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Pseudomonas oleovorans의 유가식 배양에 의한 medium chain length Polyhydroxyalkanoates (MCL-PHA) 생산

  • Kim, Beom-Su;Im, Hui-Yeon
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.207-210
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    • 2000
  • Pseudomonas oleovorans was cultivated to produce medium chain length polyhydroxyalkanoates (MCL-PHA) fram octanoic acid and ammonium nitrate as carbon and nitrogen source, respectively, by a pH-stat fed-batch culture technique. The octanoate concentration of the culture broth was maintained below 4 g/L by feeding the mixture of octanoic acid and ammonium nitrate when the culture pH rose above high limit. The effect of the ratio of octanoic acid to ammonium nitrate (C/N ratio) in the feed on the PHA production was examined. The final cell concentrations of 62.5, 54.7, and 9.5 g/L, PHA contents of 62.9, 75.1, and 67.6% of dry cell weight, and productivities of 1.03, 0.632, and 0.161 g/L/h were obtained when the C/N ratio in the feed were 10, 20, and 100 g octanoic acid/g ammonium nitrate, respectively.

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Root Bark extract of Morus alba L. Suppressed the Migration and Invasion of HCT116 Human Colorectal Carcinoma Cells (HCT116 인체 대장암 세포주에서 상백피 추출물에 의한 전이 억제 효과)

  • Park, Shin-Hyung;Park, Hyun-Ji
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.35 no.5
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    • pp.177-184
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    • 2021
  • The root bark of Morus alba L. (MA) used in traditional oriental medicine for the treatment of pulmonary diseases exerts various pharmacological activities including anticancer effects. In the current study, we investigated the effects of MA on the migration and invasion of colorectal carcinoma cells. Results from a transwell assay showed that the methylene chloride extract of MA (MEMA) suppressed the migration and invasion of HCT116 human colorectal carcinoma cells in a concentration-dependent manner. MEMA reduced both mRNA and protein levels of matrix metalloproteinase (MMP)-9, but did not suppress the expression of MMP-2 in HCT116 cells. As a molecular mechanism, MEMA inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including ERK, JNK and p38, in a dose-dependent manner. In addition, MEMA dephosphorylated both Src and signal transducer and activator of transcription 3 (STAT3) in HCT116 cells. Taken together, we demonstrate that MEMA suppressed the migration and invasion capacity of HCT116 human colorectal cancer cells by downregulation of MMP-9 and inactivation of both MAPKs and Src/STAT3 signaling pathway.

Based on morphology and molecular data, Palisada rigida comb. nov. and Laurencia decussata comb. et stat. nov. (Rhodophyta, Rhodomelaceae) are proposed

  • Metti, Yola
    • ALGAE
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    • v.37 no.1
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    • pp.15-32
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    • 2022
  • Inspecting herbaria collections of Laurencia rigida highlighted frequent misidentifications between L. rigida and L. heteroclada f. decussata, two poorly studied taxa from Australia. Recent collections of DNA material, including from topotype material, allowed for re-examination of these two taxa using molecular techniques. Detailed morphological and molecular analyses based on two markers (rbcL and COI-5P) strongly supported these two taxa as being distinct from each other and requiring nomenclatural changes. Comprehensive morphological analyses highlighted features useful for accurate identifications. Interestingly, L. rigida was found to belong to the genus Palisada with evidence from both the morphology and molecular data. Therefore, this study proposed recognizing L. rigida as Palisada rigida comb. nov. Molecular data for L. heteroclada f. decussata on the other hand supported its separation from L. heteroclada, with too great a molecular distance to be considered a variety. Morphological characters that best separated P. rigida from L. decussata included seven characters; number of pericentral cells per vegetative axial segment, the presence of secondary pit connections, the presence of lenticular thickenings, tetrasporangia alignment, the presence of corps en cerise, holdfast morphology, and overall plant shape. Morphologically, L. heteroclada f. decussata was also separated from L. heteroclada, particularly by the following characteristics; ultimate branchlets morphologies, lower order branch lengths, primary axis and holdfast morphologies. Therefore, it was proposed that L. heteroclada f. decussata is recognized at a species level as L. decussata comb. et stat. nov.

