• Title/Summary/Keyword: membrane analysis

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Antimicrobial, Antioxidant and Cellular Protective Effects of Houttuynia cordata Extract and Fraction (어성초 추출물 및 분획물의 항균, 항산화 및 세포보호활성)

  • Yun, Mid Eum;Lee, Ye Seul;Lee, Yun Ju;Park, Young Min;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.4
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    • pp.452-460
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    • 2018
  • This study was conducted to investigate the physiological activities of Houttuynia cordata extracts and fractions. H. cordata extracts were extracted with 50% ethanol and the ethyl acetate fractions were obtained from the extracts. Minimum inhibitory concentration (MIC) values of the ethyl acetate fraction for S. aureus and B. subtilis were $78{\mu}g/mL$ and $312{\mu}g/mL$, respectively, indicating the high activity against gram-positive bacteria. The free radical scavenging activity ($FSC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl (DPPH) was higher in the ethyl acetate fraction with $12.00{\mu}g/mL$ compared to that of $27.15{\mu}g/mL$ for 50% ethanol extract. The total antioxidant activity ($OSC_{50}$) values for reactive oxygen species (ROS) produced in $Fe^{3+}-EDTA/H_2O_2$ system by a luminol-dependent chemiluminescence method were 2.91 and $0.983{\mu}g/ml$ for the 50% ethanol extract and ethyl acetate fraction, respectively. To investigate cellular protective effects on the HaCaT cell, the intracellular ROS scavenging activity was measured after UVB irradiation and the ethyl acetate fraction of H. cordata showed the activity in a concentration-dependent from $1.6{\mu}g/mL$ and a reduction rate of 54.3% at a maximum concentration of $12.5{\mu}g/mL$. Also, HaCaT cell protective effect against $H_2O_2$-mediated decreased the cell viability of the ethyl acetate fraction of H. cordata which significantly increased the cell viability from $0.8{\mu}g/mL$ and the maximum cell viability showed 86.9%. The ethyl acetate fraction of the H. cordata extracts was analyzed by TLC and HPLC. As a result, quercitrin, isoquercitrin, hyperoside, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, rutin and afzelin were identified. From the above results, it was suggested that the extracts and fractions of H. cordata have a potential to be applied in the field of cosmetics as a natural antioxidant/preservative capable of protecting the cell membrane from the oxidative stress by eliminating ROS and exhibiting the antimicrobial effect.

Antioxidative, and Inhibitory Activities on Melanogenesis of Vitex negundo L. Leaf Extract (목형 잎 추출물의 항산화 활성과 멜라닌 생합성에 대한 저해활성)

  • Kim, A Reum;Park, Su Ah;Ha, Ji Hoon;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.135-144
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    • 2013
  • The aim of this study was to evaluate various aspects of Vitex negundo L. leaf extract, such as the antioxidative activity, tyrosinase inhibitory effects, and inhibitory activities on ${\alpha}$-MSH induced melanogenesis, and active component analysis. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) scavenging activities ($FSC_{50}$) of the ethyl acetate fraction and aglycone fraction of V. negundo L. leaf extract were $14.51{\mu}g/ml$ and $13.96{\mu}g/ml$, respectively. A luminol-dependent chemiluminescence assay revealed that the reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of the aglycone fraction of V. negundo L. leaf extract on ROS generated in an $Fe^{3+}$-$EDTA/H_2O_2$ system was the most prominent at $0.22{\mu}g/ml$. The protective effects of the extracts fractions of V. negundo L. leaf against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim}50{\mu}g/ml$). In particular, there were greater protective effects of the aglycone fraction on the cellular membrane than that of the fat-soluble antioxidant (+)-${\alpha}$-tocopherol. The inhibitory effects ($IC_{50}$) on mushroom tyrosinase were the highest for the ethyl acetate fraction ($IC_{50}$ = $48.58{\mu}g/ml$). The inhibitory effect on ${\alpha}$-MSH induced melanogenesis in B16 melanoma cells was 41.80% at $50{\mu}g/ml$ of ethyl acetate fraction. Active component analyses by TLC, HPLC and LC/ESI-MS revealed luteolin and isoorientin. These results indicate that V. negundo L. leaf extract can be used as an antioxidant for ROS scavenging. Particularly, the luteolin and isoorientin of the ethyl acetate fraction may be applicable to new whitening cosmetics because of its inhibitory effect on mushroom tyrosinase and ${\alpha}$-MSH induced melanogenesis in B16 melanoma cells.

