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Biochemical Characterizations of Phenylalanine Ammonia-Lyase and its Mutants to Develop an Enzymatic Therapy for Phenylketonuria (페닐케톤뇨증의 효소치료 개발을 위한 phenylalanine ammonia-lyase 및 유전자 변이형의 생화학적 특성)

  • Kim, Woo-Mi
    • 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.

Hemodynamic Instability after Patent Ductus Arteriosus Ligation in Very Low Birth Weight Infants (극소저체중출생아에서 동맥관 개존증 결찰술 후 발생한 혈역동학적 불안정성에 대한 고찰)

  • La, Kyong-Suk;Lee, Jang-Hoon;Choi, Byung-Min;Han, Heon-Seok;Hong, Young-Sook;Lee, Joo-Won
    • Neonatal Medicine
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    • v.17 no.2
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    • pp.201-206
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    • 2010
  • Purpose: Recently, after patent ductus arteriosus (PDA) ligation in preterm infants, unexplained hemodynamic instabilities are reported. To determine the incidence, risk factors and clinical manifestations of hemodynamic instability after PDA ligation in very low birth weight (VLBW) infants. Methods: This retrospective multicenter study enrolled 18 VLBW infants who underwent PDA ligation from January 2002 to February 2008. Hemodynamic instability defined as unexplained cardiopulmonary dysfunction with increased dependency on mechanical ventilation and decreased blood pressure. Results: The mean gestational age and birth weight (BW) of all infants were $27^{+6}{\pm}1^{+6}$ weeks and 951${\pm}$245 g. Hemodynamic instability group (HI) included seven infants (39%) and hemodynamic stability group (HS) included 11 infants (61%). Compared to HS, HI had lower BW (1,033${\pm}$285 g vs. 821${\pm}$126 g, P=0.048) and weight on operation day (1,195${\pm}$404 g vs. 893${\pm}$151 g, P=0.042), longer hospital days (105${\pm}$29 vs. 141${\pm}$39, P=0.038), more severe bronchopulmonary dysplasia (BPD), (no/mild/moderate/severe, 2/5/2/2 vs. 0/1/2/4, P=0.038) and higher preoperative $FiO_2$ (0.29${\pm}$0.06 vs. 0.38${\pm}$0.09, P=0.02). One case of mortality due to sepsis, which was not associated with ligation, was observed among HS. Conclusion: The incidence of hemodynamic instability after PDA ligation in VLBW infants was 39%. Low BW, low weight on operation day and preoperative high $FiO_2$ might be risk factors of hemodynamic instability after PDA ligation in VLBW infants. The hemodynamic instability could increase the severity of BPD and hospital days.

Anti-diabetic Effects of Barnyard Millet Miryang 3 [Echinochloa esculenta (A. Braun)] Grains on Blood Glucose in C57BL/KsJ-db/db Mice (식용피 밀양3호[Echinochloa esculenta (A. Braun)] 에탄올 추출물의 당뇨모델 마우스에 대 한 항당뇨 활성)

  • Kwon, Gi Hyun;Jun, Do Youn;Lee, Ji Young;Park, Jueun;Woo, Mi Hee;Yoon, Young Ho;Ko, Jee Youn;Oh, In-Seok;Kim, Young Ho
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1265-1272
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    • 2015
  • Barnyard millet Miryang 3 [Echinochloa esculenta (A. Braun)] grains have recently been acknowledged for beneficial health properties due to phenolic ingredients and dietary fiber. This study has been conducted on the anti-diabetic activity of barnyard millet Miryang 3 which shows the strongest anti-inflammatory activity among barnyard millet inhabiting in South Korea. When 80% ethanol (EtOH) extract of barnyard millet Miryang 3 grains were orally administered into db/db diabetic mice for 8 weeks (600 mg/kg/day), the glucose level in blood following fasting appeared to be improved compared to the control group. The results of glucose tolerance test and blood lipid profile assay were similar to those of the metformin-administered positive control group. In addition, the level of body weight increase (8.54±2.24) was lower than the level of metformin-administered group (10.36±3.15); however, there was no subtle difference with negative and positive control groups in terms of food efficiency rates. In addition, total cholesterol levels of the 80% EtOH extract-administered group (160.7±7.6) were significantly reduced compared to the diabetic control group (229.3±47.8) and metformin-administered group (176.0±25.6). Consequently, these results show that barnyard millet grains alleviates many of the diabetic symptoms in vivo non-insulin-dependent diabetes mellitus, and suggest that barnyard millet grains can be applicable in developing new functional food materials.