Nuclear Receptor PPARα Agonist Wy-14,643 Ameliorates Hepatic Cell Death in Hepatic IKKβ-Deficient Mice

  • Kim, Taehyeong;Wahyudi, Lilik Duwi;Gonzalez, Frank J.;Kim, Jung-Hwan
    • Biomolecules & Therapeutics
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    • v.25 no.5
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    • pp.504-510
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    • 2017
  • Inhibitor of nuclear factor kappa-B kinase beta ($IKK{\beta}$) plays a critical role in cell proliferation and inflammation in various cells by activating $NF-{\kappa}B$ signaling. However, the interrelationship between peroxisome proliferator-activated receptor ${\alpha}$ ($PPAR{\alpha}$) and $IKK{\beta}$ in cell proliferation is not clear. In this study, we investigated the possible role of $PPAR{\alpha}$ in the hepatic cell death in the absence of $IKK{\beta}$ gene using liver-specific Ikkb-null ($Ikkb^{F/F-AlbCre}$) mice. To examine the function of $PPAR{\alpha}$ activation in hepatic cell death, wild-type ($Ikkb^{F/F}$) and $Ikkb^{F/F-AlbCre}$ mice were treated with $PPAR{\alpha}$ agonist Wy-14,643 (0.1% w/w chow diet) for two weeks. As a result of Wy-14,643 treatment, apoptotic markers including caspase-3 cleavage, poly (ADP-ribose) polymerase (PARP) cleavage and TUNEL-positive staining were significantly decreased in the $Ikkb^{F/F-AlbCre}$ mice. Surprisingly, Wy-14,643 increased the phosphorylation of p65 and STAT3 in both Ikkb and $Ikkb^{F/F-AlbCre}$ mice. Furthermore, BrdU-positive cells were significantly increased in both groups after treatment with Wy-14,643. Our results suggested that $IKK{\beta}-derived$ hepatic apoptosis could be altered by $PPAR{\alpha}$ activation in conjunction with activation of $NF-{\kappa}B$ and STAT3 signaling.

Anti-Inflammatory Effects of Prunus mume Mixture in Colitis Induced by Dextran Sodium Sulfate (매실 혼합물이 DSS로 유도된 염증성 장질환 동물모델의 면역조절에 미치는 활성)

  • Jin, Hai-Lan;Lee, Bo-Ram;Lim, Kyung-Jik;Debnath, Trishna;Shin, Heung-Mook;Lim, Beong-Ou
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.1
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    • pp.16-23
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    • 2011
  • This study was conducted to investigate the anti-inflammatory effects of Pruns mume, Schisandra chinensis, Chaenomeles sinensis-- Prunus mume mixtrue (PM) treatment on colitis induced in mice by dextran sodium sulfate (DSS) treatment. A total of 25 male BALB/c mice (average weight $20.7\;{\pm}\; 1.6 \;g$) were divided into 5 treatment groups and fed a commercial diet (A), PM administration (B), commercial diet + induced colitis by DSS (C), PM administration + induced colitis by DSS (D) and sulfasalazine + induced colitis by DSS (E). We found that PM treatment (D) and sulfasalazine (E) decreased the expression of $TNF-{\alpha}$ and COX-2 compared to the DSS-induced colitis group (C). The expression of IL-4, STAT6, $IFN-{\gamma}$, STAT1 was decreased in group D and group E compared to the colitis group (C), COX-2 and STAT1 were more decreased in group D. The serum IgE levels decreased in the PM treatment groups (C and D) compared to the non-PM treatment groups (A and B) although there was no significant difference between the PM treatment groups. It is notable that a therapeutic application of the PM extracts ameliorated DSS-induced colitis in mice.

