• Title/Summary/Keyword: $In$ $vitro$ degradation

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Ralstonia pickettii Enhance the DDT Biodegradation by Pleurotus eryngii

  • Purnomo, Adi Setyo;Maulianawati, Diana;Kamei, Ichiro
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1424-1433
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    • 2019
  • DDT is a hydrophobic organic pollutant, which can be bio-accumulated in nature and have adverse consequences on the physical condition of humans and animals. This study investigated the relationship between the white-rot fungus Pleurotus eryngii and biosurfactant-producing bacterium Ralstonia pickettii associated with the degradation of DDT. The effects of R. pickettii on fungal development were examined using in vitro confrontation assay on a potato dextrose agar (PDA) medium. R. pickettii culture was added to the P. eryngii culture at 1, 3, 5, 7, and 10 ml ($1ml{\approx}1.44{\times}10^{13}CFU$). After 7 d incubation, about 43% of the initial DDT ($12.5{\mu}M$) was degraded by the P. eryngii culture only. The augmentation of 7 ml of R. pickettii culture revealed a more highly optimized synergism with DDT degradation being approximately 78% and the ratio of optimization 1.06. According to the confrontational assay, R. pickettii promoted the growth of P. eryngii towards the bacterial colony, with no direct contact between the bacterial cells and mycelium (0.71 cm/day). DDD (1,1-dichloro-2,2-bis(4-chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1-chloro-2,2-bis(4-chlorophenyl) ethylene) were identified as metabolic products, indicating that the R. pickettii could enhance the DDT biodegradation by P. eryngii.

Ginseng seed oil ameliorates hepatic lipid accumulation in vitro and in vivo

  • Kim, Go Woon;Jo, Hee Kyung;Chung, Sung Hyun
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.419-428
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    • 2018
  • Background: Despite the large number of studies on ginseng, pharmacological activities of ginseng seed oil (GSO) have not been established. GSO is rich in unsaturated fatty acids, mostly oleic and linoleic acids. Unsaturated fatty acids are known to exert a therapeutic effect in nonalcoholic fatty liver disease (NAFLD). In this study, we investigated the protective effect and underlying mechanisms of GSO against NAFLD using in vitro and in vivo models. Methods: In vitro lipid accumulation was induced by free fatty acid mixture in HepG2 cells and by 3 wk of high fat diet (HFD)-feeding in Sprague-Dawley rats prior to hepatocyte isolation. The effects of GSO against diet-induced hepatic steatosis were further examined in C57BL/6J mice fed a HFD for 12 wk. Results: Oil Red O staining and intracellular triglyceride levels showed marked accumulation of lipid droplets in both HepG2 cells and rat hepatocytes, and these were attenuated by GSO treatment. In HFD-fed mice, GSO improved HFD-induced dyslipidemia and hepatic insulin resistance. Increased hepatic lipid contents were observed in HFD-fed mice and it was lowered in GSO (500 mg/kg)-treated mice by 26.4% which was evident in histological analysis. Pathway analysis of hepatic global gene expression indicated that GSO increased the expression of genes associated with ${\beta}$-oxidation (Ppara, Ppargc1a, Sirt1, and Cpt1a) and decreased the expression of lipogenic genes (Srebf1 and Mlxipl), and these were confirmed with reverse transcription and quantitative polymerase-chain reaction. Conclusion: These findings suggest that GSO has a beneficial effect on NAFLD through the suppression of lipogenesis and stimulation of fatty acid degradation pathway.

Preparation and Bioevaluation of 177Lu-labelled Anti-CD44 for Radioimmunotherapy of Colon Cancer