Gastric Dilatation and Volvulus in Shih-tzu (시츄에서 발견된 위확대염전)

  • Moon, Joon Ho;Kim, Hyun Ah;Ryu, Minok;Jang, Min;Ji, Seoyeon;Lee, Inhyung;Yoon, Junghee;Choi, Mincheol;Youn, Hwayoung;Lee, Byeong Chun;Jang, Goo
    • Journal of Veterinary Clinics
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    • v.32 no.3
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    • pp.255-258
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    • 2015
  • Gastric dilatation and volvulus (GDV) is an acute and life-threatening disease most commonly affecting large- and giant-breed dogs. However a 17-year-old Shih-tzu (4 kg, spayed female) was hospitalized for acute GDV. Repeated unproductive retching, lethargy, and excessively enlarged abdomen were observed. Physical examination indicated that the patient had suffered from hypothermia ($36.5^{\circ}C$), tachycardia (240 bpm), slowed capillary refill time (> 2 sec.), and pale mucous membrane. Grade III murmur with normal lung sound was auscultated. Abdominal palpation revealed that tympanic regions existed in both the left and right sides. Systolic blood pressure decreased gradually from 220 to 40 mmHg within 4 hours. In blood analysis, slight azotemia was observed by blood urea nitrogen (BUN; 29.1 mg/dl) and creatinine (1.6 mg/dl). Blood lactate concentration (8.13 mmol/l) was severely elevated. Additionally, dilatation and volvulus of the stomach was observed by radiograph. Supportive oxygen, heat, fluid, and drugs were administered with gastric decompressions (e.g., gastrocentesis and nasogastric tube). However the patient entered into comatose status with uncontrollable systolic blood pressure, despite the administration of dobutamine intravenously. The case was closed by euthanasia, considering welfare and age. We finally diagnosed the patient as a GDV, thus this is the first GDV case report in small-breed dog such as Shih-tzu.

Clinical Characteristics of Bronchopulmonary Dysplasia by Type and Severity (기관지폐이형성증의 유형 및 중증도에 따른 임상적 특징)

  • Shim, Gyu-Hong;Lee, Hyun-Ju;Kim, Eun-Sun;Lee, Jin-A;Choi, Chang-Won;Kim, Ee-Kyung;Kim, Han-Suk;Kim, Beyong-Il;Choi, Jung-Hwan
    • Neonatal Medicine
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    • v.17 no.1
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    • pp.21-33
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    • 2010
  • Purpose : The increased survival of preterm infants in the neonatal intensive care unit recently has resulted in an increased frequency of bronchopulmonary dysplasia (BPD), especially with atypical forms. However, there have been few studies compairing classic and atypical BPD. The aim of this study was to investigate the differences between these two types of BPD. Methods : Infants with a gestational age less than 32 weeks born at the Seoul National University Hospital and Bundang Seoul National University Hospital from May 2004 to April 2007 were included. The data were categorized in 2 groups, classic and atypical BPD. We determined the incidence of BPD, and compared perinatal factors and postnatal managements between two groups. Results : Among 260 study subjects, 141 (54.2%) infants had BPD. Classic BPD infants were 64 and atypical BPD infants were 77. Comparison of differences between 2 groups, classic BPD infants were associated with respiratory distress syndrome, patent ductus arteriosus, intrauterine growth restriction, more high-frequency ventilator (HFV) use, low 1 and 5 minute Apgar scores. Atypical BPD infants were associated with antenatal steroid use, maternal premature rupture of membrane and chorioamnionitis (CAM). In multivariate analysis, more HFV use was associated with classic BPD. Antenatal steroid use, clinical CAM and histological CAM were associated with atypical BPD. Conclusion : The results of this study showed that antenatal factors (antenatal steroid use, clinical CAM, histological CAM) were associated with atypical BPD and postnatal factors (HFV used more) were associated with classic BPD. Further studies are needed for prevention and treatment of BPD.

Effects of Characteristics of Ovarian follicular Fluid and Ant-Inhibin Serum on Steroid Hormone Secretion by Hanwoo Granulosa Cells In Vitro (한우 난소의 Follicular Fluid의 특징과 과립막 세포의 스테로이드호르몬 분비에 대한 Anti-Inhibin Serum의 첨가효과)