Effect of all-trans retinoic acid on casein and fatty acid synthesis in MAC-T cells

  • Liao, Xian-Dong;Zhou, Chang-Hai;Zhang, Jing;Shen, Jing-Lin;Wang, Ya-Jing;Jin, Yong-Cheng;Li, Sheng-Li
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.1012-1022
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    • 2020
  • Objective: Caseins and fatty acids of milk are synthesized and secreted by the epithelial cells of the mammary gland. All-trans retinoic acid (ATRA), an active metabolite of vitamin A, has been shown to promote mammary development. This study was conducted to determine the effect of ATRA on casein synthesis and fatty acid composition in MAC-T cells. Methods: MAC-T cells were allowed to differentiate for 4 d, treated with ATRA (0, 1.0, 1.5, and 2.0 μM), and incubated for 3 d. We analyzed the fatty acid composition, the mRNA expression of casein and fatty acid synthesis-related genes, and the phosphorylation of casein synthesis-related proteins of MAC-T cells by gas chromatography, quantitative polymerase chain reaction, and western blotting, respectively. Results: In MAC-T cells, ATRA increased the mRNA levels of αS1-casein and β-casein, janus kinase 2 (JAK2) and E74-like factor 5 of the signal transducer and activator of transcription 5 β (STAT5-β) pathway, ribosomal protein S6 kinase beta-1 (S6K1) and eukaryotic translation initiation factor 4E binding protein 1 of the mammalian target of rapamycin (mTOR) pathway, inhibited the mRNA expression of phosphoinositide 3-kinase and eukaryotic initiation factor 4E of the mTOR pathway, and promoted the phosphorylation of STAT5-β and S6K1 proteins. Additionally, ATRA increased the de novo synthesis of fatty acids, reduced the content of long-chain fatty acids, the ratio of monounsaturated fatty acids to saturated fatty acids (SFA), the ratio of polyunsaturated fatty acids (PUFA) to SFA, and the ratio of ω-6 to ω-3 PUFA. The mRNA levels of acetyl-CoA carboxylase 1, fatty acid synthase, lipoprotein lipase, stearoyl-CoA desaturase, peroxisome proliferator-activated receptor gamma, and sterol regulatory element-binding protein 1 (SREBP1) were enhanced by ATRA. Conclusion: ATRA promotes the synthesis of casein by regulating JAK2/STAT5 pathway and downstream mTOR signaling pathway, and it improves the fatty acid composition of MAC-T cells by regulating SREBP1-related genes.

Production of ${\beta}-Galactosidase$ with High Transgalactosylation Activity by Bacillus sp. A4442 Mutant (변이주 Bacillus sp. A4442에 의한 갈락토스 전이활성이 높은 ${\beta}-Galactosidase$의 생산)

  • Choi, Kyung-Ho;Yang, Sung-Joon;Kim, Min-Hong;Han, Keum-Soo;Yang, Ji-Won;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.507-511
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    • 1995
  • In an attempt to improve the productivity of ${\beta}-galactosidase$ from Bacillus sp. A1, which was isolated from soil and has remarkably higher transgalactosylation activity than lactose hydrolysis activity, a chemical mutation procedure using N-methyl-N'-nitro-N-nitrosoguanidine followed by selection was conducted. The final selection, designated as Bacillus sp. A4442, turned out to show a substantially increased enzyme productivity. Catabolite repression by glucose and lactose requirement as an inducer for the enzyme biosynthesis, which were shown in the parent strain, was markedly diminished; instead it was found out that galactose acts as another inducer. Because pH of medium, one of the most important factors for cell growth as well as enzyme production, is closely related with the sugar concentration during culture, it was kept in the optimum range of $6.5{\sim}7.5$; for this the initial glucose concentration was adjusted to be 0.5% which was thereafter maintained by the controlled pumping-in of lactose using the pH-stat technique. By doing so, we were able to increase the productivity of ${\beta}-galactosidase$ with high transgalactosylation activity up to $44\;unit/m{\ell}-broth$.

