• 제목/요약/키워드: addition-chain

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Variations of Properties and Microbial Community during Fermentation of Makgeollies by Isolated Yeasts from Traditional Makgeollies (전통막걸리에서 분리한 효모균주를 이용한 막걸리 발효과정 중의 물성 및 미생물 군집의 변화)

  • Jeon, Myong Je;Jang, Min Kyung;Lee, Sol Jee;Park, Sung Hwan;Kim, Mihyang;Sohn, Jae Hak;Lee, Han-Seung;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • 제23권6호
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    • pp.796-803
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    • 2013
  • Property changes and bacterial characterizations by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) were investigated during the fermentation of Makgeollies by 5 isolated yeast strains. Changes of pH were large between day 0 (pH 6) and day 2 (pH 3) and showed less variation after then. ANOVA analyses revealed that pHs were statistically different with fermentation times (p<0.001), while strains (p=0.60) did not. Acidities were changed from 0.19 to 1.04% and showed rather high increase from day 2, and fermentation times (p<0.001) and strains (p=0.006) represented statistical differences. All strains showed less than 0.150% at amino-type nitrogen contents except S strain showed 0.442% at day 8, and there were no statistical differences with fermentation times (p=0.4558) and strains (p=0.3513). Saccharinities of C strain were higher from day 4, and fermentation times (p<0.0001) and strains (p=0.007) showed statistical differences. Large variation of alcohol concentrations (%) were observed between day 0 (0%) and day 2 (10%) and showed less variation after day 2, and there was no statistical difference with strains. Dominant prokaryotes were Lactobacillus fermentum and Pediococcus pentosaceus, which producing acids and functional materials. Dominant eukaryote was Saccharomyces cerevisiae, which might be resulted from addition of yeasts.

Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2014년도 춘계학술대회 및 임시총회
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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Trends and Prospects of N. Korea Military Provocations After the Sinking of ROKS Cheon-an (천안함 폭침 이후 북한의 군사도발 양상과 전망)

  • Kim, Sung-Man
    • Strategy21
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    • 통권34호
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    • pp.58-92
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    • 2014
  • Even after S. Korea took 5.24 Measure(24 May 2014), N. Korea has not stopped raising provocations such as the shelling of Yeonpyeong Island, electronic and cyber attacks. To make matters worse, the communist country lunched long-range missiles(twice) and conducted 3rd nuclear test, escalating tensions which could possibly lead to an all-out war. Korean Government failed to respond properly. However, escalation into an all-out war was deterred by the CFC immediately carrying out its peacetime duty(CODA). The US made a rapid dispatch of its augmentation forces(Aircraft carrier, nuclear-powered submarine, strategic bomber, F-22) to the Korean Peninsula. In recognition of the importance of the Combined Forces Command, since May 2013 the Park Geun-Hye Administration has been pushing ahead with re-postponement of Wartime Operational Control Transfer(which initially meant the disassembling of the CFC as of 1 December 2015) More recently, there has been a series of unusual indicators from the North. Judging from its inventory of 20 nuclear weapons, 1,000 ballistic missiles and biochemical weapons, it is safe to say that N. Korea has gained at least war deterrence against S. Korea. Normally a nation with nuclear weapons shrink its size of conventional forces, but the North is pursuing the opposite, rather increasing them. In addition, there was a change of war plan by N. Korea in 2010, changing 'Conquering the Korean Peninsula' to 'Negotiation after the seizure of the Greater Seoul Metropolitan Area(GSMA)' and establishing detailed plans for wartime projects. The change reflects the chain reaction in which requests from pro-north groups within the South will lead to the proclamation of war. Kim, Jeong-Un, leader of N. Korean regime, sent threatening messages using words such as 'exercising a nuclear preemptive strike right' and 'burning of Seoul'. Nam, Jae-June, Director of National Intelligence Service, stated that Kim, Jung-Un is throwing big talks, saying communization of the entire Korean Peninsula will come within the time frame of 3 years. Kim, Gwan-Jin, Defense Minister, shared an alarming message that there is a high possibility that the North will raise local provocations or a full-fledged war whenever while putting much emphasis on defense posture. As for the response concept of the Korean Government, it has been decided that 'ROK·US Combined Local Provocation Counter-Measure' will be adopted to act against local provocations from the North. Major provocation types include ▲ violation of the Northern Limit Line(NLL) with mobilization of military ships ▲ artillery provocations on Northwestern Islands ▲ low altitude airborne intrusion ▲ rear infiltration of SOF ▲ local conflicts within the Military Demarcation Line(MDL) ▲ attacking friendly ships by submarines. Counter-measures currently established by the US involves the support from USFK and USFJ. In order to keep the sworn promise, the US is reinforcing both USFK and USFJ. An all-out war situation will be met by 'CFC OPLAN5027' and 'Tailored Expansion Deterrence Forces' with the CFC playing a central role. The US augmentation forces stands at 690,000 troops, some 160 ships, 2,000 aircraft and this comprise 50% of US total forces, which is estimated to be ninefold of Korean forces. The CFC needs to be in center in handling both local provocations and an all-out war situation. However, the combat power of S. Korean conventional forces is approximately around 80% of that of N. Korea, which has been confirmed from comments made by Kim, Gwan-Jin, Defense Minister, during an interpellation session at the National Assembly. This means that S. Korean forces are not much growing. In particular, asymmetric capabilities of the North is posing a serious threat to the South including WMD, cyber warfare forces, SOF, forces targeting 5 Northwestern Islands, sub-surface and amphibious assault forces. The presence of such threats urgently requires immediate complementary efforts. For complementary efforts, the Korean Government should consider ① reinforcement of Korean forces; putting a stoppage to shrinking military, acquisition of adequate defense budget, building a missile defense and military leadership structure validity review, ② implementation of military tasks against the North; disciplinary measures on the sinking of ROKS Cheon-an/shelling of Yeonpyeong Islands, arrangement of inter-Korean military agreements, drawing lessons from studies on the correlation between aid for N. Korea, execution of inter-Korean Summit and provocations from the North, and ③ bolstering the ROK·US alliance; disregarding wartime operational control transfer plan(disassembling of CFC) and creation of a combined division.

