• Title/Summary/Keyword: in vivo experiment

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The Immunomodulating Effects of the Supplementation of Paeonia Japonica Extracts in Mice

  • Kim, Jin;Kim, Hyun-Sook
    • Nutritional Sciences
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    • v.5 no.2
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    • pp.60-67
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    • 2002
  • Paeonia japonica var. pilosa $N_{AKAI}$, (PJ; Baek-Jak-Yak) is a medicinal plant which has been widely used as a component or blood-building decoctions. This study was performed to investigate the immunomodulative effects of PJ in mice, using in vitro and in vivo experiments. The immunomodulative effects were studied in vitro by determining the proliferation or mice splenocytes and the production of three kinds of cytokines (IL-1$\beta$, IL-6, TNF-$\alpha$) by mire peritoneal macrophages which were cultured with sequential fractions of PJ methanol extract (methanol, hexane, chloroform, ethylacetate, butanol and water). In an in vivo experiment using mice, different concentrations of PJ water extract were orally administrated every other day for two weeks. The production of cytokines (IL-1$\beta$, IL-6, TNF-$\alpha$) secreted by activated macrophages, and the proliferation of mice splenocytes, were used as indices for immunocompetence. In vitro supplementation using a hexane fraction of PJ in the range of 1 to 100 $\mu$ g/ml enhanced splenocyte proliferation by 1.8 to 12%, and by 10-15% using an aqueous fraction, compared to the control. IL-l$\beta$ production was significantly increased with the supplementation of butanol, hexane and water extracts of PJ Higher levels of IL-6 production were detected with supplementation of chloroform or water extracts. However, there were no significant differences in the production of TNF-$\alpha$ among the treated groups and the control. From the in vivo study, the highest proliferation of splenocytes was seen in the mice orally administrated with the PJ water extract at the concentration of 500 mg/kg body weight. In the case of cytosine production, IL-1-$\beta$, IL-6, and TNF-$\alpha$ released by activated peritoneal macrophages were augmented by the oral administration of a PJ water extract. These results indicate that Pl may enhance the immune function by regulating splenocyte proliferation and cytokine production capacity in mice.

Effect of Aralia Cordata Pharmacopuncture on Cartilage Protection and Apoptosis Inhibition In Vitro and in Collagenased-induced Arthritis Rabbit Model

  • Park, Dong-Suk;Baek, Yong-Hyeon
    • The Journal of Korean Medicine
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    • v.28 no.4
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    • pp.114-123
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    • 2007
  • Osteoarthritis is characterized by cartilage degradation and chondrocytes death. Chondrocyte death is induced by the apotosis through special mechanisms including the activation of caspase-3. On the basis of this background, this study was designed to examine the cartilage protective and anti-apototic effects of Aralia Cordata in in vtro and in collagenase-induced arthritis rabbit model. To conduct in vitro study, chondrocytes culturedfrom rabbit knee joint were treated by 5 ng/ml IL-1a.For in vivo experiment, collagenase-induced arthritis (CIA) rabbit model was made via intraarticular injection with 0.25 ml of collagenase solution. Aralia cordata pharmacopuncture (ACP) was administrated on bilateral Dokbi acupoint (ST35) of rabbits at a dosage of 150 ${\mu}g/kg$ once a day for 28 days after the initiation of the CIA induction. In the study by using CIA rabbit model in vivo, ACP showed the inhibition of cartilage degradation in histological analysis. Aralia cordata also showed anti-apoptotic effect both in vitro and in vivo study. In chondrocytes treated by IL-1a, Aralia cordata inhibited caspase-3 activity and enhanced the proliferation of IL-1a-induced dedifferentiated chondrocytes. ACP showed the inhibition effect on the caspase-3 expression and activity from CIA rabbit model. This study indicates that ACP inhibits the cartilage destruction and the chondrocyte apotosis through downregulation of caspase-3 activity. These data suggest that ACP has a beneficial effect on preventing articular cartilage destruction in osteoarthrtis.

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Control of the Intraocular Pressure by Using the Micro Valve Actuator for a Glaucoma Implant

  • Byunghoon Bae;Park, Junhyun;Park, Kyihwan;Lee, Jonghyun;Lee, Yeon;Hongseok Kee;Kim, Seonho;Taeseok Sim;Kim, Yongkweon
    • 제어로봇시스템학회:학술대회논문집
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    • 2002.10a
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    • pp.74.6-74
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    • 2002
  • $\textbullet$ A new implant capable of controlling IOP actively and coping with personal difference among patients $\textbullet$ The valve actuator of the active implant is proposed and fabricated. $\textbullet$ In vitro experiments are carried out extensively with and without a rabbit. $\textbullet$ The experiments are conducted by using the open and closed loop pressure control. $\textbullet$ The experiment with the rabbit is similar as a in vivo experiment, $\textbullet$ since the compliance and the aqueous humor of the rabbit's eye are included. $\textbullet$ The experimental results verify the possibility of the valve actuator for a glaucoma implant.

