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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Microbiology and Biotechnology
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 18, Issue 12 - Dec 2008
Volume 18, Issue 11 - Nov 2008
Volume 18, Issue 10 - Oct 2008
Volume 18, Issue 9 - Sep 2008
Volume 18, Issue 8 - Aug 2008
Volume 18, Issue 7 - Jul 2008
Volume 18, Issue 6 - Jun 2008
Volume 18, Issue 5 - May 2008
Volume 18, Issue 4 - Apr 2008
Volume 18, Issue 3 - Mar 2008
Volume 18, Issue 2 - Feb 2008
Volume 18, Issue 1 - Jan 2008
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Control of Tylosin Biosynthesis in Streptomyces fradiae
Cundliffe, Eric ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1485~1491
Tylosin biosynthesis is controlled in cascade fashion by multiple transcriptional regulators, acting positively or negatively, in conjunction with a signalling ligand that acts as a classical inducer. The roles of regulatory gene products have been characterized by a combination of gene expression analysis and fermentation studies, using engineered strains of S. fradiae in which specific genes were inactivated or overexpressed. Among various novel features of the regulatory model, involvement of the signalling ligand is not essential for tylosin biosynthesis.
Specific Detection of Xanthomonas oryzae pv. oryzicola in Infected Rice Plant by Use of PCR Assay Targeting a Membrane Fusion Protein Gene
Kang, Man-Jung ; Shim, Jae-Kyung ; Cho, Min-Seok ; Seol, Young-Joo ; Hahn, Jang-Ho ; Hwang, Duk-Ju ; Park, Dong-Suk ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1492~1495
Successful control of Xanthomonas oryzae pv. oryzicola, the causal agent of bacterial leaf streak, requires a specific and reliable diagnostic tool. A pathovar-specific PCR assay was developed for the rapid and accurate detection ofthe plant pathogenic bacterium Xanthomonas oryzae pv. oryzicola in diseased plant. Based on differences in a membrane fusion protein gene of Xanthomonas oryzae pv. oryzicola and other microorganisms, which was generated from NCBI (http://www.ncbi.nlm.nih.gov/) and CMR (http://cmr.tigr.org/) BLAST searches, one pair of pathovar-specific primers, XOCMF/XOCMR, was synthesized. Primers XOCMF and XOCMR from a membrane fusion protein gene were used to amplity a 488-bp DNA fragment. The PCR product was only produced from 4 isolates of Xanthomonas oryzae pv. oryzicola among 37 isolates of other pathovars and species of Xanthomonas, Pectobacterium, Pseudomonas, Burkholderia, Escherichia coli, and Fusarium oxysporum f.sp. dianthi. The results suggested that the assay detected the pathogen more rapidly and accurately than standard isolation methods.
Lysobacter ginsengisoli sp. nov., a Novel Species Isolated from Soil in Pocheon Province, South Korea
Jung, Hae-Min ; Ten, Leonid N. ; Im, Wan-Taek ; Yoo, Soon-Ae ; Lee, Sung-Taik ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1496~1499
A Gram-negative, aerobic, rod-shaped, nonspore-forming bacterial strain, designated Gsoil
was isolated from soil sample of a ginseng field in Pocheon Province (South Korea). The isolate contained Q-8 as the predominant ubiquinone and iso-
, and iso-
as the major fatty acids. The G+C content of the genomic DNA was 69.3 mol%. A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Gsoil
was most closely related to Lysobacter gummosus (97.6%) and Lysobacter antibioticus (97.6%). However, the DNA-DNA relatedness value between strain Gsoil
and its phylogenetically closest neighbors was less than 17%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil 357T should be classified as representing a novel species in the genus Lysobacter, for which the name Lysobacter ginsengisoli sp. novo is proposed. The type strain is Gsoil
Avirulence Gene Diversity of Xanthomonas axonopodis pv. glycines Isolated in Korea
Park, Hyoung-Joon ; Han, Sang-Wook ; Oh, Chang-Sik ; Lee, Seung-Don ; Ra, Dong-Soo ; Lee, Suk-Ha ; Heu, Sung-Gi ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1500~1509
The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean cultivars collected in Korea and were classified into six different type strains of OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 according to the patterns of avrBs3-homologous bands. When these type strains were inoculated on various cultivars, most of the Korean strains mildly induced disease symptoms on the resistant CNS1 cultivars. Unlike other type strains, strain SL2098, which appeared not to contain any avrBs3 homolog, induced only a few pustules on even highly susceptible cultivars. When a plasmid carrying the 3.7-kb avrBs3-homologous gene from strain SL1045 was introduced into SL2098, the transformant could not recover the pathogenicity in susceptible host plants. However, when avrBs3-homologous genes of strain SL1018 were mutated by transposon mutagenesis, one of the mutants in which a 5.2-kb chromosomal band homologous to avrBs3 was disrupted could not induce the hypersensitive response on resistant cultivars such as William82 or CNS2. Our results suggest that the avrBs3 homologs may play important roles in the pathogenicity of Xanthomonas axonopodis pv. glycines and the recognition of soybean cultivars.
