• Title/Summary/Keyword: Protein delivery

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Development of Transgenic Carrot Oral Vaccine to Protect against Diarrhea of Piglets (자돈 설사병 방지를 위한 경구백신용 형질전환 당근 개발)

  • 이영선;황철호
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.287-293
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    • 2002
  • We are trying to develop a transgenic carrot with aims of production and delivery of oral vaccine against microbial enteropathogen using a K88ac pilin gene. A K88ac antigen (pilin) gene was isolated by PCR from the K88ac genomic DNA. The pilin gene was constructed in pGA748 and introduced via Agrobacterium tumefaciens to the explants of carrot hypocotyl and then 494 transgenic lines were established. The amounts of the K88ac antigen produced in each of the cell lines were determined by western and two elite cell lines (M1-17, Y14-1) were selected based on higher levels of expression of the antigens as well as rate of cell growth and efficiency of embryogenesis. In order to test an immunization induced by oral administration of the transgenic carrot, serum of the mice fed with the carrot vaccine were tested in ELISA. It tumed out that the mice fed with 3 g of transgenic carrot showed a similar level of antibody compared to those applied with 10 $\mu\textrm{g}$ of the purified recombinant pilin protein. Besides, various clinical responses were measured after challenging with ETEC K88ac strain to the piglets experiencing an oral immunization with the transgenic carrot. The piglets fed with carrot vaccine showed a lower level of diarrhea in fecal score compared to those fed with non-transgenic carrot. A higher level of increase in weight of the piglets fed with the transgenic carrot vaccine was observed comparing to those fed with non-transgenic carrot as control.

Effect of cultivar and ascorbic acid on in vitro shoot regeneration and development of bombardment-mediated plastid transformation of tomato (Lycopersicon esculentum) (토마토 재분화 효율 향상 및 엽록체 형질전환 조건)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Jong-Bum;Lee, Seung-Bum;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.77-83
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    • 2010
  • Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

Betaine Induces Epidermal Differentiation by Enhancement of Autophagy through an mTOR-independent Pathway (Betaine의 mTOR 비의존적 자가포식 작용 촉진에 의한 표피 분화 유도 효과)

  • Choi, Seon-Guk;Kim, Mi-Sun;Kim, Jin-Hyun;Park, Sun Gyoo;Lee, Cheon Koo;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.1
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    • pp.95-101
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    • 2018
  • The epidermis which is stratified by epithelial tissue renewal based on keratinocyte differentiation protects the organism from various environmental insults by forming a physical barrier. Autophagy is a mechanism which mediates lysosomal delivery and degradation of protein aggregates, damaged organelles and intracellular microorganisms. Recent reports have shown that autophagy has critical roles for proper terminal differentiation to stratum corneum via removing metabolic organelles and nuclei. However, whether increasing autophagy can activate epidermal differentiation is unknown. Here, we screened a library of natural single compounds and discovered that betaine specifically increased the LC3 positive cytosolic punctate vesicles and LC3-I to LC3-II conversion in HaCaT human keratinocyte cell line, indicating increased autophagy flux. mTOR pathway, which negatively regulates autophagy, was not affected by betaine treatment, suggesting betaine-induced autophagy through an mTOR-independent pathway. Betaine-induced autophagy was also observed in primary human keratinocyte and skin equivalent. Furthermore, epidermal thickness was increased in skin equivalent under betaine treatment. Overall, our finding suggests that betaine as a novel regulator of autophagy may induce epidermal turnover and improve the skin barrier abnormality of the aged epidermis.

