• Title/Summary/Keyword: RNA processing

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Inhibition of Pancreatic Lipase Activity and Adipocyte Differentiation in 3T3-L1 Cells Treated with Purple Corn Husk and Cob Extracts (자색옥수수 포엽과 속대 추출물의 리파아제 저해활성 및 3T3-L1 지방전구세포에서의 지방분화 억제효과)

  • Lee, Ki Yeon;Hong, Soo Young;Kim, Tae Hee;Kim, Jai Eun;Park, A-Reum;Noh, Hee Sun;Kim, Si Chang;Park, Jong Yeol;Ahn, Mun Seob;Jeong, Won Jin;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.131-139
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    • 2018
  • Our review begins with the maize hybrid for grain, called 'Seakso 1,' which was developed in 2008 by the Gangwon Agricultural Research and Extension Services in Korea, and subsequently registered in 2011. In this study, we aimed to investigate the lipid metabolic enzyme activity and inhibitory effect on the adipocyte differentiation, in 3T3-L1 cells of the identified Seakso 1 corn husk and cob extracts (EHCS). We investigated the pancreatic lipase inhibitory effect and anti-adipogenic effect of EHCS.The lipid accumulation and adipocyte differentiation were measured by the procedure of Oil Red O staining, Real-time PCR and the Western blot analysis. The pancreatic lipase inhibitory activity of EHCS was measured at higher levels than those of the positive control (orlistat) at 100, 500, and $1,000{\mu}g/mL$. In particular, EHCS was noted as being significantly inhibited and including a measured adipocyte differentiation and lipid accumulation, when treated during the adipocyte differentiation process in 3T3-L1 cells. Based on the Oil Red O staining, EHCS inhibited lipid accumulation at 19.19%, 33.30% at $1000{\mu}g/mL$, $2000{\mu}g/mL$, respectively. The real-time PCR and Western blot analysis showed that EHCS significantly decreased in the mRNA expression and protein level of obesity-related factors, such as peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). This study potentially suggests that the Saekso 1 corn husk and cob extracts may improve lipid metabolism and reduce lipid accumulation.

Identification and Functional Analysis of Escherichia coli RNase E Mutants (Escherichia coli 리보핵산 내부분해효소 RNase E의 돌연변이체 선별 및 특성분석)

  • Shin, Eun-Kyoung;Go, Ha-Young;Kim, Young-Min;Ju, Se-Jin;Lee, Kang-Seok
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.325-330
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    • 2007
  • RNase E is an essential Escherichia coli endoribonuclease that plays a major role in the decay and processing of a large fraction of RNAs in the cell and expression of N-terminal domain consisted of 1-498 amino acids (N-Rne) is sufficient to support normal cellular growth. By utilizing these properties of RNase E, we developed a genetic system to screen for amino acid substitutions in the catalytic domain of the protein (N-Rne) that lead to various phenotypes. Using this system, we identified three kinds of mutants. A mutant N-Rne containing amino acid substitution in the S1 domain (I6T) of the protein was not able to support survival of E. coli cells, and another mutant N-Rne with amino acid substitution at the position 488 (R488C) in the small domain enabled N-Rne to have an elevated ribonucleolytic activity, while amino acid substitution in the DNase I domain (N305D) only enabled N-Rne to support survival of E. roli cells when the mutant N-Rne was over-expressed. Analysis of copy number of ColEl-type plasmid revealed that effects of amino acid substitution on the ability of N-Rne to support cellular growth stemmed from their differential effects on the ribonucleolytic activity of N-Rne in the cell. These results imply that the genetic system developed in this study can be used to isolate mutant RNase E with various phenotypes, which would help to unveil a functional role of each subdomain of the protein in the regulation of RNA stability in E. coli.

Effects of Jeju Citrus unshiu Peel Extracts Before and After Bioconversion with Cytolase on Anti-Inflammatory Activity in RAW264.7 Cells (면역세포에서 Bioconversion 전후 제주 감귤 과피 추출물의 항염증 효과)

