• Title/Summary/Keyword: in vitro effect

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Effect of Thiol Compounds and Antioxidants on In Vitro Development and Intracellular Glutathione Concentrations of Bovine Embryos Derived from In Vitro Matured and In Vitro Fertilized I. Effect of $\beta$-Mercaptoethanol and Cysteamine on Development and Intracellular Glutathione Concentrations of Bovine IVM/IVF Embryos (Thiol 화합물과 황산화제 첨가배양이 소 체외수정란의 체외발육과 세포내 Glutathione 농도 변화에 미치는 효과 I. $\beta$-Mercaptoethanol과 Cysteamine 첨가가 소 체외수정란의 체외발육과 세포내 Glutathione 농도 변화에 미치는 영향)

  • 양부근;박동헌;정희태;박춘근;김종복;김정익
    • Korean Journal of Animal Reproduction
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    • v.21 no.4
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    • pp.335-343
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    • 1997
  • The effect of thiol compounds on development and intracellular glutathione(GSH) concentrations of bovine embryos produced by in vitro maturation and in vitro fertilization(IVM/IVF) was examined in CRlaa medium with or without $\beta$-mercaptoethanol(0, 10, 25 and 50$\mu$MME) and cysteamine(0, 25, 50 and 75 $\mu$M). Numbers of cells comprising blastocysts were also counted using double fluorescence stain and the total glutathione levels(oxidized and reduced form) of morula and blastocyst embryos were than measured by an enzymatic method. Following routine IVM/IVF procedures oocytes and zygotes were cultured for 40 to 44h in CRlaa medium. Then 2 to 8-cell embyos had cumulus cell removed and were allotted randomly to the experimental medium. In Experiment 1, the proportion of embryos developing to and beyond morulae stages in 0, 10, 25 and 50 $\mu$M $\beta$-ME was 42.9%, 50.0%, 53.7% and 65.6%, respectively. Fifty $\mu$M $\beta$-ME group was significantly higher than those of any other groups (P<0.05). In Experiment 2, the percentages of embryos developed beyond morulae stages in 0, 25, 50 and 75 $\mu$M cysteamine was 42.9%, 40.4%, 60.0% and 59.2%, respectively. Fifty and 75$\mu$M cysteamine groups were significantly higher than in 0 and 25 $\mu$M cysteamine groups, but all of culture medium containing cysteamine(52.6%) was not significantly difference in control group(42.9%). In Experiment 3, the intracellular GSH concentrations of morulae and blastocyst embryos in 0 and 50 $\mu$M $\beta$-ME was 42.4 pM and 44.9 pM, 49.5 pM and 67.8 pM, respectively. Morulae embryos were not difference, but blastocyst embryos were significantly difference between treatments(P<0.05). In Experiment 4, the intracellular GSH concentrations of morulae in CRlaa with or without cysteamine were 39.8 pM and 45.6 pM, and blastocysts were 59.3 pM and 66.8 pM, respectively. Cell numbers of blastocysts were similar to in all experimental groups. These experiments indicate that thiol compounds can increase the proportion of embryos that developing to and beyond morulae stage and the intracellular GSH concentrations.

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IN VITRO EFFECT OF PANAX GINSENG ON PHYTOHAEMAGGLUTININ-INDUCED LYMPHOCYTE TRANSFORMATION AND ITS IN VIVO EFFECT (식물성 혈구 응집소(Phytohaemagglutinin)로 유도한 림파구 변형에 관한 인삼의 in vitro 및 in vivo효과)

  • Chong S.K.F.;Brown H.A.;Rimmer E.;Oberholzer V.;Hindocha P.;Walker-Smith J .A.;Carruthers L.
    • Proceedings of the Ginseng society Conference
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    • 1984.09a
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    • pp.89-95
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    • 1984
  • The effect of hydrocortisone or Panax ginseng, and/or a combination of hydrocortisone and Panax ginseng on phytohaemagglutinin (PHA-P)-induced transformation of peripheral blood lymphocytes were studied in 4 normal healthy adult volunteers. Increasing concentrations of Panax ginseng 0.16-1.60${\mu}g$/ml caused a doserelated inhibition of PHA-P transformation of lymphocytes. A combination of 500${\mu}g$/ml hydrocortisone and 0.80${\mu}g$/ml Panax ginseng produced a greater suppression of PHA-P stimulation than either drug used alone. This suggests that Panax ginseng has a steroid-like effect in vitro, and may have a potentiating effect with hydrocortisone on T-cell mediated immunity. The in vivo effects of Panax ginseng were further studied in 6 healthy adult volunteers. PHA-P transformation studies were performed before and after Panax ginseng capsules were taken for 2 weeks and at a higher dosage at 4 weeks. Our results showed that in 3 subjects, there was significant inhibition of PHA-P transformation at 4 weeks.

