• Title/Summary/Keyword: Extracellular

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Purification of Extracellular Enzyme Produced by Vibrio sp. AL-145 (Vibrio sp. AL-145가 생산하는 균체외 효소의 정제 (I))

  • 주동식;이응호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.2
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    • pp.234-239
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    • 1993
  • The alginate degrading bacteria have been screened from the marine environment. Sodium alginate and NaCl were required for cell growth and enzyme production of 145-C strain and the adequate concentrations were 0.7 and 2.5%, respectively. The effective nitrogen source was peptone and adequate temperature was 28$\pm$2$^{\circ}C$. The 145-C strain was identified as Vibrio sp. from biochemical and biological experiment. The extracellular enzyme produced by Vibrio sp. was purified and the molecular weight was estimated to be 27, 000.

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Therapeutic application of extracellular vesicles for various kidney diseases: a brief review

  • Lee, Sul A;Yoo, Tae Hyun
    • BMB Reports
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    • v.55 no.1
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    • pp.3-10
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    • 2022
  • Extracellular vesicles (EVs) released from different types of kidney cells under physiologic conditions contribute to homeostasis maintenance, immune-modulation, and cell-to-cell communications. EVs can also negatively affect the progression of renal diseases through their pro-inflammatory, pro-fibrotic, and tumorigenic potential. Inhibiting EVs by blocking their production, release, and uptake has been suggested as a potential therapeutic mechanism based on the significant implication of exosomes in various renal diseases. On the other hand, stem cell-derived EVs can ameliorate tissue injury and mediate tissue repair by ameliorating apoptosis, inflammation, and fibrosis while promoting angiogenesis and tubular cell proliferation. Recent advancement in biomedical engineering technique has made it feasible to modulate the composition of exosomes with diverse biologic functions, making EV one of the most popular drug delivery tools. The objective of this review was to provide updates of recent clinical and experimental findings on the therapeutic potential of EVs in renal diseases and discuss the clinical applicability of EVs in various renal diseases.

Extracellular Vesicles as an Endocrine Mechanism Connecting Distant Cells

  • Kita, Shunbun;Shimomura, Iichiro
    • Molecules and Cells
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    • v.45 no.11
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    • pp.771-780
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    • 2022
  • The field of extracellular vesicles (EVs) has expanded tremendously over the last decade. The role of cell-to-cell communication in neighboring or distant cells has been increasingly ascribed to EVs generated by various cells. Initially, EVs were thought to a means of cellular debris or disposal system of unwanted cellular materials that provided an alternative to autolysis in lysosomes. Intercellular exchange of information has been considered to be achieved by well-known systems such as hormones, cytokines, and nervous networks. However, most research in this field has searched for and found evidence to support paracrine or endocrine roles of EV, which inevitably leads to a new concept that EVs are synthesized to achieve their paracrine or endocrine purposes. Here, we attempted to verify the endocrine role of EV production and their contents, such as RNAs and bioactive proteins, from the regulation of biogenesis, secretion, and action mechanisms while discussing the current technical limitations. It will also be important to discuss how blood EV concentrations are regulated as if EVs are humoral endocrine machinery.

The complex role of extracellular vesicles in HIV infection

  • Jung-Hyun Lee
    • BMB Reports
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    • v.56 no.6
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    • pp.335-340
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    • 2023
  • During normal physiological and abnormal pathophysiological conditions, all cells release membrane vesicles, termed extracellular vesicles (EVs). Growing evidence has revealed that EVs act as important messengers in intercellular communication. EVs play emerging roles in cellular responses and the modulation of immune responses during virus infection. EVs contribute to triggering antiviral responses to restrict virus infection and replication. Conversely, the role of EVs in the facilitation of virus spread and pathogenesis has been widely documented. Depending on the cell of origin, EVs carry effector functions from one cell to the other by horizontal transfer of their bioactive cargoes, including DNA, RNA, proteins, lipids, and metabolites. The diverse constituents of EVs can reflect the altered states of cells or tissues during virus infection, thereby offering a diagnostic readout. The exchanges of cellular and/or viral components by EVs can inform the therapeutic potential of EVs for infectious diseases. This review discusses recent advances of EVs to explore the complex roles of EVs during virus infection and their therapeutic potential, focusing on HIV-1.

