• Journal of Life Science
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• v.26 no.3
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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.

• Journal of Animal Science and Technology
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• v.51 no.3
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• pp.207-216
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• 2009

This experiment was conducted to investigate the effects of dietary supplementation of organic acid mixture on growth performance, cecal microflora, blood characteristics and immune response in broiler chicks and to prove the possibility of organic acid mixture as an alternative to antibiotics growth promotor. A total of four hundred eighty, 1-day-old male broiler chicks (Ross$\times$Ross 308) were randomly divided into 4 groups with 4 replicates of 30 birds each. The treatments were NC (free antibiotics), PC (basal diet with virginiamycin 10 ppm and salinomycin 60 ppm), 0.3% organic acid, and 0.5% organic acid. The final body weight and body weight gain were significantly higher in organic acid 0.5% than NC (P<0.05). The feed conversion ratio in all treated groups were significantly improved as compared to that of NC (P<0.05). The carcass rate and relative organs weight were not significantly difference among the groups. The relative weight and length of small intestine in PC were significantly decreased than the other groups. The numbers of cecal coliform bacteria and Salmonella in all treated groups were significantly lower than NC (P<0.05). The number of cecal lactic acid bacteria was not different among the groups. No significant differences among the groups were observed in the contents of total cholesterol, triglyceride, blood urea nitrogen (BUN), albumin, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in blood serum. The contents of total protein and globulin in blood serum of PC and organic acid treated groups were significantly increased as compared to those of NC (P<0.05). Therefore, albumin:globulin ratio of PC and organic acid treated groups was significantly lower than NC (P<0.05). The total white blood cell (WBC), heterophil, lymphocyte, and stress indicator (heterophil:lymphocyte ratio) were not significantly different among the groups. No significant difference was observed on the expression rate of splenic cytokines mRNA in organic acid treated groups compared to the control. Consequently, supplemental organic acid mixture improved the growth performance, and influenced positive effects on the intestinal microflora by inhibiting the growth of harmful bacteria without any adverse effects on relative weights of organs and blood biochemical parameters in broiler chicks.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.24-29
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• 2013

In the present study, the effects of prebiotics and prebiotics+probiotics on intestinal microflora and fermentation products were evaluated in a pig in vitro fermentation model. The substrates used in this study were iso-malto oligosaccharide (IMO), partially digested chicory-inulin (CI), raffinose (RA), and cyclodextrin (CD) as prebiotics and Lactobacillus reiteri as probiotics. For a pig in vitro fermentation, the experimental diet for growing pigs was predigested using digestive enzymes secreted by small intestine and this hydrolyzed diet was mixed with a buffer solution containing 5% fresh swine feces. The mixture was then incubated with either prebiotics or prebiotics+probiotics for 24 h. Samples were taken at 24 h, and viable counts of microflora, gas, pH, volatile organic compounds (VOCs) and short-chain fatty acid (SCFA) were analyzed. The viable count of Enterobacteriaceae was significantly decreased (p<0.001) in all treatments containing prebiotics and prebiotics+probiotics when compared to the control. However, the number of lactic acid bacteria increased in the prebiotics and prebiotics+probiotics treatment. The pH values in the fermentation fluid decreased in all treatments when compared to the control, and their effects were greater in the prebiotics+probiotics group than prebiotics group. Fermentation with prebiotics resulted in a reduction in malodorous compounds such as ammonia, hydrogen sulfide and skatole when compared to the prebiotics+probiotics group. Short-chain fatty acid production was also higher for treatment with prebiotics+probiotics than treatment with prebiotics. In conclusion, the results of this study demonstrated that fermentation with prebiotics was effective in reducing the formation of malodorous compounds and prebiotics+probiotics was effective in increasing lactic acid bacteria and SCFA and reducing the pH. Moreover, further studies will be needed to determine whether the results observed in the in vitro model would occur in pigs that ingest these prebiotics or probiotics.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.2
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• pp.186-192
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• 1997