Synthesis of Thermosensitive and Biodegradable Methoxy Poly(ethylene glycol)-Polycaprolactone and Methoxy Poly(ethylene glycol)-Poly(lactic acid) Block Copolymers (온도감응 및 생분해성 폴리에틸렌 글리콜-폴리카프로락톤과 폴리에틸렌 글리콜-폴리락타이드 공중합체의 합성)

  • 서광수;박종수;김문석;조선행;이해방;강길선
    • Polymer(Korea)
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    • v.28 no.3
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    • pp.211-217
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    • 2004
  • The sol to gel transition of aqueous solution of block copolymers consisting of methoxy poly (ethylene glycol) (MPEG) and biodegradable polyesters such as $\varepsilon$-caprolactone and L-lactide was investigated as a function of temperature. MPEG-PCL was prepared by ring opening polymerization of $\varepsilon$-caprolactone in the presence of HClㆍEt$_2$O as monomer activator at room temperature. Also, MPEG-PLLA was prepared by ring opening polymerization of L-lactide in the presence of stannous octoate at 115$^{\circ}C$. The properties of block copolymers were investigated by $^1$H-NMR, IR, and GPC as well as the observation of thermo sensitive phase transition in aqueous solution. As the hydrophobic block length increased, the sol to gel transition temperature increased and curve of that steepen to lower concentration. To confirm the gel formation at body temperature, we observed the formation of gel in the mice body after injection of 20 wt% aqueous solution of each block copolymer. After surging, we investigated the gelation in mice. The results obtained in this study confirmed the feasibility as biomaterials of injectable implantation for controlled release of drug and protein delivery.

한국농촌의 식품금기에 관한 연구

  • 모수미
    • Journal of the Korean Home Economics Association
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    • v.5 no.1
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    • pp.733-739
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    • 1966
  • A 371 agricultural households from 26 different communities in South Korea was subjected on a study of food taboos in January of 1966. To the pregnant women, those to whom a high protein diet is particurally important, as many as 14 different kinds of foods, mostly portein rich foods, were avoided to eat. It is believed that if duck is eaten while pregnant her baby may walk like a duck in later life. Some mother have a strong aversion to the rabbit meat that her unborn baby must be a harelip. It is feared to eat chicken, shark or carp by the pregnant mother for her baby may get a gooseflesh appearance, or fish scale-like skin in later life. It is thought that if mother eats soup made of meat borns, especially chicken bones, a disfigured baby may be born. Some area informed that if mother eats crab meat her future baby will always bubble. To the child-bearing mothers 13 different kinds of foods were avoided to eat. Some believe that if raddish kimchi, soybean curd, squash are eaten while dilivery that mother may get dental decay or to lose all her teeth. Other think that highly spiced raddish kimchi cause delivery difficult. To the lactating mothers 7 different items of foods were not recommended to eat. It is a common belief that eating green vegetables, especially fresh lettuce, are restricted that her baby may stool greenish. It is said that eating ginsen-chicken soup, or ginsen tea during lactating reduces breast milk secretion. To the weaning babies 7 different kinds of foods were prohibited to fee. Eggs are not eaten because mothers think her babies will start to talk very late. Eight different items of foods in cases of gastro-intestinal diseases, 5 items for liver disease, 7 items for high blood pressure as well as for paralysis were respectively restricted. It is said that meats including pork, beef, and chicken are neither desirable for the patients of high blood pressure nor those of paralysis. To the measles children 10 varieties of foods were restricted. Especially soybean products and meats were not encouraged to use for avoiding asecond attack of measles. For the common cold 8 different kinds of foods were aversed and men think that eating of soup of undria delays a recovery. For the tuberculosis 4 kinds of foods were prohibited to eat. It is said that wine, red pepper and ginsen will stimulate lung bleeding. Many mothers had a strong aversion to fermented shrimp and fish in case of style. and 5 different items of foods were restricted. In case of menstration not so many foods were restricted as other cases, but meat soup is not eaten in this condition in some areas. Majority of food taboos in Korean villages are neither based on tribal nor religious factors. But no one knows how, since what ages, from where, these food taboos have been transmitted and spread over the country. This survey found a great variety of food taboos, aversions, traditional beliefs and prohibitions latent unknown reseasons, or non-scientific conceptions, or completely different ideas from the modern medical aspect, or somewhat fallacious and superstitious beliefs. For the vascular disease contrasting approach were found between modern the oritical therapy and popular remedy among the rural populations who largely depend on the eastern medication. Further scientific study on either side should be done to lead the patient proper way. Many restricted foods such as rabbit, duck, chicken and fish are best resources of protein rich foods which are available in the village. Emphasis should be laid upon breaking down fallacious and supersititious food taboos through the extended nutrition education activities in order to improve food habit and good eating pattern for healthier and stronger generations of Korea.