THE CLASSIFICATION OF ADOLESCENTS IN RUNAWAY SHELTERS BY THE EVALUATION OF THEIR PSYCHOPATHOLOGY (보호시설 가출청소년의 정신병리에 대한 평가와 분류)

  • Lee, Jong-Sung;Kwack, Young-Sook
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.12 no.2
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    • pp.192-217
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    • 2001
  • Object:This study was carried out to classify adolescents in runaway shelters by evaluating their psychopathology. And the ultimate purpose is to offer basic data for preventing adolescents‘ runaway and for diversifying runaway shelters suitable for the problem of individual adolescent. Method:128 adolescents who stay in the runaway shelters were asked to complete self-report qeustionnaires including basic sociodemographic data, Child Behavior Check List(CBCL), Minnesota Multiphasic Personality Inventory(MMPI), and Symptom Check List-90-Revised(SCL-90-R). Korean Wechsler Adult Intelligence Scale(K-WAIS)[or Korean Educational Developmental Institute-Wechsler Intelligence Scale for Children(KEDI-WISC)] and Bender-Gestalt test(BGT) were also done by clinical psychologists. Results:The most common age of the subjects were 15-year-old, and they dropped out their schools in the middle school most commonly. Mostly they were from middle class family and their parents' educational level were high school graduates. The first runaway episode was most common in the middleschool period, and their runaways were repeated. The most common frequency of runaways were more than 10 times. About 10% of them abused drugs and about 80% of them abused alcohol. One third of them had experiences of illegal problems and 10% of them engaged in sexual activity for money. 95 adolescents(83%) in CBCL, 42 adolescents(36%) in SCL-90-R, and 70 adolescents(69.3%) in MMPI showed clinical significance. In intelligence test, 22 adolescents(22%) were mentally retarded. In BGT, 35 adolescents(39.4%) manifested brain dysfunction signs. Conclusion:Runaway adolescents in the shelters have variable and severe psychopathology. Their psychopathology is classified as follows;The behavior disorder group, the mood disorder group with anxiety/depression, the somatic disorder group with somatic symptoms, and the psychosis group with possibility of severe psychopathology. Therefore it is very important to evaluate psychiatric problems of runaway adolescents, and specific therapeutic interventions according to their problems are required.

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The Cytological Study of the Experimental Middle Ear Effusion. (실험적 중이삼출액의 세포학적 고찰)

  • 심상열;문태용;윤강묵;박순일;박인용
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1982.05a
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    • pp.13.2-14
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    • 1982
  • Otitis media with effusion, described first by Politzer (1867), is closely related with the function of auditory tube, but its etiology and pathogenesis are not clearly defined yet. There are many theories about its pathogenesis including hydrops ex vacuo theory which was most reliable nowadays. In this paper, using cats in experimental animals, hydrops ex vacuo theory was proved and cytological study of the effusion and light microscopic observation of the middle ear mucosa in otitis media with effusion were done. The results were as follows: 1) The effusion was found in all experimental groups after eighteen hours of the auditory tube obstruction. 2) In the cytological study of effusion by smear technic, Polymorpholeukocytes were dominant in earlier days but monoculear cells were soon increased and no eosinophils were found. 3) In the culture of the effusion, no bacteria was cultured. 4) By opeating microscope, hypertrophy of the middle ear mucosa observed especially in the fourteen days after auditory tube obstruction and effusion was most remarkable in the fourteen days, also. 5) By light microscopy, there were epithelial hyperplasia, proliferation of goblet cells, capillaries and infiltration of inflammatory cells which showed same distribution as smear technic.

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A Dynamic Behavior Evaluation of the Curved Rail according to Lateral Spring Stiffness of Track System (궤도시스템의 횡탄성에 따른 곡선부 레일의 동적거동평가)

  • Kim, Bag-Jin;Choi, Jung-Youl;Chun, Dae-Sung;Eom, Mac;Kang, Yun-Suk;Park, Yong-Gul
    • Proceedings of the KSR Conference
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    • 2007.11a
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    • pp.517-528
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    • 2007
  • Domestic or international existing researches regarding rail damage factors are focused on laying, vehicle conditions, driving speed and driving habits and overlook characteristics of track structure (elasticity, maintenance etc). Also in ballast track, as there is no special lateral spring stiffness of track also called as ballast lateral resistance in concrete track, generally, existing study shows concrete track has 2 time shorter life cycle for rail replacement than ballast track due to abrasion. As a result of domestic concrete track design and operation performance review, concrete track elasticity is lower than track elasticity of ballast track resulting higher damage on rail and tracks. Generally, concrete track has advantage in track elasticity adjustment than ballast track and in case of Europe, in concrete track design, it is recommended to have same or higher performance range of vertical elastic stiffness of ballast track but domestically or internationally review on lateral spring stiffness of track is very minimal. Therefore, through analysis of service line track on site measurement and analysis on performance of maintenance, in this research, dynamic characteristic behaviors of commonly used ballast and concrete track are studied to infer elasticity of service line track and experimentally prove effects of track lateral spring stiffness that influence curved rail damage as well as correlation between track elasticity by track system and rail damage to propose importance of appropriate elastic stiffness level for concrete and ballast track.

