• Title/Summary/Keyword: SacII

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Improvement of Transformation Efficiency Through In Vitro Methylation and SacII Site Mutation of Plasmid Vector in Bifidobacterium longum MG1

  • Kim, Jin-Yong;Wang, Yan;Park, Myeong-Soo;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.1022-1026
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    • 2010
  • The different cleavage patterns of pYBamy59 plasmid isolated from E. coli $DH5{\alpha}$ and B. longum MG1 by the cell extract of B. longum MG1 suggested that the main reason for its low transformation efficiency was related to the restriction modification (R-M) system. To confirm the correlation between the R-M system and transformation efficiency, in vitro methylation and site-directed mutagenesis were performed in pYBamy59. Sequence analysis of pYBamy59 fragments digested by the cell extract of B. longum MG1 revealed that all fragments were generated by restriction of the sequence recognized by SacII endonuclease. When pYBamy59 from E. coli was methylated in vitro by CpG or GpC methyltransferase, it was protected from SacII digestion. Site-directed mutagenesis, which removed SacII sites from pYBamy59, or in vitro methylation of pYBamy59 showed 8- to 15-fold increases in the transformation efficiency over intact pYBamy59. Modification of the SacII-related R-M system in B. longum MG1 and in vitro methylation in pYBamy 59 can improve the transformation efficiency in this strain. The results showed that the R-M system is a factor to limit introduction of exogenous DNA, and in vitro modification is a convenient method to overcome the barrier of the R-M system for transformation.

Ultrastructure of the Ampullate Gland in the Orb Web Spider, Nephila clavata L. Koch II. Sac and Tail Portion of the Large Ampullate Gland (무당거미(Nephila clavata L. Koch) 병상선(甁狀腺)의 미세구조(微細構造) II. 대병상선(大甁狀腺)의 분비낭(分泌囊)과 말단분비부(末端分泌部))

  • Moon, Myung-Jin;Kim, Chang-Shik;Kim, Woo-Kap
    • Applied Microscopy
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    • v.18 no.2
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    • pp.91-101
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    • 1988
  • The ultrastructure of the sac and tail portion of the large ampullate gland and production of the silk materials in the orb web spider, Nephila clavata L. Koch, are studied with electron microscope. Large ampullate glands, the largest glands among the seven kinds of silk glands in this species, are composed of three parts which are the excretory duct, the storage sac and the convoluted tail. The wall of the sac is composed of a single layer of columnar epithelial cells. In the cytoplasm of these cells several kinds of the secretory granules, which are commonly associated with the rough endoplasmic reticula and had characteristic crystalloid, are seen. According to the morphology and internal textures of these granules, the sac portion is subdivided into proximal(connected with the convoluted tail) and distal(connected with excretory duct) portion. Between these two portions, the proximal portion is longer than the distal by four times. Silk materials, being synthesized to the shape of secretory granules, within the glandular epithelial cell of the tail portion are released to the inner cavity by the mechanism of the eccrine secretion. These secretory granules are originated from the rough endoplasmic reticula of the glandular epithelial cells, whereas no Golgi complexes has been found in any of the cells which have been examined.

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Biosynthesis of 3-Hydroxy-5-Methyl-O-Methyltyrosine in the Saframycin/Safracin Biosynthetic Pathway

  • Fu, Cheng-Yu;Tang, Man-Cheng;Peng, Chao;Li, Lei;He, Yan-Ling;Liu, Wen;Tang, Gong-Li
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.439-446
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    • 2009
  • The biosynthesis study of antibiotics saframycin (SFM) in Streptomyces lavendulae and safracin (SAC) in Pseudomonas fluorescens demonstrated that 3-hydroxy-S-methyl-O-methyltyrosine (3hSmOmTyr), a nonproteinogenic amino acid, is the precursor of the tetrahydroisoquinoline molecular core. In the biosynthetic gene cluster of SAC/SFM, sacD/sfmD encodes a protein with high homology to each other but no sequence similarity to other known enzymes; sacF/sfmM2 and sacG/sfmM3 encode methyltransferases for C-methylation and O-methylation; and sacE/sfinF encodes a small protein with significant sequence similarity to the MbtH-like proteins, which are frequently found in the biosynthetic pathways of non ribosomal peptide antibiotics and siderophores. To address their function, the biosynthetic cassette of 3h5mOmTyr was heterologously expressed in S. coelicolor and P. putida, and an in-frame deletion and complementation in trans were carried out. The results revealed that (i) SfmD catalyzes the hydroxylation of aromatic rings; (ii) sacD/sacF/sacG in the SAC gene cluster and sfmD/sfmM2/sfmM3 in the SFM cluster are sufficient for the biosynthesis of 3h5mOmTyr; and (iii) the mbtH-like gene is not required for the biosynthesis of the 3h5mOmTyr precursor.