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Effect of Subtotal Pancreatectomy of Female Mice on the Embryo and Offspring (암컷 생쥐의 췌장 부분절제가 배아와 신생자에 미치는 영향)

  • Cho, Young-Jae;Lee, Jung-Sun;Gu, Sun-Kyung;Park, Moon-Nyeo;Lee, Ha-Kye;Cho, Byung-Nam
    • Development and Reproduction
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    • v.10 no.3
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    • pp.185-192
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    • 2006
  • It is known that maternal diabetes induces a premature delivery and a congenital malformation. Although it was suggested that hyperglycemia is a main cause, little has been known. The effects of subtotal pancreactomized animal as a diabetic model on the embryo and the offspring were investigated. 1) Although survival rate of offsprings from the 90% pancreactomized female was lower than that of control, body weight of the survived offsprings was similar to that of control until 6 weeks after birth. When the liver was investigated, abnormal tissue was observed in both female and male offsprings from the pancreactomized mother. The level of major urinary protein(MUP) that is synthesized in the liver and secrets into urine was decreased in both female and male, supporting the liver abnormality. 2) In the pancreactomized female, size of the Langhan's island was increased and the liver also was observed abnormal. However, blood glucose level of the pancreactomized mother was not different from that of control. The low body weight and survival rate of offsprings from the $inhibin-{\alpha}$ overexpressed mice was very similar to those of the offsprings from diabetic mother. Thus when the mother itself was investigated after DNA injection, blood glucose level in the $inhibin-{\alpha}$ overexpressed mice was not different from that of control although size of the Langhan's island was decreased. Taken together, subtotal pancreactomy affected the offspring and embryo and influenced deeply the mother itself. However glucose level of the mother was not changed during investigation. It suggested that the relationship between the diabetic mother and embryo through offspring is very complex.

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Comparison of Blood and Urine Renal Indices Between Hypercalciuric and Non-hypercalciuric Hematuria Patients (혈뇨 환아에서 고칼슘뇨군과 비고칼슘뇨군의 혈액 및 소변화학검사와 신기능 지표들의 비교)

  • Lee, Jin-Hee;Lee, Hyun-Seung;Lee, Keun-Young;Jang, Pil-Sang;Lee, Kyung-Yil;Kim, Dong-Un
    • Childhood Kidney Diseases
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    • v.11 no.2
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    • pp.168-177
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    • 2007
  • Purpose : The purpose of this study was to investigate whether hypercalciuria patients with hematuria show different renal indices compared to non-hypercalciuria patients with hematuria. Methods : We retrospectively reviewed the medical records of patients with gross or microscopic hematuria whose blood chemistry and 24 hour urine chemistry were examined. After excluding the patients with more than $4 mg/m^2/day$ proteinuria or the patients with urinary calcium excretion between 3 and 4 mg/kg/day, we divided the patients into two groups: a hypercalciuria group whose calcium excretion was more than 4 mg/kg/day(n=30) and a non hypercalciuria group whose calcium excretion was less than 3 mg/kg/day(n=41). The urinary excretion, clearance, and fractional excretion(FE) of Na, K, Cl, Ca, P, urea, and creatinine were calculated and compared between the two groups. Results : The hypercalciuria group had more calcium excretion($6.1{\pm}2.9$ vs $1.5{\pm}0.9 mg/kg/day$), more urea excretion($341{\pm}102$ vs $233{\pm}123 mg/kg/day$), greater glomerular filtration rate(GFR) ($93.7{\pm}31.1$ vs $79.5{\pm}32.0 mL/min$) but lower FENa($1.0{\pm}0.4%$ vs $1.3{\pm}0.6%$) than the nonhyper-calciuria group, although the urinary sodium excretion was similar between the two groups. Conclusion : The greater urea excretion and GFR in hypercalciuric patients suggest that they might be on a higher protein diet than the non-hypercalciuria group. The increased glomerular filtration of sodium and calcium induced by the higher GFR in hypercalciuria would have increased their delivery to the distal tubule, where sodium is effectively reabsorbed but calcium is not, which is suggested by the lower FENa but higher FECa in hyercalciuria. It is recommended that the diet of hematuria patients be reviewed in detail at initial presentation and during treatment.