Consumer Perceptions of Food-Related Hazards and Correlates of Degree of Concerns about Food (주부의 식품안전에 대한 인식과 안전성우려의 관련 요인)

  • Choe, Jeong-Sook;Chun, Hye-Kyung;Hwang, Dae-Yong;Nam, Hee-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • 제34권1호
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    • pp.66-74
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    • 2005
  • This survey was conducted to assess the consumer perceptions of food-related hazard in 500 housewives from all over Korea. The subjects were selected by stratified random sampling method. The survey was performed using structured questionnaire through telephone interview by skilled interviewers. The results showed that 34.6% of the respondents felt secure and were not concerned about food safety, and 65.4% were concerned about food safety. Logistic regression analysis showed that the increasing concern on food brands, food additives (such as food preservatives and artificial color), and imported foodstuffs indicated the current increasing concern on food safety. Other related factors indicating the increasing concern on food safety were education level and care for children's health. The respondents who cared about food safety expressed a high degree of concern on processed foodstuffs such as commercial boxed lunch (93.3%), imported foods (92.7%), fastfoods (89.9%), processed meat products (88.7%), dining out (85.6%), cannery and frozen foods (83.5%), and instant foods (82.0%). The lowest degree of concern was on rice. All the respondents perceived that residues of chemical substances such as pesticides and food additives, and endocrine disrupters were the most potential food risk factors, followed by food-borne pathogens, and GMOs (Genetically Modified Organisms). However, these results were not consistent with scientific judgment. Therefore, more education and information were needed for consumers' awareness of facts and myths about food safety. In addition, the results showed that consumers put lower trust in food products information such as food labels, cultivation methods (organic or not), quality labels, and the place of origin. Nevertheless, the respondents expressed their desire to overcome alienation, and recognized the importance of knowing of the origin or the producers of food. They identified that people who need to take extreme precautions on food contamination were the producers, government officials, food companies, consumers, the consumer's association, and marketers, arranged in the order of highest to lowest. They also believed that the production stage of agriculture was the most important step for improving the level of food safety Therefore, the results indicated that there is a need to introduce safety systems in the production of agricultural products, as follows: Good Agricultural Practice (GAP), Hazard Analysis and Critical Control Point (HACCP), and Traceability System (75).

PERIODONTOPATHIC BACTERIA AND ANTIBIOTIC RESISTANCE GENES OF ORAL BIOFILMS IN CHILDREN (어린이 치면세균막에서 치주질환원인균과 항생제 내성유전자의 출현율)