  • 성환후;민관식;양병철;노환국;최선호;임기순;장유민;박성재;장원경
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.119-124
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    • 2001
  • This study was performed to investigate the effects of the peptide to carrier ratio on the immune and biological functions to inhibin immunization in Hanwoo. A peptide sequence kom the alpha -subunit (19~32 peptide) of porcine inhibin was synthesized for antigen and conjugated to human serum albumin(HSA) for carrier protein. Anti-inhibin sera(AI) were produced 52 day later from rabbit after injection of inhibin-$\alpha$ -subunit peptide conjugator for antigen with the interval of 2 weeks. Immune-blotting analysis using antibody specific fur inhibin-$\alpha$ subunits revealed that the inhibin was detected at 1.0 cm bovine follicular fluid(bFF). However, each stage of corpus lutea and 0.1 cm of follicular fluid were not detected. The maximal contents of estradiol-17 $\beta$ in Hanwoo ovarian follicular fluid were detected at 2.0 cm of follicular size(diameter), but the mean total contents of these hormone decreased significantly with decreasing diameter of follicles. However, progesterone contents of follicular fluid were high at 1.0 cm of follicle. Progesterone secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) inhibited in 5% bFF and 5% bFF + 5% AI addition group compared with control group. Estradiol-17 $\beta$ secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) increased in 5% AI and 5% AI + 5% bFF addtion group compared with control group. However, the groups added 5% AI were not changed compared to control groups in progesterone and estradiol-17 $\beta$. Taken together, we suggested that inhibin in the mature FF plays a pivotal role on the biosynthesis of steroid hormone of follicular cells during follicular development.

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Physiological Characteristics Related to Cold Injury in Rice (수도 냉해에 관련된 생리적 특성 고찰)

  • 석순종;허일봉;임정남
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.36 no.5
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    • pp.429-444
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    • 1991
  • Cold stress influence plant growth through a wide range of growth characters. Adverse effects of low temperature to plant growth come from results of colligative and complex physiological responses to cold stress. To evaluate more exactly cold tolerance of crop plant, it is needed to observe physiological changes induced by cold stress and to analyze relationships between intraspecific variations in physiological factors related to cold tolerance and the extent of cold tolerance in the field. Therefore, the composition and unsaturation ratio of fatty acids in phospholipid, a constituent of membrane, the transition-temperature in respiratory activity of mitochodria, the chlorophyll fluorescence as a factor related to photosynthesis were investigated in rice plant and data on these factors were compared with the degree of cold tolerance obtained in the field experiment. Also, effects of hardening and Mn++ treatment were evaluated as a method to reduce chilling injuries. The unsaturation ratio of fatty acids, whether rice plants were grown in a natural condition or under the chilling stress, was higher in the cold- tolerant varieties and was significantly correlated with the degree of cold tolerance (1-9) observed in the field experiment. And it was also increased by chilling treatment or hardening treatment, due to a reduction in palmitic acid content and an increase in linolenic acid content. The transition-temperature of respiratory activity of mitochodria isolated from etiolated rice seedlings ($25^{\circ}C$, two week-grown in the dark), was correlated with the degree of cold tolerance in the field, cold -tolerant varieties showing a lower transition-temperature. It was not influenced by growth stages. The intensity of chlorophyll fluorescence was highly correlated with the degree of cold tolerance, cold-tolerant varieties having a higher fluorescence intensity. By foliar application of Mn, the transition-temperature of respiratory activity was lowered as much as 0-2$^{\circ}C$ in all tested varieties. Soil application of Mn induced more significant effect in cold-susceptible varieties with a possibility of reducing chilling injuries. On the whole, there were high correlationships among the degree of cold tolerance, the unsaturation ratio of fatty acids in phospholipid, the transition- temperature of respiratory activity and chlorophyll fluorescence except for a few varieties. The transition- temperature of respiratory activity appeared to be negatively correlated with the unsaturation ratio of fatty acids. and the chlorophyll fluorescence to be positively correlated with the unsaturation ratio. This implies that these physical and physiological factors were very closely related to cold tolerance and can be used as an effective index of the evaluation of cold tolerance of crop plant. But other factors as well as three factors discussed above are needed to be considered colligatively and altogether with a systematic analysis for the more exact evaluation of cold tolerance. in rice cultivars. in rice cultivars.

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Studies on the Function of the Rv2435c Gene of the Mycobacterium bovis BCG (Mycobacterium bovis BCG Rv2435c 유전자의 기능에 대한 연구)

  • Lee Seung-Sil;Bae Young-Min
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.415-422
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    • 2005
  • library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

Iron Status According to Serum Selenium Concentration and Physique in Young Female Adults (젊은 여성의 혈청 셀레늄 농도 및 체격에 따른 체내 철 수준)