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A prognosis discovering lethal-related genes in plants for target identification and inhibitor design (식물 치사관련 유전자를 이용하는 신규 제초제 작용점 탐색 및 조절물질 개발동향)

  • Hwang, I.T.;Lee, D.H.;Choi, J.S.;Kim, T.J.;Kim, B.T.;Park, Y.S.;Cho, K.Y.
    • The Korean Journal of Pesticide Science
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    • v.5 no.3
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    • pp.1-11
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    • 2001
  • New technologies will have a large impact on the discovery of new herbicide site of action. Genomics, combinatorial chemistry, and bioinformatics help take advantage of serendipity through tile sequencing of huge numbers of genes or the synthesis of large numbers of chemical compounds. There are approximately $10^{30}\;to\;10^{50}$ possible molecules in molecular space of which only a fraction have been synthesized. Combining this potential with having access to 50,000 plant genes in the future elevates tile probability of discovering flew herbicidal site of actions. If 0.1, 1.0 or 10% of total genes in a typical plant are valid for herbicide target, a plant with 50,000 genes would provide about 50, 500, and 5,000 targets, respectively. However, only 11 herbicide targets have been identified and commercialized. The successful design of novel herbicides depends on careful consideration of a number of factors including target enzyme selections and validations, inhibitor designs, and the metabolic fates. Biochemical information can be used to identify enzymes which produce lethal phenotypes. The identification of a lethal target site is an important step to this approach. An examination of the characteristics of known targets provides of crucial insight as to the definition of a lethal target. Recently, antisense RNA suppression of an enzyme translation has been used to determine the genes required for toxicity and offers a strategy for identifying lethal target sites. After the identification of a lethal target, detailed knowledge such as the enzyme kinetics and the protein structure may be used to design potent inhibitors. Various types of inhibitors may be designed for a given enzyme. Strategies for the selection of new enzyme targets giving the desired physiological response upon partial inhibition include identification of chemical leads, lethal mutants and the use of antisense technology. Enzyme inhibitors having agrochemical utility can be categorized into six major groups: ground-state analogues, group specific reagents, affinity labels, suicide substrates, reaction intermediate analogues, and extraneous site inhibitors. In this review, examples of each category, and their advantages and disadvantages, will be discussed. The target identification and construction of a potent inhibitor, in itself, may not lead to develop an effective herbicide. The desired in vivo activity, uptake and translocation, and metabolism of the inhibitor should be studied in detail to assess the full potential of the target. Strategies for delivery of the compound to the target enzyme and avoidance of premature detoxification may include a proherbicidal approach, especially when inhibitors are highly charged or when selective detoxification or activation can be exploited. Utilization of differences in detoxification or activation between weeds and crops may lead to enhance selectivity. Without a full appreciation of each of these facets of herbicide design, the chances for success with the target or enzyme-driven approach are reduced.