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Anti-inflammatory Activity of Crinum asiaticum Linne var. Japonicum Extract and its Application as a Cosmeceutical Ingredient (문주란의 항염효과와 화장료적 특성)

  • Kim, Ki-Ho;Kim, Young-Heui;Kim, Ki-Soo;Park, Sun-Hee;Lee, Soo-Hee;Kim, Young-Jin;Kim, Young-Sil;Kim, Jong-Heon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.59-64
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    • 2006
  • Crinum asiaticum Linne var. japonicum has long been used as a rheumatic remedy, an anti-pyretic, an anti-ulcer treatment, and for the alleviation of local pain and fever in Korea and Malaysia. In order to investigate the possibility of Crinum asiaticum Linne var. japonicum extract as a cosmetic ingredient, we measured its anti-inflammatory effect by inhibition of iNOS (inducible nitric oxide synthase), and the release of PGE2, IL-6, and IL-8. HPLC experiment after extraction with 95% ethanol at pH 3.5 showed that Crinum asiaticum Linne var. japonicum was mainly composed of lycorine (up to 1%), a well-known immunosuppressant. The content of lycorine varied depending on the type of tissue analyzed and the extraction method. In anti-inflammatory assay for inhibition of nitric oxide formation on lipopolysaccharide (LPS)- activated mouse macrophage RAW 264.7 cells, the ethanolic extract of Crinum asiaticum showed inhibitory activity of NO production in dose-dependent manner ($IC_{50} = 83.5 {\mu}g/mL$). Additional study by RT-PCR demonstrated that the extract of Crinum asiaticum significantly suppressed the expression of the iNOS gene. Moreover, the extract of Crinum asiaticum did not show my cytotoxicity, but did show cell proliferation effect against LPS ($10{\sim}60%$ increase of tell viability). In an assay to determine inhibition of the $H_2O_2$-activated release of PGE2, IL-6, and IL-8 in human normal fibroblast cell lines, the release of PGE2 and IL-6 was almost completely inhibited above concentrations of 0.05% and 1%, respectively. Moreover, the release of IL-8 was completely inhibited over the entire range of concentrations (> 0.0025%). The result showed that the extract of Crinum asiaticum Linne var. japonicum has sufficient anti-inflammatory effect. There-fore, Crinum asiaticum Linne var. japonicum extract may be useful as an ingredient of cosmetic products.

Factors for the Prediction of Pain in Terminally Ill Cancer Patients in Hospice Units (호스피스 병동에 입원한 말기 암환자의 통증 예측요인)

  • Yong, Jin-Sun;Han, Sung-Suk;Ro, You-Ja;Hong, Hyun-Ja
    • Journal of Hospice and Palliative Care
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    • v.5 no.2
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    • pp.125-135
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    • 2002
  • Purpose : The purpose of this study was to investigate the impact of depression, discomfort, spirituality, physical care, and opioid use on pain with terminally ill cancer patients in the hospice units. Method : The convenient sample of this study consisted of 58 terminally ill cancer patients at three hospice units in university-affiliated hospitals. Patients were interviewed with structured questionnaires. The data was analyzed using ANOVA, Pearson correlation coefficient, and multivariate multiple regression. Result : The results of this study were as follows : 1) The mean age of the participants was approximately 57 years. Regarding diagnosis, stomach cancer showed the highest frequency (24.1%), followed by lung cancer (17.2%) and rectal cancer (13.8%). Regarding motivation for admission to the hospice unit, the majority of the participants indicated pain control (67.2%), followed by spiritual care (39.7%), and symptom relief (27.6%). 2) The mean pain level measured by VAS was 5.13 (${\pm}2.61$). Regarding pain type, the highest pain frequency the participants experienced was deep pain (53.4%), followed by multiple pain (20.7%), intestinal pain (17.3%), and neurogenic (5.2%) and superficial pain (3.4%). 3) Regarding the factors influencing pain, the pain level was significantly affected by the depression level (P<0.01) and the opioid use (P<0.01). Conclusion, In summary, the higher the level of pain the terminally ill cancer patents had the higher the depression level as well as the opioid use. Thus, health care professionals need to continuously provide holistic care for them to die comfortably.