  • Lee, SoYoung;Hong, YoungDon;Jung, SungHee;Choi, SunJu
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.187-192
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    • 2015
  • CD44 is a particular adhesion molecule and facilitates both cell-cell and cell-matrix interactions. In particular, splice variants of CD44 are particularly overexpressed in a large number of malignancies and carcinomas. In this study, the $^{177}Lu$-labelled CD44 targeting antibody was prepared and bioevaluated in vitro and in vivo. Anti-CD44 was immunoconjugated with the equivalent molar ratio of cysteine-based DTPA-NCS and radioimmunoconjugated with $^{177}Lu$ at room temperature within 15 minutes. The stability was tested in human serum. An in vitro study was carried out in HT-29 human colon cancer cell lines. For the biodistribution study $^{177}Lu$-labelled anti-CD44 was injected in xenograft mice. Anti-CD44 was immunoconjugated with cysteine-based DTPA-NCS and purified by a centricon filter system having a molecular cut-off of 50 kDa. Radioimmunoconjugation with $^{177}Lu$ was reacted for 15 min at room temperature. The radiolabeling yield was >99%, and it was stable in human serum without any fragmentation or degradation. The radioimmunoconjugate showed a high binding affinity on HT-29 colon cancer cell surfaces. In a biodistribution study, the tumor-to-blood ratio of the radioimmunoconjugate was 43 : 1 at 1 day post injection (p.i) in human colon cancer bearing mice. The anti-CD44 monoclonal antibody for the targeting of colon cancer was effectively radioimmunoconjugated with $^{177}Lu$. The in vitro high immunoactivity of this radioimmunoconjugate was determined by a cell binding assay. In addition, the antibody's tumor targeting ability was demonstrated with very high uptake in tumors. This radioimmunoconjugate is applicable to therapy in human colon cancer with highly expressed CD44.

Ruminal microbial responses in fermentation characteristics and dry matter degradability to TDN level of total mixed ration

  • Lee, Seung-Uk;Jo, Jin-Ho;Park, Sung-Kwon;Choi, Chang-Weon;Jeong, Jun;Chung, Ki-Young;Chang, Sun-Sik;Li, Xiang Zi;Choi, Seong-Ho
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.80-86
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    • 2016
  • An in vitro trial was conducted to examine the effects of total mixed rations (TMR) on fermentation characteristics and effective degradability (ED) by rumen microbes. Three TMR diets were growing period TMR (GR-TMR, 67% TDN), early fattening period TMR (EF-TMR, 75.4% TDN) and late fattening TMR (LF-TMR, 80% TDN). Three TMR diets (3 g of TMRs in each incubation bottles) was added to the mixed culture solution of stained rumen fluid with artificial saliva (1:1, v/v) and incubated anaerobically for 48 hours at $39^{\circ}C$. The pH in all incubation solutions tended to decrease up to 48h, but the opposite results were found in concentration of total gas production, ammonia-N and total VFA in all incubations.The total gas production (p<0.05) in LF-TMR was highest compared with those of other diets. Also, concentration of total VFA was tended to increase in LF-TMR compared with other TMR diets in all incubations. The EDDM in both EF-TMR and LF-TMR was tended to high compared with GR-TMR (p=0.100). In this in vitro trials, concentration of propionate in all incubation solution was not affected by increased concentration of TDN. The results of the present in vitro study indicate that TMR may provide more favorable condition for nutrient digestion both in the rumen.

Effects of Chromium Picolinate (CrP) on Growth Performance, Carcass Characteristics and Serum Traits in Growing-Finishing Pigs

  • Min, J.K.;Kim, W.Y.;Chae, B.J.;Chung, I.B.;Shin, I.S.;Choi, Y.J.;Han, I.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.1
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    • pp.8-14
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    • 1997
  • An experiment was carried out to evaluate the effects of feeding graded levels of chromium in the form of chromium picolinate on growth performance, blood components, carcass grade, in vitro lipogenesis and lipolysis, and in vitro protein degradation and synthesis in growing-finishing pigs. There were no significant differences for daily weight gain, feed intake and feed conversion among treatments during growing phase, while in the finishing phase, feed intake was lower in groups fed diets with 200 ppb chromium than in other treatment (p < 0.1). Feed conversion was improved in the groups fed diets with chromium compared with control. Carcass weight was similar among treatments while carcass length was longer in groups fed diets with 200 ppb chromium (p < 0.05). Thinner carcass fat was found with groups fed diets with chromium compared to control. Three A grade of carcasses were from groups fed chromium compared to control. No significant differences were observed with blood glucose, insulin, total cholesterol, triglycerides and non-esterified fatty acid at 60kg body weight. While, at 100 kg body weight, blood triglyceride was lower in groups with 200 and 400 ppb chromium but higher in groups with 100 ppb chromium (p < 0.05). In vitro lipolysis and protein synthesis in adipose tissues were increased as dietary chromium was increased from 0 to 200 ppb (p < 0.1). As a result, 200 ppb chromium in a growing-finishing diet could improve feed efficiency and carcass traits; an increase to 400 ppb has no further effect.