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Cytochalasin E Production by Rosellinia necatrix and Its Pathogenicity on Apple (사과흰날개무늬병균의 Cytochalasin E 독소 생산과 병원성)

  • Lee, Dong-Hyuk;Choi, Kyung-Hee;Uhm, Jae-Youl
    • Research in Plant Disease
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    • v.15 no.1
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    • pp.46-50
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    • 2009
  • Cytochalasin E (CE) is a secondary metabolite secreted by Rosellinia necatrix, caused by white root rot, and has toxicity to apple as a toxin during disease progress. This study was conducted to demonstrate the relationship between the production of CE and its pathogenicity. CE producing isolates and non-producing isolates of R. nectatrix were isolated from the mycerial mat of diseased roots and was detected on that using a TLC and HPLC analysis and in vivo pathogenicity test. CE non-producing isolates were not pathogenic to apple roots and not detected CE by TLC and HPLC analysis. It was shown that the production of CE was related to the pathogenicity of R. nectatrix.

Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng

  • Song, Jeong Young;Seo, Mun Won;Kim, Sun Ick;Nam, Myeong Hyeon;Lim, Hyoun Sub;Kim, Hong Gi
    • Mycobiology
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    • v.42 no.2
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    • pp.174-180
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    • 2014
  • We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

In Vivo Hair Growth Promotion Effects of Ultra-High Molecular Weight Poly-γ-Glutamic Acid from Bacillus subtilis (Chungkookjang)

  • Choi, Jae-Chul;Uyama, Hiroshi;Lee, Chul-Hoon;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.407-412
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    • 2015
  • We investigated the effect of ultra-high molecular weight poly-γ-glutamic acid (UHMW γ-PGA) on hair loss in vitro and in vivo. 5-Alpha reductase is an enzyme that metabolizes the male hormone testosterone into dihydrotestosterone. By performing an in vitro experiment to analyze the inhibitory effects of UHMW γ-PGA on 5-alpha reductase activity, we determined that UHMW γ-PGA did in fact inhibit 5-alpha reductase activity, indicating the use of UHMW γ-PGA as a potential 5-alpha reductase inhibitor in the treatment of men with androgenetic alopecia. To evaluate the promotion of hair growth in vivo, we topically applied UHMW γ-PGA and minoxidil on the shaved dorsal skin of telogenic C57BL/6 mice for 4 weeks. At 4 weeks, the groups treated with UHMW γ-PGA showed hair growth on more than 50% of the shaved skin, whereas the control group showed less hair growth. To investigate the progression of hair follicles in the hair cycle, hematoxylin and eosin staining was performed. Histological observations revealed that the appearance of hair follicles was earlier in the UHMW γ-PGA-treated group than in the control group. The number of hair follicles on the relative area of shaved skin in the UHMW γ-PGA-treated group was higher than that observed on the shaved skin in the control group. These results indicate that UHMW γ-PGA can promote hair growth by effectively inducing the anagen phase in telogenic C57BL/6 mice.

Morphological Characteristics of Pig Blastocysts Produced by Somatic Cell Nuclear Transfer

  • Y.M. Han;D.B. Koo;Park, Y.H.;Park, J.S.;Kim, H.N.;Y.K. Kang;W.K. Chang;Lee, K.K.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.68-68
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    • 2001
  • Blastocyst formation, consisting of the inner cell mass (ICM) and trophectoderm (TE), is the first differentiation process during embryonic development in mammals. It has been hypothesized that the proportion of ICM to TE in the blastocyst may be crucial for subsequent developmental competence of early embryos, which it may be expressed as a sensitive indicator for evaluating in vitro systems. In this study ICM/total cell ratio of nuclear transfer (NT) embryos was compared with IVF-derived and in vivo embryos. Somatic cell nuclei obtained from a fetus at Day 40 of gestation were transferred into the enucleated oocyte and then cultured in NCSU 23 medium for 6 days as previously described (Koo et al., Biol. Reprod. 2000; 63:986-992). ICM and TE cells of blastocysts were determined by using a differential staining method (Han et al., Biol. Reprod. 1999; 60:1110-1113). Development rate (9.8$\pm$2.5%, 23/225) to the blastocyst stage of NT embryos was lower than IVF embryos (23.8$\pm$2.7%, 53/223). Thus, a difference was detected in the in vitro developmental rate to blastocyst stage between NT and IVF-derived embryos (P<0.05). In the next experiment, we investigated ICM and TE nuclei to assess the quality of blastocysts that produced by NT, IVF and in vivo, respectively. NT blastocysts (27.6$\pm$8.3) showed a smaller total cell number than IVF-derived (42.6$\pm$17.4) and in vivo embryos (283.9$\pm$103.5) (P<0.05). Ratios of ICM/total cells in NT, IVF and in vivo blastocysts were 15.1$\pm$ 18.6% (n=56), 12.3$\pm$9.2% (n=57) and 30.4$\pm$6.8% (n=40), respectively. Individual blastocysts for the ratio of ICM/total cells were assigned to 3 groups (I; <20%, II; 20 to 40% and III;>40%). As the results, most in vivo blastocysts (97.5%, 39/40) were distributed into group II while most NT (78.6%, 44/56) and IVF-derived blastocysts (82.5%, 47/57) were allocated to group I. Thus, our data show that NT or IVF-derived embryos have aberrant morphology during early development in vitro systems, suggesting that these anomalies may result in developmental failures of the NT embryos to term.