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A Clinical Study on Factors affecting Spontaneous Resolution of Vesicoureteral ,Reflux in Children (방광요관역류의 완해에 영향을 미치는 인자에 관한 연구)

  • Song, Byung-Ho;Oh, Chang-Weon;Kim, Ki-Bok
    • Childhood Kidney Diseases
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    • v.2 no.2
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    • pp.152-160
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    • 1998
  • Purpose : Vesicoureteral reflux (VUR), which can result in renal failure in children, is expected to resolve spontaneously in mild cases, but severe cases require surgical correction, posing dilemma in deciding the right measures in some cases. Therefore, to delineate the factors affecting spontaneous resolution of VUR and to apply them in assessing the prognosis, we reviewed the patients with VUR who had been treated medically. Patients and Methods : 32 children (49 renal units) with reflux who had been admitted during the 5-year period from Jan '92 to Dec '96 were classified into 4 groups (Resolved, Improved, Unchanged, and Worsened) and analyzed. Results : 1) 25 were boys and 7 girls. The age at diagnosis ranged from 13 days to 9 years (mean $24.6{\pm}11.4$ mo) 2) Among 49 refluxing ureters, 4 were below 1 me of age,21 between 1 me to 1 yr, 16 between 1 yr to 6 yr, 8 beyond 6 yr. Two belonged to Grade I, 16 to Crade II, 17 to Grade III, 12 to Grade IV, and 2 to Grade V. 3) Spontaneous resolution rates of reflux were 100, 81, 47, 8, 0$\%$ for each Grade, respectively. Resolution plus improvement rates in Grade III and IV were 71 and $50\%$, each. 4) Resolution rates in relation to the age at diagnosis were $100\%$ for below 1 mo, $48\%$ between 1 mo to 1 yr, $56\%$ between 1 to 6 yr, and $13\%$ beyond 6 yr. 5) Of 15 unilateral refluxing ureters, 14 ($93\%$) resulted in resolution. Of 34 bilateral refluxing ureters, 11 ($32\%$) resolved spontaneously 6) Resolution occurred within 1 year in 20 units of 24 renal units in regular follow-up. Conclusion: The lower the initial grade and the younger the patient, the sooner the reflux resolved. Resolution was better when VUR was unilateral than bilateral. Long-term regular follow-up is essential because even the cases falling in Grades III and IV have high rates of spontaneous resolution and improvement with medical treatment.

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Effect of Unilateral Renal Perfusion of Cyclosporine and Mitomycin on Rat's Kidney (Cyclosporine과 Mitomycin의 일측성 신관류로 초래되는 백서 신병변에 관한 연구)

  • Baek Seung In;Lim Hyun Suk;Shin Weon Hye;Ko Cheol Woo;Koo Ja Hoon;Kwak Jung Sik
    • Childhood Kidney Diseases
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    • v.2 no.2
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    • pp.138-144
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    • 1998
  • Purpose : The use of cyclosporine and mitomycin in various immunologic or neoplastic disorders has been known to cause wide-ranged nephrotoxic effects including thrombotic microangiopathy. However, the mechanism of nephrotoxicity of these drugs has not been studied adequately, so that present experimental study has been undertaken to find out whether these drugs can cause direct damage to the kidney and to clarify the pathogenetic mechanism of nephrotoxic effect of these drugs. Materials and methods : Sprague-Dawley rats weighing 250-300 gm were used for experimental animals and unilateral renal perfusion technique, modified from the method described by Hoyer et al was used. Isolation of left kidney from systemic circulation was made by clamping aorta and left renal vein and a hole was punctured in the anterior wall of the left renal vein. Cyclosporine (2.5 mg in 4 ml solution) and mitomycin (1.6 mg in 4ml solution) were infused through left renal artery and normal saline was used in control rats. Forty-eight hours after infusion of the drugs, animals were sacrificed and left kidney removed and processed for histologic examination. Total ischemic time of left kidney was less than 15 minutes: Results : Cyclosporine-perfused group showed severe swelling of glomerular endothelial ceil along with swelling of glomerular epithelial cell and interstitial vascular endothelial cell. Mitomycin-perfused group also showed severe swelling of glomerular endothelial and epithelial cells. And in addition to these findings, they demonstrated platelets aggregation, swelling and degranulation of platelets and fibrin accumulation in some of the capillaries, indicating occurrance of thrombotic microangiopathy. Conclusion : present experiment indicates that cyclosporine and mitomycin can cause direct toxic injury to renal endothelial cell. And this direct toxic damage to endothelial cell seems to be an important initiating event for the development of thrombotic microangiopathy.

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