Cytolytic Activities of Taxol on Neural Stem Cells

  • Lee, In-Soo;Han, Hye-Eun;Lee, Hye-Young;Kim, Seung-U.;Kim, Tae-Ue
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.273-278
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    • 2007
  • Stem cells have been the subject of increasing scientific interest because of their utility in numerous biomedical applications. Stem cells are capable of renewing themselves; that is, they can be continuously cultured in an undifferentiated state, giving rise to more specialized cells of the human body. Therefore, stem cells are an important new tools for developing unique, in vitro model systems to test drugs and chemicals and a potential to predict or anticipate toxicity in humans. In the present study, in vitro cultured F3 immortalized human neural stem cell line and in vivo adult Sprague Dawley rats was used to evaluate the cytotoxicity of anticancer drug paclitaxel. In vitro apoptotic activity of paclitaxel was evaluated in F3 cell line by a MTT assay and DAPI test. The cell death was induced with the treatment of 20 nM paclitaxel and chromatin degradation was detected by DAPI staining, which was analyzed by fluorescent microscope. In vivo studies, we also observed nestin immunoreactivity on subventricular zone, which is stem cell rich region in the adult brain of the SD rat. Immunofluorescent staining result shows that pixel intensities of nestin were decreased in a dose dependent manner. These results suggest that paclitaxel is able to induce cytotoxic activity both in F3 neural stem cell line and neural stem cell in SD rat brain.

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Forward Gene Mutation Assay of Seven Benzophenone-type UV Filters using L5178Y Mouse Lymphoma Cell

  • Jeon, Hee-Kyung;Sarma, Sailendra Nath;Kim, Youn-Jung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.3 no.1
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    • pp.23-30
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    • 2007
  • The effects of high energy short wave solar radiation on human skin have received much publicity as the major cause of accelerated skin ageing and of skin cancers. To meet public demand, the cosmetic industry has developed sun protection factor products, which contain a variety of so-called "UV filters", among others benzophenone (BP) and its metabolites are the widely used UV filters. UV filters are also used to prevent UV light from damaging scents and colors in a variety of cosmetics products and to protect of plastic products against light-induced degradation. There are many variants of BP in use. In this respect, to regulate and to evaluate the hazardous effect of BP-type UV filters will be important to environment and human health. The genotoxicity of 7 BP-type UV filters was evaluated in L5178Y $(tk^{+/-})$ mouse lymphoma cells in vitro. BP, benzhydrol, 4-hydroxybenzophenone 2-hydroxy-4-methoxybenzophenone and 2, 4-dihydroxybenzophenone did not induce significant mutation frequencies both in the presence and absence of metabolic activation system. 2, 2'-Dihydroxy-4-methoxybenzophenone appeared the positive results at the highest dose, i.e. 120.4 ${\mu}g/mL$ only in the absence of metabolic activation system. And also, 2, 3, 4-trihydroxybenzophenone revealed a significant increase of mutation frequencies in the range of 138.1-207.2 ${\mu}g/mL$ in the absence of metabolic activation system and 118.3-354.8 ${\mu}g/mL$ in the presence of metabolic activation system. Through the results of MLA with 7 BP-type UV filters in L5178Y cells in vitro, we may provide the important clues on the genotoxic potentials of these BP-type UV filters.

Wheat phytase can alleviate the cellular toxic and inflammatory effects of lipopolysaccharide

  • An, Jeongmin;Cho, Jaiesoon
    • Journal of Animal Science and Technology
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    • v.63 no.1
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    • pp.114-124
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    • 2021
  • The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on in vitro toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. In vitro toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h (p < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the in vitro toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively (p < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% (p < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.