  • Kim, Seon-Mi;Choi, Nam-Ki;Cho, Seong-Hoon;Lee, Seok-Woo;Lim, Hoi-Jeong;Lim, Hoi-Soon;Kang, Mi-Sun;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • 제38권2호
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    • pp.170-178
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    • 2011
  • The purpose of this study was to assess the prevalence of periodontopathic bacteria and resistance determinants from oral biofilm of children. Subgingival dental plaque was isolated from 87 healthy children, and PCR was performed to determine the presence of 5 periodontal pathogens including P. gingivalis, T. forsythia, T. denticola, F. nucleatum, A. actinomycetemcomitans, and nine resistance genes including tet(Q), tet(M), ermF, aacA-aphD, cfxA, $bla_{SHV}$, $bla_{TEM}$, vanA, mecA. 1. The prevalence of F. nucleatum, T. forsythia. and P. gingivalis was 95.4%, 55.2%, and 40.2%, respectively. In addition. the prevalence of A. actinomycetemc omitans was 5.7%, while T. denticola was 3.4%. 2. In analysis of antibiotic resistance determinants. cfxA, $bla_{TEM}$ and tet(M) were detected in all the samples tested. It was also found that the prevalence of tet(Q) showing tetracycline resistance. $bla_{SHV}$ associated with resistance to ${\beta}$-lactams, ermF exhibiting erythromycin resistance, and, vanA resulting vancomycin resistance was 88.5%, 29.9% 87.4%, and 48.5%, respectively. The aacA-aphD gene showing resistance to aminoglycosides and mecA gene harboring methicillin resistance exhibited the lowest prevalence with 9.2%. 3. In a correlation analysis between periodontopathic pathogens and antibiotic resistance determinants, it was found that there was a significant correlation between T. forsythia and $bla_{SHV}$. Also, P. gingivalis and vanA showed a correlation. Finally, tet(Q) and ermF showed a significant correlation (phi: 0.514) while mecA and vanA also showed a correlation(phi: 0.25).

Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • 제26권3호
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

Toll-like Receptor 4-mediated Apoptotic Cell Death in Primary Isolated Human Cervical Cancers (부인과질환 특이적 종양의 TLR4 매개성 apoptosis 유발에 관한 연구)

  • Won, Jinyoung;Hong, Yunkyung;Park, Sookyoung;Kim, Joo-Heon;Hong, Yonggeun
    • Journal of Life Science
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    • 제28권6호
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    • pp.718-725
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    • 2018
  • Toll-like receptor 4 (TLR4) has been implicated in cell proliferation and apoptosis in several types of cancer. In this study, the impact of TLR4 activation on apoptotic cell death in gynecologic cancers induced by lipopolysaccharide (LPS) was investigated. Cervical cancer cell lines were produced from isolated surgical specimens supplied by Paik Hospital. The primary cultures of normal myometrium and gynecologic cancers, including cervical, endometrial, and ovarian cancers, were used to examine the differences in morphological characteristics between normal and cancerous cells. A reverse transcription polymerase chain reaction analysis was used to determine the relative expression levels of TLR4 gene involved in apoptosis-associated signaling in cervical cancer cells. The cancer cell colonies showed a tendency to reach high levels of confluency compared with normal cells. In addition, an enhanced growth rate and loss of contact inhibition were observed in gynecologic cancer cells compared with normal cells (doubling times of 16.6 hr vs. 26 hr, respectively). The expression level of ITGA5, an alpha-5 integrin marker, was upregulated in normal myometrial cells, but this tendency was not exhibited in cervical cancer cells. Furthermore, p53 tumor suppressor gene expression was upregulated, whereas TLR4 and caspase-3 gene expressions were downregulated in cervical cancer cells. Notably, the expression levels of TLR4 and caspase-3 were increased significantly in LPS-treated cancer cells compared with those in non-LPS-treated cells. These results suggest that the TLR4-mediated caspase-dependent apoptotic signaling pathway could be suggested as a therapeutic target for the treatment of gynecologic cancers, including cervical cancers.

Effects of Added Vegetable Oils on In vitro Formation of Fatty Acid Soaps and Fermentation Characteristics and NDF Disappearance Rate (식물성유 첨가가 In vitro 발효성상, NDF 소실율 및 지방산염 형성에 미치는 영향)

  • Kim, D.I.;Choi, J.R.;Lee, Y.H.;Lee, J.K.;Chung, T.Y.
    • Journal of Animal Science and Technology
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    • 제46권3호
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    • pp.355-372
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    • 2004
  • In vitro experiments were conducted to determine the formation of fatty acid soaps (FAS) and neutral detergent fiber (NDF) disappearance rate. The substrates were a basal alfalfa hay containing 1) no oil, 2) 10% soybean oil, 3) 10% com oil, on a weight basis. All the substrates were incubated in triplicate for 0, 3, 6, 12, 24 and 48h in each experiment. After the incubation in the first experiment serum bottles (6oml) were analyzed for nonesterified, esterifed and fatty acid soaps contents. The serum bottles (120mI) from the second experiment were analyzed for pH, $NH_3-N$ and VFA concentration, and dry matter and NDF disappearance rate. pH decreased and the concentration of NH3-N increased significantly with longer incubation time (P<0.0001). The disappearance rates of dry matter and NDF significantly varied with feed, incubation time and oils (P<0.05). The molar concentration of total VFA increased and proportion of acetate significantly decreased with incubation time (P<0.0001), but the proportion of propionate significantly increased with longer incubation time (P<0.0001). Addition of oils to diet lowered the ratio of acetate:propionate (P<0.05). The esterified fatty acids (EFA) decreased with increasing incubation time (P<0.0001), and nonesterified fatty acids (NEFA) increased due to lipolysis of EFA, NEFA then reacted with cations to increase formation of FAS. The formation of FAS increased significantly at 48h of incubation time (P<0.0001). Especially, formation of stearic acid soaps was 27.5 and 32.5 folds with soybean oil and com oil supplements, respectively, by 48h of incubation time (P<0.0001). Alfalfa hay had higher cation contents, particularly Ca, which react with NEFA and FAS can be formed with longer incubation time. Saturated fatty acids had a higher proportion of FAS than did unsaturated fatty acids, suggesting that the former may react more extensively with cations. FAS contents increased with increasing chain length of the fatty acids. Since added vegetable oils fonned FAS, it might decrease negative effects on in vitro fermentation characteristics and NDF disappearance rate.