Effects of Extracellular $Ca^{++}$ on PKC or cAMP-stimulated Increases in LH Release and $LH{\beta}$ Subunit mRNA Levels in Rat Anterior Pituitary Cells (흰쥐 뇌하수체 전엽세포에서 PKC나 cAMP에 의한 LH 분비 및 $LH{\beta}$ Subunit mRNA 증가에 미치는 $Ca^{++}$의 영향)

  • Park, Deok-Bae;Kim, Chang-Mee;Cheon, Min-Seok;Ryu, Kyung-Za
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.347-355
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    • 1996
  • We examined the effects of EGTA and verapamil on phorbol ester-and forskolin-stimulated LH releases and $LH{\beta}$ subunit mRNA levels in order to verify the role of extracellular $Ca^{++}$ on PKC- or cAMP-induced increases in LH release and $LH{\beta}$ subunit mRNA levels in cultured anterior pituitary cells of rat. Forskolin-stimulated $LH{\beta}$ subunit mRNA levels as well as LH release were all suppressed by prevention of $Ca^{++}$ mobilization from extracellular environment, after the treatment of EGTA as a $Ca^{++}$ chelator or verapamil as a $Ca^{++}$ channel blocker. PMA-stimulated $LH{\beta}$ subunit mRNA levels were also suppressed by the treatment of EGTA and verapamil, while PMA-induced LH release was not affected. From the present study, it is, therefore, suggested that PKC activation and cAMP elevation all stimulate $LH{\beta}$ subunit mRNA levels and these are extracellular $Ca^{++}$-dependent. However, LH releases by PKC activation and cAMP increase seem to be different each other. LH release by PKC activation is thought to be independent of extracellular $Ca^{++}$. On the other hand, cAMP stimulated-LH release is thought to be dependent on the entry of extracellular $Ca^{++}$.

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Distribution, Biovolume and Extracellular Enzyme Activities of Heterotrophic Bacteria in the Sea near Kunsan,Korea (군산 인근 해역에서 종속영양 세균의 분포, 평균체적 및 세포외 효소활성력)

  • 이건형;김재원;김정희
    • The Korean Journal of Ecology
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    • v.17 no.1
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    • pp.79-90
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    • 1994
  • Water samples were taken at 6 stations from the mouth of Keum River to Kogunsan Archipelago of West Sea during December, 1991 to August, 1992, to determine the distribution of heterotrophic bacteria, their biovolumes and heterotrophic activities. Heterotrophic marine bacteria ranged from $1.0\;{\times}\;10^3to\;5\;{\times}\;10^5c.f.u.$ /ml. As for morphological distribution measured by epifluorescence microscopy, rod-shaped bacteria were between 45% and 72% of all cells during investigation period. Average biovolume of sampled bacteria ranged from $(7.69\;{\pm}\;0.18)\;{\times}10^{-2}to\;(8.18\;{\pm}\;0.38)\;{\times}\;10^{-2}\;{\mu}m^3$ for coccoid bacteria, and from $(6.09\;{\pm}\;0.29)\;{\times}10^{-2}to\;(7.72\;{\pm}\;0.41)\;{\times}\;10^{-2}\;{\mu}m^3$ for rod-shaped ones. The activities of extracellular enzymes ranged from 0.01 to 2.6 ${\mu}M$ /l /hr for glucosidase, from 0.01 to 2.6 ${\mu}M$ /l /hr for amylase, from 0.01 to 8.86 ${\mu}M$ /I /hr phosphatase and from 0.01 to 0.94 ${\mu}M$ /l /hr for chitinase. Extracellular enzyme activities were higher in summer season than in other sampling periods, and phosphatase showed the highest activity among measured extracellular enzymes. Bacterial distribution and their extracellular enzyme activities were associated with water temperature and organic nutrients, but bacterial cell volumes showed no direct relationship with extracellular enzyme activities.