Effects of Volatile Fatty Acids on IGF-I, IGFBP-3, GH, Insulin and Glucagon in Plasma, and IGF-I and IGFBP-3 in Different Tissues of Growing Sheep Nourished by Total Intragastric Infusions

  • Zhao, Guang-Yong;Sun, Ya-Bo
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.3
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    • pp.366-371
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    • 2010
  • Twelve Suffolk${\times}$Small-tail-Han male sheep (body weight 21-26 kg), aged four months, were used to study the effects of volatile fatty acids (VFA) on IGF-I (insulin-like growth factor-I), IGFBP-3 (insulin-like growth factor binding protein-3), GH (growth hormone), insulin and glucagon in plasma, and IGF-I and IGFBP-3 in different tissues. The sheep were randomly divided into four groups with 3 sheep in each group. The sheep were sustained by total intragastric infusions and four levels of mixed VFA (the molar proportion of acetic acid, propionic acid and butyric acid was 65:25:10), which supplied 333, 378, 423 and 468 KJ energy/kg $W^{0.75}$/d, were infused into the rumen as experimental Treatments I, II, III and IV, respectively. The experiment lasted 12 days, of which the first 8 days were for pretreatment and the last 4 days for collection of samples. At the end of the experiment, blood samples were taken and then the sheep were slaughtered and tissue samples from the rumen ventral sac, rumen dorsal sac, liver, duodenum and Longissimus dorsi muscle were obtained. IGF-I, IGFBP-3, GH, insulin and glucagon in plasma and IGF-I and IGFBP-3 in different tissues were analysed. Results showed that the concentration of IGF-I, IGFBP-3, GH, insulin or glucagon in plasma and the content of IGF-I and IGFBP-3 in the rumen dorsal sac, rumen ventral sac, liver or Longissimus dorsi muscle were increased with VFA infusion level (p<0.05). No significant differences were found in duodenum IGF-I between Treatments I and II and in rumen dorsal sac IGFBP-3 between Treatments II and III (p>0.05). It was concluded that IGF-I, IGFBP-3, GH, insulin and glucagon in plasma and IGF-I and IGFBP-3 in rumen dorsal sac, rumen ventral sac, liver and Longissimus dorsi muscle were increased significantly with increasing level of ruminal infusion of mixed VFA.

Clinical Safety and Effectiveness of Stent-Assisted Coil Embolization with Neuroform Atlas Stent in Intracranial Aneurysm

  • Kim, Chang Hyeun;Kim, Young Ha;Sung, Soon Ki;Son, Dong Wuk;Song, Geun Sung;Lee, Sang Weon
    • Journal of Korean Neurosurgical Society
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    • v.63 no.1
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    • pp.80-88
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    • 2020
  • Objective : Stent-assisted coil embolization (SAC) is commonly used for treating wide-neck intracranial aneurysms. In this study, we aimed to assess the clinical safety and efficacy of the NeuroForm Atlas Stent during SAC of intracranial aneurysms. Methods : We retrospectively analyzed data from patients with ruptured and unruptured cerebral aneurysms, who underwent SAC using the NeuroForm Atlas between February 2018 and July 2018. Favorable clinical outcomes and degree of aneurysm occlusion were defined as a modified Rankin scale score of ≤2 and a Raymond-Roy occlusion classification (RROC) class I/II during the immediate postoperative period and at the 6-month follow-up, respectively. Results : Thirty-one consecutive patients with 33 cases, including 11 ruptured and 22 unruptured cases were treated via NeuroForm Atlas SAC. Among the 22 unruptured cases with 24 unruptured aneurysms had favorable clinical outcome. Complete occlusion (RROC I) was achieved in 16 aneurysms (66.7%), while neck remnants (RROC II) were observed in six aneurysms (25%). Among the 11 patients with ruptured aneurysms, two died due to re-bleeding and diabetic ketoacidosis. In ruptured cases, RROC I was observed in eight (72.7%) and RROC II was observed in three cases (27.3%). At the 6-month follow-up, no clinical events were observed in the 22 unruptured cases. In the ruptured nine cases, five patients recovered without neurologic deficits, while four experienced unfavorable outcomes at 6 months. Of the 29 aneurysms examined via angiography at the 6-month follow-up, 19 (65.5%) were RROC I, eight (27.6%) were RROC II and two (6.9%) were RROC III. There were no procedure-related hemorrhagic complications. Conclusion : In this study, we found that stent-assisted coil embolization with NeuroForm Atlas stent may be safe and effective in the treatment of wide-neck intracranial aneurysms. NeuroForm Atlas SAC is feasible for the treatment of both ruptured and unruptured wide-neck aneurysms.