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Functional Evaluation of the Rockbream (Oplegnathus fasciatus) Beta-actin Promoter as a Candidate Regulatory Element for DNA Vaccination

  • Kosuke, Zenke;Lee, Sang-Yoon;Kim, Ki-Hong;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.12 no.2
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    • pp.98-103
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    • 2009
  • The potential utility of the rockbream (Oplegnathus fasciatus) $\beta$-actin 5'-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream $\beta$-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream $\beta$-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the $\beta$-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream $\beta$-actin promoter could be used as a potential regulator for DNA vaccination in this species.

Influence of (-)-Hydroxycitrate on food Intake, Body Weight and Lipogenesis in Rats ((-)-Hydroxycitrate의 식이 투여가 흰쥐의 식이 섭취량, 체중, 지방대사 및 합성에 미치는 영향)

  • 김상배
    • Journal of Nutrition and Health
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    • v.30 no.2
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    • pp.123-131
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    • 1997
  • The influence of (-0-Hydroxycitrate(HCA), shown to be a competitive inhibitor of adenosine 5-triphosphate(ATP) citrate lyase, on food intake and body weight, serum triglyceride and cholosterol level, in vivo rates of fatty acid and cholesterol synthesis, and fat cell number and size was investigated. 3 groups of female, 5 weeks old Sprague Dawley rats, 8 animals each, were ad libitum meal-fed or pair-fed(3 hours from 10 : 00 to 13 : 00) AIN based high glucose diet for a total period of 8 weeks. Providing normolipidemic rats orally with 400mg of HCA formula containing approximately 20mg of HCA 1 hour prior to daily feeding schedule significantly depressed in vivo hepatic rates of fatty acid and cholesterol synthesis in the liver and adipose tissue. Serum triglyceride and cholesterol levels were significantly reduced by HCA. At the end of treatment period, the rats administered with HCA resulted in a significantly reduction in body weight gain. The reduction in weights was attributable to a significant decrease in fat cell size with a smaller extent, but not significant, reduction in fat cell number. Rats receiving HCA demonstrated less food intake than the controls ; however, this decreased caloric intake was not fully responsible for the HCA induced depression of hepatic and adipocytic lipogenesis, since experiment using pair-fed cojntrol rats showed, less magnitude but similar results. Both a anorectic and an antilipogenic properties of HCA seem to be responsible for this weight reduction activity of HCA. The outcome of this study suggest that metabolic regulation may be a feasible approach to the control of obesity and hyperlipidemia.

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Therapeutic Effects of Bacteriophages Against Salmonella gallinarum Infection in Chickens

  • Hong, Sung Sik;Jeong, Jipseol;Lee, Jinju;Kim, Suk;Min, Wongi;Myung, Heejoon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1478-1483
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    • 2013
  • In this study the isolation and characterization of three bacteriophages (ST4, L13, and SG3) infecting Salmonella gallinarum were carried out. They were further tested for their in vivo efficacy in phage therapy. All three phages belong to the Siphoviridae family with isometric heads and non-contractile tails. They have a broad host range among serovars of Salmonella enterica. The burst sizes were observed to be 1670, 80, and 28 for ST4, L13, and SG3, respectively. The in vivo efficacy of the phages was tested in chickens. Layer chickens were challenged with S. gallinarum, whereas contact chickens were cohabited without direct challenge. Each bacteriophage was orally inoculated in the form of feed additives. Mortality was observed and S. gallinarum was periodically re-isolated from the livers, spleens, and cecums of the chickens. Bacterial re-isolation from the organs and mortality decreased significantly in both challenged and contact chickens treated with the bacteriophages compared with untreated chickens serving as the control. The three bacteriophages may be effective alternatives to antibiotics for the control of fowl typhoid disease in chickens.