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Production of Extracellular Lipid by Rhodotorula graminis SW 214 (Rhodotorula graminis SW 214의 세포외 지방질 생산에 관한 연구)

  • Kim, Sung-Yeun;Kough, Kyoung;Park, Sung-Oh
    • Korean Journal of Food Science and Technology
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    • v.25 no.6
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    • pp.794-800
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    • 1993
  • A lipid producing yeast was screened from leaves of Albabiscus and was identified as a Rhodotorula graminis SW 214. During the shakining incubation of 8 days at $25^{\circ}C$, the yeast produced extracellular lipids of 7.3g/l of the media. The relative concentration of carbon and nitrogen sources in the media influenced the extracellular lipid production greatly. When with nitrogen sources in the media were almost exhausted for growth of the yeast the sufficient carbon sources, the lipid production proceeded vigorously. Eight days of batch cultivation with 8% glucose, 2.5g/l of yeast extract, $KH_{2}PO_{4}(1g/l)\;MgSO_{4}\;(0.2g/l)$ and pH 6 gave maximum biomass and extracellular lipid production of 8.05g/l and 8.89g/l, respectively. The acid value, saponificatio value, the iodine value, ad the unsaponifiable matter of the extracellular lipids of Rhodotorula graminis SW 214 were 2.6, 534, 5.1 and 2.4, respectively. Lipid was constituted 75.2% triglyceride, 5.9% free fatty acid, 10.8% phospholipid, 4.9% esterified sterol and 3.3% free sterol. Major fatty acids found were 3-hydroxypentadecanoate, 3-hydroxyhexadecanate, trans-9-octadecanate, cis-9-hexadecanate (hydroxy palmitic), 15-methylhexadecanate (oleic), 18-methylno-nadecanate, octadecanate (stearic) and 3-hydroxytridecante.

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Effect of Glucose at High Concentrations on the Apoptosis of the Cultured Periodontal Ligament Cells and Osteoblasts (고농도 포도당이 뼈모세포와 치주인대세포의 세포자멸사에 미치는 영향에 관한 연구)

  • Park, Sung-Ho;Jue, Seong-Suk;Hong, Jung-Pyo;Shin, Je-Won
    • Journal of Oral Medicine and Pain
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    • v.32 no.4
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    • pp.357-364
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    • 2007
  • This experiment was designed to clarify the effect of extracellular glucose on the osteoblasts and periodontal ligament cells. The cells were incubated for 24 and 48 hours with ${\alpha}$-MEM including 1,000 mg/L (control group) and 4,500 mg/L (experimental group) of glucose. Then, the expressions of caspase-3, p38 MAPK, JNK-1, and ERK-1 were examined using Elisa assay and Western blot. The results were as follows: 1. The expression of caspase-3 and p38 MAPK was increased by the high extracellular glucose in both cells. 2. The expression of caspase-3 and p38 MAPK was increased greatly in the periodontal ligament cells than the E1 cells by the high extracellular glucose. 3. The expression of JNK-1 was increased by the high extracellular glucose in both cells. 4. The expression of ERK-1 was not changed by the high extracellular glucose in both cells. These results suggest that extracellular glucose at high concentrations may inhibit the periodontal regeneration process increasing cellular apoptosis. And p38 MAPK and JNK-1 pathway may be the most responsible intracellular pathway rather than ERK-1.

Characterization of Sporulation-Specific Glucoamylase of Saccharomyces diastaticus (Saccharomyces diastaticus의 포자형성 특이 글루코아밀라제의 특성)

  • Kim, Eun-Ju;Ahn, Jong-Seog;Kang, Dae-Ook
    • Journal of Life Science
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    • v.20 no.5
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    • pp.683-690
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    • 2010
  • The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

Purification and Characterization of Caseinolytic Extracellular pretense from Bacillus amyloliquefaciens S94

  • Son, Eui-Sun;Kim, Jong-Il
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.26-32
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    • 2002
  • From the culture supernatant of the psychrotrophic strain of Bacillus amyloliquefaciens an extracellular serine protease was purified to apparent homogeneity by successive purification steps using QAE-Sephadex, SP-Sephadex and Sephacryl S-100 column chromatography. The pretense is monomeric, with a relative molecular mass of 23,000. It is inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride, but not by EDTA. The enzyme is most active at pH 9-10 and at $45^{\circ}C$, although it is unstable at $60^{\circ}C$.