Increased activities of phase 2 enzymes including quinone reductase(QR) have been reported to be associated with protection of animals from neoplastic, mutagenic, and other toxic effects of many carcinogens. In previous study, we found that methanol extract of roasted and defatted perilla meal induced the activity of quinone reductase, an anticarcinogenic marker enzyme, in murine hepalc1c7 cells. Current study showed that unroasted perilla had a limited QR-inducing activity, suggesting that roasting cause the generation of active component(s). Thus we hypothesized that QR inducer in perilla might be covalently linked to sugar moiety and released during roasting process. Methanol extract of defatted raw perilla was subject to acid treatment in order to hydrolyze the potential sugar moiety. Prolonged hydrolysis of methanol extract of defatted raw perilla at $98{\sim}100^{\circ}C$ increased the ability to induce cytosolic QR activity of hepalclc7 cells. Furthermore roasting at 180 and $200^{\circ}C$ resulted in significant induction of QR activity. The result strongly support the idea that QR inducer(s) is present in bound form in raw perilla and released during roasting. Cellular QR activity was induced proportionately with the increase of concentration of methanol extract of roasted perilla. The induction of QR by defatted perilla was also examined in the cytosols of liver, small intestine, stomach, lung and kidney of male ICR mice. Induction patterns showed specificity with respect to target tissue and roasting of perilla. Unroasted perilla meal (defatted) significantly induced QR in liver and lung, while roasted perilla meal induced QR in liver and stomach. The observation that raw perilla showed similar QR induction patterns to roasted perilla is consistent with our proposal that QR inducer(s) is present in bound form and released by physical and chemical treatments as digestive or microbial enzymes could release the inducers from inactive glycoside forms in gastrointestinal tract of mice. In conclusion, perilla could exert protective effect against chemically induced carcinogenesis by inducing phase 2 enzymes in biological systems regardless of chemical and physical process such as roasting.

• Korean Journal of Organic Agriculture
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• v.18 no.3
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• pp.347-361
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• 2010

This study was carried out to investigate the effects of wormwood (Artemisia sp.) addition on the growth performance, nutrients utilization and abdominal fat deposition of broiler chickens. Two hundred twenty five and two-day old Arboracre strain male commercial broiler chicks were distributed to 5 treatments with wormwood supplementation levels; C(O%), $T_1$ (1%), $T_2$ (3%), $T_3$ (5%), and $T_4$ (10%) and with 3 replications each with 5 birds for five weeks. Body weight gain during the experiment was improved in $T_1$ (1723.0g) compared with that of $T_3$ (1557.7g) and $T_4$ (1450.7g) (P<0.05). Feed intake was significantly (P<0.05) increased as the levels of wormwood addition increased (C: 2653.8g, T1: 2852.0, $T_2$: 2900.3, $T_3$: 2900.7g, T4: 2954.7g). Feed conversion rate (feed/gain) was significantly (P<0.05) increased as the levels of wormwood addition increased (C: 1.55, $T_1$: 1.66, $T_2$: 1.70, $T_3$: 1.86, $T_4$: 2.04). The days reaching to 2.0kg of body weight were expected to be 43.2 days in control, whereas those of group T1 were reduced to 42.6 days by 0.6 day from control. Nutrient utilization and abdominal fat deposition in the experiment were significantly decreased (P<0.05) and small intestine contents in the broilers was significantly increased (P<0.05) as the levels of wormwood addition increased. Therefore, although there was no significant improvement for the performance of broiler chickens with the dietary supplementation of wormwood meal, less than 1% addition of wormwood to broiler diets might have beneficial for human health by reducing the abdominal fat deposition of the broiler chicknens.

• Clinical and Experimental Pediatrics
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• v.46 no.12
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• pp.1212-1216
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• 2003

Purpose : Rotavirus is a leading cause of severe gastroenteritis in infants and young children around the world. The aim of this study is to investigate the fat content in stools of patients with rotaviral enteritis compared to the stools of children who had no gastroenteritis. Methods : Seventy two patients who were admitted to Konkuk University Hospital, College of Medicine from Jun 2001 to May 2002 due to rotaviral enteritis and seventy five patients who were admitted at the same time with other diseases with no gastrointestinal problems as control, were enrolled in this study. The age of patients was from one month to five years. The average age of children with rotaviral enteritis was $17{\pm}11months$ and the average age of control patients was $14{\pm}15months$. Fat content of stools was investigated by acid steatocrit tests in both patients with rotaviral enteritis and control. Results : Acid steatocrit value of patients with rotaviral enteritis was higher than that of control patients. There was no difference in acid steatocrit value of children with rotaviral enteritis among the age groups. In one month- to six month-old infants, there was no difference in acid steatocrit values between the children with rotaviral enteritis and control patients. But, over the age of seven months, the acid steatocrit value of children with rotaviral enteritis was higher than that of control patients. Conclusion : We are of the opinion that fat malabsorption in patients with rotaviral enteritis and steatorrhea in rotaviral enteritis may result from decreased fat absorption in the small intestine.