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Anti-inflammatory Effects of Goihwa-san Water Extract via NF-κB Inhibition (괴화산(槐花散)의 NF-κB 기전을 통한 항염증 효과 연구)

  • Hyun Hee Cho;Ji Young Choi;Min Hwangbo;Seon Young Jee
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.36 no.1
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    • pp.21-39
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    • 2023
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Goihwa-san water extract(GHS) in vitro & in vivo. Methods : In vitro, we evaluated the anti-inflammatory effect of GHS by comparing the Raw 264.7 cells with 10, 30, 100, 300㎍/㎖ of GHS for 1 hour before Lipopolysaccharide(LPS) to the single LPS treated group. We examined the relative cell viability by MTT assay and the relative level of LPS, Loxoribine(LOX), Peptidoglycan(PGN), Flagellin(FLA)-induced NO production by using Griess reagent and measured relative iNOS protein level and COX-2 protein level by using western blot and Image analyzing system. We measured the production of TNF-α, IL-1β, and IL-6 by each ELISA kits and then measured the relative levels of IκBα, p-IκBα in whole-cell lysate fraction and NF-κB in nuclear fraction by using western blot and Image analyzing system. In vivo, we induced the paw edema by subcutaneous injection of 100㎕/rat CA and measured the swelling volume of paw by using a plethysmometer and then measured the relative iNOS protein level by using western blot. Results : As a result, in vitro, LPS, PGN-induced NO production was significantly inhibited by pretreatment with GHS. GHS reduced LPS, PGN-induced iNOS expression, PGN-induced COX-2 expression and LPS-induced production of cytokine(TNF-α, IL-1β, IL-6). Expression of IκBα was increased by pretreatment with GHS 100㎍/㎖. And the expression of p-IκBα and NF-κB were decreased by pretreatment with GHS 100㎍/㎖. In vivo, CA-induced inflammation rat model was used for the evaluation of the anti-inflammatory effect of GHS. 0.3 or 1.0g/kg of GHS significantly reduced the increases of paw swelling and iNOS expression in paw tissues. Conclusions : These results show that GHS can decrease inflammatory response via inhibition of the NF-κB pathway in vitro. And in vivo, the anti-inflammatory effect suggest the clinical basis of GHS for the treatment of inflammatory diseases.

Comparison of in vitro ruminal fermentation incubated with different levels of Korean corn grains with total mixed ration as a basal

  • Hamid, Muhammad Mahboob Ali;Park, Ha Young;Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.419-427
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    • 2018
  • The present study was conducted to investigate the effect of different levels of Korean corn grain on in vitro ruminal fermentation with total mixed ration (TMR) as a basal feed. Three ruminal cannulated Holstein steers (Body Weight $479{\pm}33.0kg$) were used as rumen fluid donors. Treatments for in vitro fermentation were TMR only (control, 3.0 g), TMR substituted partially with high level (HC, TMR 1.5 and corn 1.5 g), and with low level of Korean corn grain (LC, TMR 2.25 and corn 0.75 g), respectively. To measure in vitro ruminal pH, gas production, ammonia N and volatile fatty acids (VFA), the in vitro fermentation incubation was triplicated at $39^{\circ}C$, 120 rpm for 0, 1, 3, 6, 12, 24 and 48 h, respectively. Mean ruminal pH was significantly lower (p < 0.05) for HC than control. Changes in rumen pH was rather similar between the groups till 6 h after incubation, but the lowest pH for HC (pH 5.10) appeared at 48 h compared with control and LC. Total gas production was tended (p < 0.09) to be higher and ammonia N was significantly lower (p < 0.05) for HC than control and LC. Total VFA was higher (p < 0.05) for HC and LC than control but no differences appeared between HC and LC. Overall, the present data indicate that feeding different levels of Korean domestic corn grain may lead to high and sustainable starch degradation in the rumen.

