintained in culture medium for 24, 48 and 72 hours fellowing the Irradiation. The number of apoptotic cells after the in vitro X-irradiation was measured by flow cytometry after Incubation periods of 24, 48 and 72 hours. We also observed the apoptotic cells using a DNA fragmentation assay and electron microscopy. Results : The rate oi spontaneous apoptosis increased in relation to the time interval following irradiation (1.761 ${\pm}$0.161, 3.563${\pm}$0.554, 11.098${\pm}$2.849, at 24, 48, and 72 hours). The apoptotli cells also increased In the samples irradiated with 0.5, 1, 2 and 5 Gy, In a radiation dose and time interval after Irradiation manner, with the apoptosls being too great at 72 hours after Irradiation. The dose-response curves were characterized by an Initial steep Increase In the number of apoptotic cells for Irradiation doses below 2 Gy, with a flattening of the curves as the dose approached towards 5 Gy. Conclusion :The flow cytometric assay technique yielded adequate data, and required less than 1 mL of blood. The time and dose dependence of the radiation-induced apoptosis, was also shown. It is suggested that the adequate time Interval required for the evaluation of apoptosis would be 24 to 48 hours after blood sampling.

Investigation on Antioxidant Activity in Plant resources (식물자원의 항산화활성 탐색)

  • Lee, Seung-Eun;Sung, Jung-Sook;Jang, In-Bok;Kim, Geum-Sook;Ahn, Tae-Jin;Han, Hee-Sun;Kim, Ji-Eun;Kim, Young-Ock;Park, Chung-Berm;Cha, Sun-Woo;Ahn, Young-Sup;Park, Ho-Ki;Bang, Jin-Ki;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
    • /
    • v.16 no.5
    • /
    • pp.356-370
    • /
    • 2008
  • This study was conducted for screening on antioxidant activity of 429 plants and selecting new potential antioxidant candidates. In vitro test models such as scavenging activity on DPPH radical and inhibitory activity on linoleic acid oxidation were used in the preliminary study. Flower of Sanguisorba officinalis, flower of Sedum kamtschaticum, flower of Rumex obtusifolius, and root of Sedum kamtschaticum showed very effective antioxidant activity on DPPH radical and linoleic acid oxidation. Those plants showed 8.1, 9.4, 9.9, $11{\mu}g/ml$ in DPPH radical scavenging activity as $SC_{50}$ and did 80.4, 80.1, 84.5, 88.0% in inhibition activity on linoleic acid oxidation, respectively. Root of Sedum middendorfianum M. showed positive effects in superoxide radical scavenging activity ($38.4{\mu}g/ml$) and inhibitory effect on $CuSO_4$-induced LDL oxidation (53.8% at final concentration of $1{\mu}g/ml$). Gleditsia japonica Mig. showed high antioxidant activity on LDL oxidation as 71.6% at final concentration of $1{\mu}g/ml$ and total phenol content of 958.5 mg% as tannic acid equivalent. In conclusion, we think that these plants having potent antioxidant activity might be studied further and could be used as new resources for many purposes including healthy food, functional cosmetics and drug development etc.

Antigen analysis of Toxoplasma gondii Iysate and excretory-secretory materials by enzyme-linked immunoelectrotransfer blot (EITB) (효소면역 전기영동이적법에 의한 톡소포자충 용해물 및 분비 항원의 분석)

  • 안명희;손혁진
    • Parasites, Hosts and Diseases
    • /
    • v.32 no.4
    • /
    • pp.249-258
    • /
    • 1994
  • Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

  • PDF

Anti-inflammatory Effect of Lactuca sativa L. Extract in Human Umbilical Vein Endothelial Cells and Improvement of Lipid Levels in Mice Fed a High-fat Diet (상추 추출물(Lactuca sativa L.)의 혈관내피세포에서 항염증 작용과 고지방 식이 생쥐에서 혈중 지질농도 개선에 미치는 영향)

  • Hwang-Bo, Jeon;Jang, Kyung Ok;Chung, Hayoung;Park, Jong-Hwa;Lee, Tae Hoon;Kim, Jiyoung;Chung, In Sik
    • The Korean Journal of Food And Nutrition
    • /
    • v.29 no.6
    • /
    • pp.998-1007
    • /
    • 2016
  • The objective of this study was to investigate the effects of a lettuce (Lactuca sativa L.) extract on the inflammation of human umbilical vein endothelial cell (HUVEC) and blood lipid improvement in hypercholesterolemic mice fed a high cholesterol diet. The lettuce extract (100% ethanol extract) inhibited the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVEC treated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$). The lettuce extract suppressed the adhesion of THP-1 to TNF-${\alpha}$-treated HUVEC. The lettuce extract decreased the TNF-${\alpha}$-stimulated production of proinflammatory cytokine interleukin-6, interleukin-8 and chemokine monocyte chemotactic protein 1. In hypercholesterolemic mice, the lettuce extract reduced serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol level, while the lettuce extract elevated high-density lipoprotein-cholesterol level, resulting in the decrease of atherogenic index and cardiac risk factor level. These results suggested that lettuce extract can be an useful resource to show an anti-inflammatory effect and improve lipid metabolism.