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Maternal Vitamin $B_{6}$ Intake and Vitamin $B_{6}$ Level in Maternal, Umbilical Cord Plasma and Placenta (임신부의 비타민 $B_{6}$ 섭취와 모체와 제대혈 및 태반 조직의 비타민 $B_{6}$농도)

  • 안홍석;이금주;정환욱
    • Journal of Nutrition and Health
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    • v.35 no.3
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    • pp.322-331
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    • 2002
  • This study was carried out to evaluate the effect of vitamin $B_{6}$ intake by normal term delivery pregnant women on the concentration of vitamin $B_{6}$ in the material plasma, the umbilical cord plasma, and the placental tissue. Dietary intake data were obtained from a semi-quantitative frequency questionnaire. The daily mean energy and protein intakes were 2189.5 kcal (93.2% of RDA) and 79.3 g (113.3% of RDA), respectively. The average daily vitamin $B_{6}$ intake was 1.7 mg (91.4% of RDA) for the pregnant women. Their main sources of vitamin $B_{6}$ were cereal & starch (50%), and vegetables & fruits (33%). The pyridoxal phosphate (PLP) concentration of the maternal plasma, the umbilical cord plasma, and the placenta were 16.7 $\pm$ 4.1 nmol/1, 61.3 $\pm$ 19.8 nmol/l and 898.6 $\pm$ 159.2 ng/g, respectively. The PLP level was the highest in the placenta. The PLP level of the maternal plasma was significantly lower than the of the umbilical cord plasma (p < 0.001). The PLP level of maternal plasma correlated positively with that of the placenta (p < 0.0001) and the umbilical cord plasma (p < 0.05). Also the PLP level of the placenta correlated positively with that of the umbilical cord plasma (p < 0.05). These findings indicate that the vitamin $B_{6}$ nutritional status of the fetus is affected by placental vitamin $B_{6}$ levels, and that the placental vitamin B$_{6}$ levels reflect the maternal vitamin $B_{6}$ status. The umbilical cord plasma PLP level showed a positive correlation with the gestational length (p < 0.05). A negative association was observed between the PLP level showed of the umbilical cord plasma and the pregnancy weight gain (p < 0.03). The results suggest that the transfer of PLP from maternal plasma to the placental tissue could be an active transport, white the transfer of PLP from the placenta to the fetus is by means of simple diffusion. Thus, neonatal vitamin $B_{6}$ nutrition is influenced by the maternal nutritional status.

Production of Medium-chain Fatty Acids in Brassica napus by Biotechnology (유채에서의 중쇄지방산 생산)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Hyun-Uk;Lee, Kyeong-Ryeol;Kim, Jong-Bum
    • Journal of Applied Biological Chemistry
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    • v.53 no.2
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    • pp.65-70
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    • 2010
  • Medium-chain fatty acids (MCFA) are composed of 8-12 carbon atoms, and are found in coconut, cuphea, and palm kernel oil. MCFA were introduced into clinical nutrition in the 1950s for dietary treatment of malabsorption syndromes because of their rapid absorption and solubility. Recently, MCFA have been applied to Gastrointestinal Permeation Enhancement Technology (GIPET), which is one of the most important parts in drug delivery system in therapeutics. Therefore, to accumulate the MCFA in seed oil of rapeseed, much effort has been conducted by classical or molecular breeding. Laurate can be successfully accumulated up to 60 mol% in the seed oil of rapeseed by the expression of bay thioesterase (Uc FatB1) alone or crossed with a line over-expressing the coconut lysophosphatidic acid acyltransferase (LPAAT) under the control of a napin seed-storage protein promoter. Also, caprylate and caprate were obtained 7 mol% and 29 mol%, respectively, from plants over-expressing of the medium-chain specific thioesterase (Ch FatB2) alone or together with the chain-length-specific condensing enzyme (Ch KASIV). Despite the success of some research in utilizing parallel classical and molecular breeding to produce MCFA, commercially available seed oils have for the most part, not been realized. Recent research in the field of developing MCFA-enriched transgenic plants has established that there is no single rate-limiting step in the production of the target fatty acids. The purpose of this article is to review some of the recent progress in understanding the mechanism and regulation of MCFA production in seed oil of rapeseed.