  • Seo, Jieun;Lim, Heejin;Chang, Yun-Hee;Park, Hye-Ryeon;Han, Bok-Kyung;Jeong, Jung-Ky;Choi, Kyoung-Sook;Park, Su-Beom;Choi, Hyuk-Joon;Hwang, Jinah
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.3
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    • pp.331-337
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    • 2015
  • Citrus and its peels, which are by-products from juice and/or jam processing, have long been used in Asian folk medicine. Citrus peels show an abundant variety of flavanones, and these flavanones have glycone and aglycone forms. Aglycones are more potent than glycones with a variety of physiological functions since aglycone absorption is more efficient than glycones. Bioconversion with cytolase converted narirutin and naringin into naringenin and hesperidin into hesperetin. Therefore, this study aimed to investigate the anti-oxidant and anti-inflammatory effects of bioconversion of Citrus unshiu (CU) peel extracts with cytolase (CU-C) in RAW264.7 cells. HPLC chromatograms showed that CU and CU-C had 23.42% and 29.39% total flavonoids, respectively. There was substantial bioconversion of narirutin to naringenin and of hesperidin to hesperetin. All citrus peel extracts showed DPPH scavenging activities in a dose-dependent manner, and CU-C was more potent than intact CU. RAW264.7 cells were pre-treated with $0{\sim}500{\mu}g/mL$ of citrus peel extracts for 4 h and then stimulated by $1{\mu}g/mL$ of lipopolysaccharide (LPS) for 8 h. All citrus peel extracts showed decreased mRNA levels and protein expression of LPS-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in a dose-dependent manner. Especially, CU-C markedly inhibited mRNA and protein expression of iNOS and COX-2 compared to intact citrus peel extracts. All citrus peel extracts showed decreased NO production by iNOS activity. This result suggests that bioconversion of citrus peel extracts with cytolase may provide potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by boosting the anti-inflammatory effects of citrus peels.

Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Dna2 Helicase/endonuclease Interacts with a Novel Protein YHR122W Protein in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 Dna2 helicase/endonuclease와 YHR122W 단백질의 상호작용)

  • Lee, Hyun-Sun;Choi, Do-Hee;Kwon, Sung-Hoon;Kim, Na-Yeon;Lee, In-Hwan;Kim, Hyun-Jung;Bae, Sung-Ho
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.1-6
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    • 2006
  • Saccharomyces cerevisiae Dna2 helicase/endonuclease plays an essential role in removing DNA primers during Okazaki fragment processing in eukaryotic DNA replication. Genome-wide scale co-immunoprecipitation experiments predicted that Dna2 interacts with a novel protein YHR122W (1). In this study, we observed that overexpression of YHR122W gene suppressed the temperature-sensitive phenotype of $dna2\Delta405N$ mutation. To investigate direct interaction between these two proteins, a histidine-tagged recombinant YHR122W protein was expressed and purified from E. coli. Physical interaction between the purified YHR122W and Dna2 proteins was detected by enzyme-linked immunosorbent assays. Further more, the complex formation was most efficient at physiological salt concentration, 150 mM NaCl. The genetic and physical interactions between YHR122W and Dna2 shown in this study suggest that the biological functions of these two proteins may be closely related each other.

Ethyl Acetate Extract of Korean Rice Wine Lees Inhibits IgE-Mediated Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells and Passive Cutaneous Anaphylaxis in Mice (주박 에틸아세테이트 추출물의 항알러지 효과)

  • Kang, Yeo-Jin;Park, Sae-Jin;Bae, Ki-Ho;Yoo, Jung-Min;Pyo, Hyeong-Bae;Choi, Ji-Ho;Kim, Tack-Joong
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1364-1369
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    • 2011
  • Mast cells, the central effector cells involved in the allergic response, release histamine, arachidonic acid, and proinflammatory cytokines. We investigated the effect of the ethyl acetate fraction (EA), derived from Korean rice wine lees, on RBL-2H3 cell degranulation and passive cutaneous anaphylaxis in an animal model. The EA fraction suppressed the release of beta-hexosaminidase, a marker of degranulation, and the mRNA expression of interleukin-3 (IL-3) and IL-13. EA also successfully suppressed the passive cutaneous anaphylaxis (PCA) reaction in mice in a dose-dependent manner. These results suggest that EA can inhibit mast cell degranulation through the inhibition of IL-3 and IL-13 mRNA expression, and that EA may potentially serve as an anti-allergic agent.

Isolation and Characterization of Lactic Acid Bacteria with Angiotensin-Converting Enzyme Inhibitory and Antioxidative Activities (안지오텐신 전환효소 저해 활성 및 항산화 활성을 가진 젖산균의 분리 및 특성)

  • Park, Sung-Bo;Kim, Jeong-Do;Lee, Na-Ri;Jeong, Jin-Ha;Jeong, Seong-Yun;Lee, Hee-Seob;Hwang, Dae-Youn;Lee, Jong-Sup;Son, Hong-Joo
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1428-1433
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    • 2011
  • In this study, we isolated and characterized plant-associated lactic acid bacteria which are able to produce angiotensin-converting enzyme (ACE) inhibitory and antioxidative activities. Five lactic acid bacteria were isolated from plants (grape and leek), a plant-associated fermentative product (Kimchi) and Korean traditional alcohol (Dongdongju). Strains K-1 and K-21 from Kimchi, strain L-5 from leek, strain G-3 from grape, and strain D-3 from Dongdongju were identified as Pediococcus pentosaceus, Lactobacillus plantarum, Weissella cibaria, L. plantarum, and L. brevis, respectively, by 16S rRNA gene analysis. ACE inhibitory activities of isolated strains ranged from 44.3 to 71.9% in the MRS broth. G-3, L-5 and K-1 strains especially showed high ACE inhibitory activities (59.8-98.69%) in the MRS broth containing skim milk. DPPH radical scavenging activities of the strains were in the range of 42.5-82.7%. All strains showed varying levels of resistance in artificial gastric fluid (pH 2.5), retaining viability ranging from 42.2 to 88.1% after 3 hr of incubation. All strains showed high resistance to 0.3% oxgall after 24 hr of incubation; survival rates were in the range of 55.4-112.8%. Isolated strains were found to be antagonistic to some pathogens including Pseudomonas aeruginosa.