• Journal of Korean Medical classics
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• v.12 no.1
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• pp.226-254
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• 1999

For the first time, the theory of ChonKwanChuk(寸關尺) and the examination method of KyungJung(輕重法) of KiGu(氣口) were formed in "NanKyung(難經)". After that, the vicera assignment theory at left and right hands(左右守藏府記屬理論) was established in "MaekKyung(脈經)" After Reserching the theoretical relevance of the two books, theories between the doctors who understood the examination method of ChukBu(尺部診法) of "Somun MaekYoJungMiRon(素問 脈要精微論)" as the examination method of KyungJung(輕重法) and the doctors who understood that method as the method of ChonKwanChuk(寸關尺) were compared and researched. The results were as follows : 1. The posit ion of ChonKwanChuk(寸關尺) of the examination method of ChonKwanChuk(寸關尺法) is explained at 2nd Nan(二難) of "NanKyung" as follows. Chuk(尺) is the position which is 1Chon(1寸, unit) distant from Kwan(關) to the direction of ChukTaek acupuncture point(尺澤穴) and Chon(寸) is the position which is 9Pun(9分, unit) distant from Kwan(關) to the direction of EoJe acupuncture point(魚際穴). And the six vessels(六經) were assined to ChonKwanChuk(寸關尺) on the basis of OHangJaMoSnagSaeng(五行子母相生) at 18th Nan(18難) of NanKyung. After that Yang Hyun-Jo(楊玄操) at Dang Dynasty, Jung Deok-Yong(丁德用), Woo Seo(廬庶) at Song dynasty explaind the examination method of ChonKwanChuk(寸關尺法) of NanKyung as the method of ChonKwanChuk of two hands(兩手寸關尺法) from the viewpoint of "MaekKyung(脈經)". 2. From the viewpoint of MaeKyung, the vicera assignment of the two hand ChonKwanChuk method is as follows. At Chuk of left hand, the Heart and Small intestine are assigned. At Kwan of left hand, the Liver and Gall Bladder are assigned. At Chuk of left hand, the Kidney and Bladder are assigned. At Chuk of right hand, the Lung and Large in testine are assigned. At Kwan of right hand, the Spleen and Stomach are assigned. At Chuk of Right hand, the Vital Gate(命門) and Bladder are assigned. 3. For the first time, HwalSu(滑壽) at Won dynasty said that the paragraph "尺內兩傍${\cdots}{\cdots}$" of "Somun MaekYoJungMiRon" is the first of the examination method of KiGu(氣口診脈法). After that Ma Shi(馬蒔), Jang Gae-Bin(張介賓) of Myung Dynasty, Jang Ji-Chong(張志聰), Seo Dae-Chun(徐大椿) of Chung Dynasty who were influenced by him explained that paragraph as the method of KiGuChonKwanChuk(氣口寸關尺法). 4. Lee Kyu-Jun(李圭晙) explained the paragraph "尺內兩傍${\cdots}{\cdots}$" of "Somun MaekYoJungMiRon" as the method of KyungJung(輕重法) and explained Chok(尺) as the paragraph 'Chuk is low position of Kigu, and it means the depth(氣口之下位也, 言其深也)' and explained that 'the Left and the Right is layers(左右者層數). And he revised that the Jang(臟) must be examined at the inner part and the Bu(府) must be examined at the outer part. By this, he settled the theoratical basis of the method of KyungJung(輕重法). 5. The doctors who used the examination method of ChonKwanChok(寸關尺診法) settled their logical justification of the two hand examination method of ChonKwanChuk(兩手寸關尺診法) by connecting with "Somon MaekYoJungMi-Ron" from the viewpoint of 2nd Nan(難) and 18th Nan(難) of NanKyung and MaekKyung. On the contrary, the doctors who used the examination method of KyungJung(輕重診法) settled their logical justification of the examination method of KyungJung(輕重診法) by connecting with "Somun MaekYoJungMiRon" from the viewpoint of 4th Nan(四難) and 5th Nan(五難).

• Advances in pediatric surgery
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• v.12 no.1
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• pp.70-85
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• 2006

A nationwide survey on necrotizing enterocolitis (NEC) was undertaken among members of the Korean Association of Pediatric Surgeons. The members were required to complete a questionnaire and the case registration form for each patient during the three-year period from July 2001 to June 2004. Eighty one patients were included in the questionnaire, but only 71 patients were registered from 22 members in 16 hospitals. At the same time survey on focal intestinal perforation (FIP) was undertaken and 17 patients were registered from 11 members in 10 hospitals. Total 19,041 newborns were admitted to neonatal intensive care unit during the study period. Eighty one patients (17.27 %) were underwent surgery among 469 babies who were managed under the impression of NEC. The male to female ratio was 2.1:1. The premature were 60.6 %. The most common site of involvement was ileum. Overall and operation survival of NEC were 72 % and 79 %, respectively. The survival was lower in smaller babies, multiple segments involvement and involvement of both the small and large intestine. But there was no difference in survival according to sex or prematurity. The FIP showed very similarity with NEC in terms of incidence, and the age of diagnosis and operation. But the survival was much better and 100 %. The results showed the clinical characteristics of NEC and the trend of management in NEC in Korea. In the future we hope we can discuss about this topic in prospective manner.

• Journal of Life Science
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• v.19 no.9
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• pp.1226-1231
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• 2009

Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.

• Journal of Life Science
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• v.28 no.4
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• pp.389-397
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• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.