Productivity and Nutritive Values of Different Fractions of Oil Palm (Elaeis guineensis) Frond

  • Islam, M.;Dahlan, I.;Rajion, M.A.;Jelan, Z.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.8
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    • pp.1113-1120
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    • 2000
  • Productivity, nutrient contents, in vitro gas production and in sacco degradability of different fractions and whole OPF were determined to assess the feeding value of OPF as a ruminant feed. An in vivo digestibility trial was also carried out using goat. Freshly harvested OPF was randomly collected, partitioned and weighed. An OPF from 21 years older palm weighed 13.4 kg and the annual fresh matter yield of petiole, leaflet and midrib was 46.5, 11.8 and 3.4 ton/ha, respectively. Leaflet contained 439, 926, 698, 501, 168, 196, 748 and 52 (g/kg) of dry matter (DM), organic matter (OM), neutral detergent fiber (NDF), acid detergent fiber (ADF), cellulose (CE), hemicellulose (HC), total carbohydrate (TC) and non fiber carbohydrate (NFC), respectively. Petiole contained lower (p<0.01) DM, CP and EE contents than leaflet. Organic matter, CE and TC contents were higher (p<0.01) in petiole compared to leaflet. Silica and lignin contents were highest (p<0.01) in midrib followed by leaflet, whole OPF and least in petiole. The Ca, P, Na, K and Mg contents (g/100 g DM) of leaflet were 0.529, 0.182, 0.039, 0.876, and 0.168, respectively. In vitro DM digestibility (g/100 g) at 48 h of leaflet, petiole and midrib was 32.7, 38.7 and 30.2, respectively. The in sacco DM degradation (g/100 g) at 48 h of leaflet was higher than that of whole OPF, petiole and midrib. The in vivo digestibility of DM, OM, CP and ADF of whole OPF was 52, 56, 43 and 26%, respectively. It can be concluded that leaflet is the most nutritious fraction of OPF and midrib is the least. The nutrient content and digestibility of the whole OPF showed that OPF could be an alternative roughage source for ruminant diets.

Effect of Hijikia fusiforme extracts on degenerative osteoarthritis in vitro and in vivo models

  • Kwon, Han Ol;Lee, Minhee;Kim, Ok-Kyung;Ha, Yejin;Jun, Woojin;Lee, Jeongmin
    • Nutrition Research and Practice
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    • v.10 no.3
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    • pp.265-273
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    • 2016
  • BACKGROUND/OBJECTIVES: The inhibitory effect of Hijikia fusiforme (HF) extracts on degenerative osteoarthritis was examined in primary cultured rat cartilage cells and a monosodium iodoacetate (MIA)-induced osteoarthritis rat model. MATERIALS/METHODS: In vitro, cell survival and the expression of matrix metalloproteinases (MMPs), collagen type I, collagen type II, aggrecan, and tissue inhibitor of metalloproteinases (TIMPs) was measured after $H_2O_2$ ($800{\mu}M$, 2 hr) treatment in primary chondrocytes. In vivo animal study, osteoarthritis was induced by intra-articular injection of MIA into knee joints of rats, and then RH500, HFE250 and HFE500 were administered orally once a day for 28 days. To determine the anti-inflammatory effects of HFE, nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) expression were measured. In addition, real-time PCR was performed to measure the genetic expression of MMPs, collagen type I, collagen type II, aggrecan, and TIMPs. RESULTS: In the in vitro assay, cell survival after $H_2O_2$ treatment was increased by HFE extract (20% EtOH). In addition, anabolic factors (genetic expression of collagen type I, II, and aggrecan) were increased by HFE extract (20% EtOH). However, the genetic expression of MMP-3 and 7, known as catabolic factors were significantly inhibited by treatment with HFE extract (20% EtOH). In the in vivo assay, anabolic factors (genetic expression of collagen type I, II, aggrecan, and TIMPs) were increased by oral administration of HFE extract. However, the genetic expression of MMP-3 and 7, known as catabolic factors, and production of NO and $PGE_2$ were significantly inhibited by treatment with oral administration of HFE extract. CONCLUSION: HFE extract inhibited articular cartilage degeneration through preventing extracellular matrix degradation and chondrocyte injury.