Structural and emulsification properties of octenyl succinylated potato dextrin upon different preparation methods (OSA-감자 덱스트린의 구조 및 유화 특성 연구)

  • Han, Yu-Jin;Li, Shun Ji;Han, Jung-Ah
    • Korean Journal of Food Science and Technology
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    • 제49권1호
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    • pp.8-13
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    • 2017
  • Octenyl succinylated (OSA) potato starch was dextrinized by two methods: ultrasound (at 25, 50, or $70^{\circ}C$ for 1 h; OSA-25UT, OSA-50UT, and OSA-70UT, respectively) and acid hydrolysis (for 1 or 4 h; OSA-AD1H or OSA-AD4H, respectively), and the properties of the resulting starch were analyzed. The melting enthalpy of OSA-70UT decreased the most (from 14.0 to 10.0 mJ/mg), indicating chain degradation. For pasting properties, as ultrasound treatment temperature increased, peak viscosity decreased (2884, 2550, and 1888 cP, respectively), whereas acid hydrolysis increased peak viscosity and decreased pasting temperature. The relative crystallinity of OSA-dextrin produced by ultrasound or acid hydrolysis significantly decreased (from 33.61 to 14.90-26.03 and 19.28-20.05, respectively) as temperature or time increased, yet a B-type crystal pattern was maintained. Regarding emulsifying stability and sensory tests of mayonnaise prepared with OSA potato dextrin, mayonnaise with OSA-70UT was stable for short storage period (1 week), however mayonnaise with OSA-AD1H was the most suitable for long storage periods (from 2 to 4 weeks). In addition, the OSA-70UT was the most acceptable for mayonnaise in the sensory test.

The Separate and Combined Effects of Insulin, Dexamethasone and Growth Hormone on the OB Gene Expression and Leptin Secretion from Cultured Human Visceral Adipose Tissue (인체의 복강 내 지방조직 배양을 통한 OB 유전자 발현과 Leptin 분비에 미치는 인슐린, Dexamethasone과 성장호르몬의 단독 또는 복합적 영향에 관한 연구)

  • Hwang, Il Tae;Kim, Kyung Hee;Hwang, Jin Soon;Shin, Choong Ho;Yang, Sei Won
    • Clinical and Experimental Pediatrics
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    • 제46권8호
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    • pp.795-802
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    • 2003
  • Purpose : We investigated the hormonal control of OB gene expression and leptin secretion in cultured human visceral adipose tissue. Methods : Visceral adipose tissues were cultured for up to 48 hrs in modified Eagle's medium with varying concentration of hormones : Control(no hormone), bovine insulin(100 nM), Dexamethasone(DEX, 100 nM), growth hormone(GH, 40 ng/mL), insulin+DEX(100 nM each), insulin+DEX+GH(100 nM insulin and DEX, 40 ng/mL GH). Quantitative analysis of leptin mRNA was performed by competitive reverse transcription polymerase chain reaction, and leptin secretion in culture medium was measured by IRMA using a commercial kit. Results : The addition of dexamethasone to the medium significantly increased OB gene expression and leptin secretion(P<0.05). Unlike dexamethasone, insulin did not affect OB gene expression and leptin secretion. Both insulin and dexamethasone, at high concentration, significantly stimulated leptin secretion compared with basal values(P<0.05). Leptin gene expression was not significantly increased by GH treatment alone, however GH, in combination with high concentrations of insulin and dexamethasone, attenuated the stimulatory effects of high concentrations of insulin and dexamethasone. Conclusion : Insulin cannot increase leptin secretion without the presence of dexamethasone. The mechanism suggested is that insulin may increase leptin secretion in cytoplasm only after dexamethasone increases the expression of OB gene. Further studies are necessary to elucidate the mechanism of the action of insulin on leptin secretion after increasing OB gene expression by dexamethasone.