Effect of ${\beta}$-glucan Originated from Aureobasidium on the Dermal Wound Repair in Vitro Model (생체외 모델에서 아우레오바지디움 유래 베타 글루칸이 피부창상 치유에 미치는 영향)

  • Lee, Jung-Woo;Kwon, Young-Sam;Jang, Kwang-Ho
    • Journal of Veterinary Clinics
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    • v.30 no.6
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    • pp.403-408
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    • 2013
  • The objective of the present study is to detect the effect of ${\beta}$-glucan originated from Aureobasidium on the proliferation and collagen production in human dermal fibroblast cells with wound repopulation in vitro. The proliferative effects were assessed using a MTT assay as well as cell counts at 24 and 48 hr after treatment. Hydroxyproline was measured as an index of procollagen production with reverse-phase high pressure liquid chromatography. Oncostatin M was used as a reference agent. In glucagon treated group, dose-dependent and significant increase of optical density or fibroblast cell numbers was demonstrated, when compared with those of control from 0.1 mg/ml concentration. In addition, the numbers of cells which had migrated into the wound defects were more significantly and dose-dependently increased than those of non-treated control. However, no meaningful effects on the procollagen production were observed.

Gametophyte Propagation and Sporophyte Formation of Asian Chain Fern [Woodwardia japonica (L. f.) Sm.] Under Various Medium Conditions In Vitro and Ex Vitro

  • Cho, Ju Sung;Jang, Bo Kook;Park, Kyungtae;Lee, Ha Min;Lee, Cheol Hee
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.735-742
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    • 2019
  • This study investigated a suitable method that could be applied for Asian chain fern [Woodwardia japonica (L. f.) Sm.] to propagate gametophytes and promote sporophyte formation. The gametophytes used in all experiments were obtained from germinated spores in vitro and were subcultured at 8-week intervals. The most appropriate media for gametophyte propagation was identified by culturing 300 mg of gametophyte in Murashige and Skoog (MS) basal medium (1/8, 1/4, 1/2, 1, 2), and Knop medium for 8 weeks. As a result, fresh weight of the gametophyte was increased by 56.7-fold on MS medium. Moreover, antheridium formation as well as gametophyte growth was improved on MS medium, especially. To improve the sporophyte formation ex vitro, 1.0 g of gametophyte was ground with distilled water and spread on eight combinations onto four different culture mediums, such as bed soil, peat moss, perlite and decomposed granite. Then generation and growth of sporophytes were investigated after cultivation for 10 weeks. As a result of this experiment, peat moss had a promotive effect of sporophyte formation at single-use and mixed culture soils. In particular, a mixture of bed soil, peat moss and perlite in a 1:1:1 ratio (v/v/v) led to the accelerated formation (782.5 ea/pot) and the frond growth of sporophytes. This included increases in length and width of fronds. However, promotive effect of gametophyte growth and sporophyte formation was not found at single-use and treatment with high ratio of bed soil.

The Effect of Pyroligneous Liquor and Coconut Water on Plantlet Multiplication and in Vitro Flowering of Dendrobium moniliforme (목초액 및 코코넛액이 석곡(Dendrobium moniliforme)의 유묘 증식과 기내 개화에 미치는 영향)

  • Jee, Sun-Ok;Cho, Dong-Hoon
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.739-742
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    • 2005
  • This experiment was carried out to clarify the effect of pyroligneous liquor and coconut water on plantlet multiplication and in vitro flowering of Dendrobium moniliforme. Plantlet growth and multiplication was good in 1.0 ml/L pyroligneous liquor treatment which was added to the basal media of 3 g/L hyponex and 4 g/L peptone ($H_{3} P_{4}$) containing 0.1 mg/L NAA and 1.0 mg/L kinetin. In the treatment of 30 ml/L coconut water showed good results on plantlet growth and multiplication. In vitro flowering showed highest rate in the treatment of 1.0 ml/L pyroligneous liquor and 30 ml/L coconut water which were added to the basal media of $H_{3} P_{4}$ containing 0.1 mg/L NAA and 1.0 mg/L kinetin.

Effect of Lactic Acid Bacteria Treatment on Nutritive Value and In Vitro Ruminal Fermentation of Italian Ryegrass (Lolium multiflorum L.) Silage