Resveratrol Inhibits $GABA_C$ ${\rho}$ Receptor-Mediated Ion Currents Expressed in Xenopus Oocytes

  • Lee, Byung-Hwan;Choi, Sun-Hye;Hwang, Sung-Hee;Kim, Hyeon-Joong;Lee, Joon-Hee;Nah, Seung-Yeol
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.2
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    • pp.175-180
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    • 2013
  • Resveratrol is a phytoalexin found in grapes, red wine, and berries. Resveratrol has been known to have many beneficial health effects, such as anti-cancer, neuroprotective, anti-inflammatory, and life-prolonging effects. However, relatively little is known about the effects of resveratrol on the regulation of ligand-gated ion channels. We have previously reported that resveratrol regulates subsets of homomeric ligand-gated ion channels such as those of 5-$HT_{3A}$ receptors. The ${\gamma}$-aminobutyric $acid_C$($GABA_C$) receptor is mainly expressed in retinal bipolar cells and plays an important role in visual processing. In the present study, we examined the effects of resveratrol on the channel activity of homomeric $GABA_C$ receptor expressed in Xenopus oocytes injected with cRNA encoding human $GABA_C$ ${\rho}$ subunits. Our data show that the application of GABA elicits an inward peak current ($I_{GABA}$) in oocytes that express the $GABA_C$ receptor. Resveratrol treatment had no effect on oocytes injected with $H_2O$ or with $GABA_C$ receptor cRNA. Co-treatment with resveratrol and GABA inhibited $I_{GABA}$ in oocytes with $GABA_C$ receptors. The inhibition of $I_{GABA}$ by resveratrol was in a reversible and concentration-dependent manner. The $IC_{50}$ of resveratrol was $28.9{\pm}2.8{\mu}M$ in oocytes expressing $GABA_C$ receptor. The inhibition of $I_{GABA}$ by resveratrol was in voltage-independent and non-competitive manner. These results indicate that resveratrol might regulate $GABA_C$ receptor expression and that this regulation might be one of the pharmacological actions of resveratrol on the nervous system.

Nitric Oxide on the MMP-2 expression by human gingival fibroblasts (치은섬유아세포의 MMP 발현에 대한 Nitric Oxide의 영향)

  • Shin, In-Sik;Yoon, Sang-Oh;Chung, Hyun-Ju;Koh, Jung-Tae
    • Journal of Periodontal and Implant Science
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    • v.33 no.2
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    • pp.277-288
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    • 2003
  • It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HTl080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP , suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

Differences in swine gut microbiota in southern region of Republic of Korea (한국 남부 지역별 돼지 장내 미생물생태 비교분석)

  • Kim, Jungman;Guevarra, Robin B.;Nguyen, Son G.;Unno, Tatsuya
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.81-85
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    • 2015
  • Since the banning of antibiotic growth promoters (AGPs), the death of livestock has been increased, thus there is a strong demand for AGP-alternatives. Modulation of gut microbiota has been reported to affect host physiological functions and suggested to be a novel approach for developing AGP-alternatives. However, little has been understood about livestock gut microbiota compared to that of humans. We conducted preliminary study provide fundamental information regarding to regional differences in swine gut microbiota. Swine fecal samples were obtained from farms in Jeju (n=40), Gwangju (n=28), and Haenam (n=30). MiSeq was used to sequence 16S rRNA V4 region, and Mothur pipeline (Schloss et al., 2009) was used for data processing. A total of 5,642,125 reads were obtained and 3,868,143 reads were remained after removing erroneous reads. Analysis of taxonomic composition at the phylum level indicated greater abundance of Firmicutes among Jeju swine, and cluster analysis of distribution of operational taxonomic units also showed regional differences among swine gut microbiota. In addition, correlation analysis between non-metric multidimensional scaling and abundance of phyla suggested that the phyla Actinobacter, Verrucomicrobia, Firmicutes, and Fibrobacteres were driving factors for the regional differences. Livestock gut microbiota may be affected by diet and practices in farms. Our results indicated significant regional differences in swine gut microbiota, suggesting that future livestock gut microbiota studies should be designed with the regional differences in mind.