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MiR-30a-5p and miR-153-3p regulate LPS-induced neuroinflammatory response and neuronal apoptosis by targeting NeuroD1

  • Choi, Hye-Rim;Ha, Ji Sun;Kim, Eun-A;Cho, Sung-Woo;Yang, Seung-Ju
    • BMB Reports
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    • v.55 no.9
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    • pp.447-452
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    • 2022
  • Neurogenic differentiation 1 (NeuroD1) is an essential transcription factor for neuronal differentiation, maturation, and survival, and is associated with inflammation in lipopolysaccharide (LPS)-induced glial cells; however, the concrete mechanisms are still ambiguous. Therefore, we investigated whether NeuroD1-targeting miRNAs affect inflammation and neuronal apoptosis, as well as the underlying mechanism. First, we confirmed that miR-30a-5p and miR-153-3p, which target NeuroD1, reduced NeuroD1 expression in microglia and astrocytes. In LPS-induced microglia, miR-30a-5p and miR-153-3p suppressed pro-inflammatory cytokines, reactive oxygen species, the phosphorylation of c-Jun N-terminal kinase, extracellular-signal-regulated kinase (ERK), and p38, and the expression of cyclooxygenase and inducible nitric oxide synthase (iNOS) via the NF-κB pathway. Moreover, miR-30a-5p and miR-153-3p inhibited the expression of NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasomes, NLRP3, cleaved caspase-1, and IL-1β, which are involved in the innate immune response. In LPS-induced astrocytes, miR-30a-5p and miR-153-3p reduced ERK phosphorylation and iNOS expression via the STAT-3 pathway. Notably, miR-30a-5p exerted greater anti-inflammatory effects than miR-153-3p. Together, these results indicate that miR-30a-5p and miR-153-3p inhibit MAPK/NF-κB pathway in microglia as well as ERK/STAT-3 pathway in astrocytes to reduce LPS-induced neuronal apoptosis. This study highlights the importance of NeuroD1 in microglia and astrocytes neuroinflammation and suggests that it can be regulated by miR-30a-5p and miR-153-3p.

Activation of Cabbage Phospholipase D by Polyamines (폴리아민에 의한 양배추 포스포리파제 D의 활성화)

  • Eun-Hie Koh
    • Journal of the Korean Chemical Society
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    • v.47 no.5
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    • pp.466-471
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    • 2003
  • The effect of polyamines on the cabbage phospholipase D(PLD) activity was investigated. The PLD activity was determined by pH-stat titration of phosphatidic acid, one of the enzymatic reaction product, using phosphatidyl choline small unilamellar vesicles as a substrate. The cabbage PLD was activated approximately 4 fold by spermine at 1 mM concentration. This spermine effect appears to be similar to the previous report on the PLD activation of rat brain mitochondrial fraction. It was also found that cationic polypetides such as polylysine and polyhistidine exerted a marked enhancement effect on the cabbage PLD. Particularly polyhistidine exerted approximately 5.5 fold enhancement effect at 0.062 mM concentration. The polyamine effect on the cabbage PLD was reexamined in the phosphatidylcholine/sodium dodecyl sulfate mixed micellar system. The relevance of polyamine effect on PLD activity is discussed in relation to the active site of PLD.

Immunoregulatory Effects of Saengshik on DSS-Induced Inflammatory Bowel Disease in Mouse Model System (DSS로 유도된 염증성 장 질환 마우스 동물모델에서 생식이 장관 임파조직내 면역조절 기능에 미치는 영향)

  • Lim, Beong-Ou;Jeong, Yong-Jun;Park, Mi-Hyoun;Kim, Jong-Dai;Hwang, Sung-Joo;Yu, Byung-Pal
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.1
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    • pp.32-42
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    • 2007
  • This study was conducted on the immunoregulatory effect of Saengshik on gut-associated lymphoid tissue with inflammatory bowel disease. Although the contents of IgA increased in mesenteric lymph node, IgE content was suppressed by Saengshik. The same results were found in spleen, but IgA and IgE responses were very weak. Concentration of fecal IgA was high from the first day through the third day in Saengshik group. In DSS + Saengshik group, concentration of IgA was high till the 2nd day and it maintained the highest level among the test groups on 5th day. Concentration of IFN-gamma and IL-2 was the highest in the Saengshik group, but the concentration of TNF-alpha was lower in DSS + Saengshik compared to DSS. The expressions of STAT1 in Saengshik group were high, while those of STAT6 were low According to these findings, Saengshik exhibited effectiveness via increasing the IgA production, suppressing the IgE production, followed by inhibiting the production of IL-4 and IL-10. Saengshik also strengthened the immune system and alleviated injury in DSS -induced inflammation.