  • Lee, Kihwan;Marbun, Tabita Dameria;Kim, Suyeon;Song, Jaeyong;Kwon, Chan Ho;Yoon, Duhak;Kang, Jungsun;Lee, Chanho;Cho, Sangbuem;Kim, Eun Joong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.3
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    • pp.182-189
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    • 2020
  • This study was conducted to evaluate the effect of lactic acid bacteria (LAB) inoculation to domestically-cultivated Italian ryegrass (IRG) on silage fermentation and in vitro ruminal fermentation. There were six treatments based on the LAB inoculants: 1) no addition of LAB (negative control: NC), additions of 2) commercially-available LAB (positive control: PC), 3) Lactobacillus plantarum (LPL), 4) L. paracasei (LPA), 5) L. acidophilus (LA), and 6) L. pentosus (LPT). All treatments were inoculated at a concentration of 106 CFU/g and ensiled for 3, 7, 21, and 42 days in triplicate and analyzed for nutritive values when ensiling was terminated. Day 42 silage from all treatments were also examined for in vitro ruminal fermentation. After 42 days, LAB-inoculated silages had higher (P<0.05) lactic acid concentration compared to the NC. In terms of nutritive values, the silages treated with LPA, LA, and LPT showed higher (P<0.05) crude protein and lower (P<0.05) neutral detergent fiber and acid detergent fiber content compared to the rest of the treatment. In vitro ruminal dry matter degradability was not affected by LAB addition. However, LAB-treated IRG had shown higher (P<0.05) ammonia-N compared with that of the NC. LPA had shown the highest (P<0.05) volatile fatty acid concentration among the LAB examined. In conclusion, the addition of a single strain of LAB appeared to produce a quality IRG silage compared with the NC and the PC. Among the strains examined, LPA seemed to be superior to the others.

Effect of Pantothenic Acid, Myo-Inositol, and Folic Acid on In Vitro Development of Parthenogenetic Pig Embryos (Pantothenic Acid, Myo-Inositol 및 Folic Acid가 돼지 단위 발생 배아의 체외발육에 미치는 영향)

  • You, Jin-Young;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.25 no.1
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    • pp.1-7
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    • 2010
  • The objective of this study was to examine the effect of vitamin B (pantothenic acid, folic acid, and myo-inositol) that was supplemented to embryo culture medium on in vitro development of parthenogenetically activated (PA) pig embryos. Cumulus-oocyte complexes derived from slaughtered ovaries were matured in TCM-199 supplemented with porcine follicular fluid, cysteine, pyruvate, EGF, insulin, and hormones (hCG and eCG) for the first 22 h and then further cultured in hormone-free medium for an additional 22 h. After maturation culture, metaphase II oocytes that extruded 1st polar body were electrically activated and treated with $5.0\;{\mu}g/ml$ cytochalasin B for 4 h. Then, PA embryos were cultured for 7 days in a modified NCSU-23 that was supplemented with pantothenic acid, myo-inositol, or folic acid at different concentrations ($3{\sim}300\;{\mu}M$) according to the experimental design. Myo-inositol added to culture medium did not show any beneficial or inhibitory effects on embryo cleavage and blastocyst formation. However, $300\;{\mu}M$ pantothenic acid significantly inhibited blastocyst formation compared to control (no addition) (24% vs. 36%, p<0.05). Folic acid ($300\;{\mu}M$) significantly (p<0.05) increased blastocyst formation (56%) compared to control (41%). Our results demonstrated that in vitro development of PA embryos was significantly influenced by vitamin B and addition of $300\;{\mu}M$ folic acid to culture medium improved in vitro development of pig PA embryos.

Colony Count with Mixed Culture of Enteric Bacteria by in vitro Quantitative Method (장내세균의 시간차 혼합배양이 보여주는 균수측정의 비교)

  • 황선철;전보성
    • Korean Journal of Microbiology
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    • v.11 no.4
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    • pp.175-180
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    • 1973
  • This study was attempted to see more clear relationships among the enterobacteria, especially between the intestinal normal flora and pathogenic bacteria. It has been known that some intestinal normal flora produce the bactrial metabolites that are harmful to other enteric bacteria. One of the metabolites is known as colicin, the protein fraction, which possesses certain degree of inhibitory effect against other bacterial growth fraction, whih possesses certain degree of inhibitory effect against other bacterial growth. As a preliminary study for a colicin purification, the antagonistic effect of E, coli to groups of Salmonella and Shigella has been studied by means of in vitro quantitative culture method. 1. E.coli showed definite inhibitory effects aganist both Salmonella and Shigella groups in the mixture of two organisms. 2. The inhibitory effects of E.coli in the E.coli-Salmonella and the E.coli-Shigella mixture occurred from 4 hours incubation following the inoculation. 3. Even the complete inhibition of pathogenic enteric bacterial growth was noticed in the E.coli-Salmonella mixture at overnight incubation. 4. Among the diluted mixtures, 1:100, 1:1,000, and 1:10,000, survival rate of pathogenic enteric bacteria in the mixtures with E.coli showed least affected at the 1:1,000 dilution. 5. It was found that the antagonistic effect aganist groups of Salmonella-shigella was depending upon the groups of the genera.

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