  • Lee, Ok-Hee;Chung, Yong-Sam;Moon, Jong-Wha
    • Journal of Nutrition and Health
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    • v.43 no.2
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    • pp.114-122
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    • 2010
  • Se and Fe are trace minerals acting as antioxidant scavenging free radicals. Iron deficiency is the most frequently reported nutritional deficiency in females. Body iron status are known to be dependent not only upon dietary iron intake, but also upon micro-mineral nutrition and obesity. Antioxidants such as selenium are reported to play an important role on the regulation of erythropoiesis by protecting RBC membrane from antioxidative damage. In this study, iron status in young females and its relationships with selenium status and physique were examined. Serum selenium and iron concentrations were measured by HANARO research reactor using neutron activation analysis method (NAA-method). The proportion with iron deficiency and anemia were 27.1% and 8.6%, respectively in young females, but the proportion with iron deficient anemia was 1.4%. The mean serum selenium level was $12.0\;{\mu}g/dL$ and in normal range in the young women. The study participants were tertiled according to BMI and serum selenium levels. Serum ferritin and iron levels inclined with increasing BMI tertiles. Serum iron and RBC count were higher in middle selenium group than low selenium group. Individuals had significantly lower hematocrit level in the lowest tertile for their serum selenium levels compared with the highest tertile. The serum ferritin level was predicted 25% by BMI and RBC count 26.2% by the serum selenium level and body fat%. In conclusion, this study shows that body iron status in young adult females are influenced by obesity and body selenium status.

Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocytes for Fat-Reduced Hanwoo Beef Production (한우 체지방 감소 쇠고기 생산을 위한 복강 및 피하지방 항체 개발)

  • Choi, Chang-Weon;Kim, Yu-Hyun;Kim, Sang-Jin;Song, Man-Kang;Kwon, Eung-Gi;Oh, Young-Kyoon;Hong, Seong-Koo;Choi, Seong-Ho;Baek, Kyung-Hoon
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.651-659
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    • 2008
  • This study aimed to develop polyclonal antibodies to regional inedible adipocytes of Korean native cattle (Hanwoo) and investigate cross-reactivity of the antibodies. Patterns in plasma membrane proteins (PMPs) from abdominal and subcutaneous adipocytes of Hanwoo isolated by collagenase digestion were investigated using SDS-PAGE. As antigens, abdominal and subcutaneous adipocyte PMPs of Hanwoo were injected to sheep 3 times at 3 wk intervals for passive immunization, and non-immunized serum and antisera were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with heart, kidney, liver, lung, muscle, and spleen of Hanwoo were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes of Hanwoo were performed for analysing LDH concentration. Based on the SDS-PAGE analysis, specific proteins of PMPs in abdominal and subcutaneous adipocytes appeared despite rather similar patterns between both adipocytes. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas the antisera had relatively strong reactivity up to the level of 1:128,000 and 1:64,000 dilution. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivities of abdominal and subcutaneous adipocyte antisera were detected with PMPs of the organs. Both antisera strongly reacted with each adipocyte PMPs and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and have safety in cross-reactivities with body organs. Further studies on in vivo cross-reactivity and fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo should be required for inedible fat-reduced high quality beef production.

The Effect of Surfactant Therapy for Acute Lung Injury Induced by Intratracheal Endotoxin Instillation in Rats (기관내 내독소 투여로 유발된 흰쥐의 급성폐손상에서 surfactant의 치료효과)

  • Kang, Yun-Jung;Park, Yong-Bum;Jee, Hyun-Suk;Choi, Jae-Chol;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung-Whui
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.4
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    • pp.487-499
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    • 2000
  • Background : Acute lung injury is an hypoxic respiratory failure resulting from damage to the alveolar-capillary membrane, which can be developed by a variety of systemic inflammatory diseases. In this study the therapeutic effects of intra-tracheal pulmonary surfactant instillation was evaluated in the intratracheal endotoxin induced acute lung injury model of a rat. Methods : Twenty Sprague-Dawley rats were divided into 4 groups, and normal saline (2 ml/kg, for group 1) or LPS (5 mg/kg, for group 2, 3, and 4) was instilled into the trachea respectively. Either normal saline (2 ml/kg, for group 1 & 2, 30 min later) or bovine surfactant (15 mg/kg, 30 min later for group 3, 5 hr later for group 5) was instilled into the trachea. The therapeutic effect of intratracheal surfactant therapy was evaluated with one chamber body plethysmography (respiratory frequency, tidal volume and enhanced pause), ABGA, BAL fluid analysis (cell count with differential, protein concentration) and pathologic examination of the lung. Results : Intratracheal endotoxin instillation increased the respiration rate decreased the tidal volume and int creased the Penh in all group of rats. Intratracheal instillation of surfactant decreased Penh, increased arterial oxygen tension, decreased protein concentration of BAL fluid and decreased lung inflammation at both times of administration (30 minute and 5 hour after endotoxin instillation). Conclusion : Intratracheal instillation of surfactant can be a beneficial therapeutic modality as discovered in the acute lung injury model of rats induced by intratracheal LPS intillation. It deserves to be evaluated for treatment of human acute lung injury.

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