Type 1.5 Split Cord Malformation : A New Theory of Pathogenesis

  • Sun, Mengchun;Tao, Benzhang;Luo, Tianbao;Gao, Gan;Shang, Aijia
    • Journal of Korean Neurosurgical Society
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    • v.65 no.1
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    • pp.138-144
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    • 2022
  • To report two cases of type 1.5 split cord malformation (SCM), a subtype of SCM with combined characteristics of types I and II and to review the relevant literature and propose a new possible pathogenetic theory for type 1.5 SCM. A 52-year-old woman had hemicords within a single dural sac with a dorsal bony septum at the L5 level. A 9-year-old boy had hemicords within a single dural sac with a ventral bony septum and fibrous extension at the L3 level. Both patients underwent microsurgical treatments for removing the bony septum, detethering the spinal cord, and sectioning the filum terminale. The surgical procedure revealed an extradural partial bony septum and hemicords within an intact single dural sac in each patient. Both patients were discharged from the hospital without de novo nerve dysfunction. Published cases have validated that types I and II SCM can overlap. We recommend recent type 1.5 SCM as a normative terminology for this overlapping SCM and report two rare cases of this SCM. We propose an associated pathogenesis consisting of uneven distribution and regression to explain type 1.5 SCM. Furthermore, we postulate that the amount of condensing meninx primitiva might determine whether the left bony septum has fibrous extensions to the opposite dura in type 1.5 SCM.

Analysis of the Relationship between MHC-DRB1 Gene Polymorphism and Hydatidosis in Kazakh Sheep

  • Li, Ren-Yan;Jia, Bin;Zhang, Wen-Ju;Zhao, Zong-Sheng;Shi, Guo-Qing;Shen, Hong;Peng, Qiang;Lv, Li-Min;Zhou, Qi-Wei;Du, Ying-Chun
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1145-1151
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    • 2010
  • The objective of this work was to analyze the relationship between ovine major histocompatibility complex (MHC) DRB1 gene polymorphism and genetic resistance to hydatidosis in Kazakh sheep. The Ovar (ovine MHC) class II DRB1 second exon was amplified by polymerase chain reaction (PCR) from DNA samples of 702 Kazakh sheep, including 302 sheep with hydatidosis and 400 health controls. PCR products were characterized by the restriction fragment length polymorphism (RFLP) technique using five restriction enzymes, i.e., MvaI, HaeIII, SacI, SacII and Hin1I, yielding 14 alleles and 28 genotypes. Comparing the frequency of genotypes in hydatidosis sheep with the control group, it was found that the genotype frequencies of MvaIbc, Hin1Iab, SacIIab, HaeIIIde, HaeIIIdf and HaeIIIdd in control sheep were significantly (p<0.01) higher than in hydatidosis sheep, indicating that a significant correlation existed between these genotypes and resistance to hydatidosis. Genotype frequencies of MvaIbb, SacIIaa, Hin1Ibb and HaeIIIef in sheep with hydatidosis were extremely significantly (p<0.01) higher than in the control group, and the genotype frequency of HaeIIIab was significantly higher (p<0.05), indicating that a marked correlation existed between these genotypes and susceptibility to hydatidosis. By way of analyzing haplotype with these resistant genotypes, the hydatidosis resistant haplotype MvaIbc-SacIIab-Hin1Iab of Kazakh sheep was screened out, and then verified through artificial hydatid infection in sheep. The results indicated that the infection rate of sheep with the resistant haplotype of hydatidosis was significantly lower (p<0.01) than without this resistant haplotype. It showed that the genic haplotype MvaIbc-SacIIab-Hin1Iab of Ovar-DRB1 exon 2 was the resistant haplotype of hydatidosis in Kazakh sheep.

A Study on the Absorption Enhancing Effect of Sodium 5-Methoxysalicylate(II) (Sodium 5-Methoxy Salicylate의 흡수촉진 효과에 관한 연구 (II). 흡수촉진제에 대한 반응성)

  • 김기헌
    • YAKHAK HOEJI
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    • v.30 no.2
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    • pp.96-99
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    • 1986
  • Absorption enhancing effect of 5 MSA-Na at the mucous memberanes of the rectum and duodenum was studied via in vitro sac test. There were no significant differences between two membranes. Effect of 5MSA-Na on the transfer of CMZ to the tissue was also studied via in situ loop method. There was no significant difference at the duodenum, compared to the control when CMZ was administered intravenously. Transfer of CMZ to the loop of the rectum was increased in the presence of 5MSA-Na compared to the control, which might be attributed to the enhanced permeability of the mucous memberane of the rectum.

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Effect of Bupleurum falcatum on the immune system (시호 추출물이 면역계 세포의 활성에 미치는 영향)

  • Cho, Jeong-gon;Kim, Jong-myeon
    • Korean Journal of Veterinary Research
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    • v.34 no.4
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    • pp.769-779
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    • 1994
  • The root of Bupleurum falcatum L.(BF) has been widely used in oriental medicine as a major camponent in many prescriptions for chronic hepatitis, renal disease, tuberculosis and some other infectious diseases. Many attempts have done to investigate the therapeutic effects of these principles. However, any kinds of screenig on immune regulatory- and antitumor- effects of BF has not been reported. The present study, therefore, was undertaken to investigate the BF-effects on cellular- and humoral-immune responses, phagocytic activities of macrophages, lymphokine- and Immunoglobulin(Ig)-production of lymphocytes, tumorigenesis of implanted sarcoma 180 cells and B16 melanoma cells, and proliferations of some tumor cell lines(Fsa II, 3LL and EL4). BF increased phagocytic activities of mouse peritoneal macrophages in a dose- and time-dependent fashion. Arthus reaction and antibody responses to SRBC were slightly enhanced but delayed hypersensitivity was depresed when BF was injected before- and after-SRBC sensitization. BF inhibited the proliferative responses of human tonsillar lymphocytes to PHA- and Con A-stimulation but slightly augmented the response of these cells to Staphylococcus aureus Cowan 1(SAC)-activation. Ig secretion of human mononuclear cells activated with SAC was slightly increased by BF. BF significantly augmented the SAC-induced IL 6 production of human mononuclear cells but not influenced Con Ainduced IL 2 secretion. NK cell activities of mouse splenocytes were somewhat increased when BF was pretreated and this responses were due to the increment of binding affinities of effector cells to target cells and of lytic activities of effector cells against target cells. In vitro BF significantly inhibited the proliferations of cancer cells such as Fsa II, 3LL and EL4 strains. BF decreased not only the frequency of tumor induction but also the tumor size per sarcoma 180 or B16 cell-implanted mouse. Taken together, these results indicate that BF is one of the potential immunomodulator, and suggest its possibility to be used as a desirable antitumor agent.

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Application of Screening Technology for Capture of Hydrogen Sulfide Using Ionic Liquids (이온성 액체의 황화수소의 포집을 위한 스크리닝 기법의 활용)

  • Han, Sangil;Lee, Bong-Seop
    • Journal of Industrial Technology
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    • v.39 no.1
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    • pp.41-45
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    • 2019
  • Hydrogen sulfide ($H_2S$) is mainly produced along with methane and hydrocarbons in many gas fields as well as hydrodesulfurization processes of crude oils containing sulfur compounds and the emission of $H_2S$ has a considerable effect on both environmental problem and human health aspects due to formation of, e.g. acid rain and smog. In recent years, ionic liquids (ILs) have been proposed as the most promising solvents for $CO_2$ and hazardous pollutants capture, such as $H_2S$ and sulfur dioxide ($SO_2$). In this work, we demonstrate the use of the predictive COSMO-SAC model for the prediction of Henry's law constant of $H_2S$ in ILs. Furthermore, the method is used to screen for potential IL candidates for $H_2S$ capture from a set of 2,624 ILs formed from 82 cations and 32 anions. The effects of cation on the Henry's law constant of $H_2S$ such as (i) the variation of the alkyl chain length on cation, (ii) the substituent of methyl group ($-CH_3$) for H in C(2) position and (iii) the change of ring structure for cation family are clearly predicted by COSMO-SAC model.