Generation of Expressed Sequence Tags for Immune Gene Discovery and Marker Development in the Sea Squirt, Halocynthia roretzi
Kim, Young-Ok ; Cho, Hyun-Kook ; Park, Eun-Mi ; Nam, Bo-Hye ; Hur, Young-Baek ; Lee, Sang-Jun ; Cheong, Jae-Hun ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1510~1517
Expresssed sequence tag (EST) analysis was developed from three cDNA libraries constructed from cells of the digestive tract, gonad, and liver of sea squirt. Randomly selected cDNA clones were partially sequenced to generate a total of 922 ESTs, in which 687 unique ESTs were identified respectively. Results of BLASTX search showed that 612 ESTs (89%) have homology to genes of known function whereas 75 ESTs (11%) were unidentified or novel. Based on the major function of their encoded proteins, the identified clones were classified into ten broad categories. We also identified several kinds of immune-related genes as identifying novel genes. Sequence analysis of ESTs revealed the presence of microsatellite-containing genes that may be valuable for further gene mapping studies. The accumulation of a large number of identified cDNA clones is invaluable for the study of sea squirt genetics and developmental biology. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.
The Role of AiiA, a Quorum-Quenching Enzyme from Bacillus thuringiensis, on the Rhizosphere Competence
Park, Su-Jin ; Park, Sun-Yang ; Ryu, Choong-Min ; Park, Seung-Hwan ; Lee, Jung-Kee ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1518~1521
Bacteria sense their population density and coordinate the expression of target genes, including virulence factors in Gram-negative bacteria, by the N-acylhomoserine lactones (AHLs)-dependent quorum sensing (QS) mechanism. In contrast, several soil bacteria are able to interfere with QS by enzymatic degradation of AHLs, referred to as quorum quenching. A potent AHL-degrading enzyme, AiiA, from Bacillus thuringiensis has been reported to effectively attenuate the virulence of bacteria by quorum quenching. However, little is known about the role of AiiA in B. thuringiensis itself. In the present study, an aiiA-defective mutant was generated to investigate the role of AHA in rhizosphere competence in the root system of pepper. The aiiA mutant showed no detectable AHL￢-egrading activity and was less effective for suppression of soft-rot symptom caused by Erwinia carotovora on the potato slice. On the pepper root, the survival rate of the aiiA mutant significantly decreased over time compared with that of wild type. Interestingly, viable cell count analysis revealed that the bacterial number and composition of E. carotovora were not different between treatments of wild type and the aiiA mutant. These results provide evidence that AHA can play an important role in rhizosphere competentce of B. thuringiensis and bacterial quorum quenching to Gram-negative bacteria without changing bacterial number or composition.
Morphogenetic Behavior of Tropical Marine Yeast Yarrowia lipolytica in Response to Hydrophobic Substrates
Zinjarde, Smita S. ; Kale, Bhagyashree V. ; Vishwasrao, Paresh V. ; Kumar, Ameeta R. ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1522~1528
The morphogenetic behavior of a tropical marine Yarrowia lipolytica strain on hydrophobic substrates was studied. Media containing coconut oil or palm kernel oil (rich in lauric and myristic acids) prepared in distilled water or seawater at a neutral pH supported 95% of the cells to undergo a transition from the yeast form to the mycelium form. With potassium laurate, 51 % of the cells were in the mycelium form, whereas with myristate, 32% were in the mycelium form. However, combinations of these two fatty acids in proportions that are present in coconut oil or palm kernel oil enhanced the mycelium formation to 65%. The culture also produced extracellular lipases during the morphogenetic change. The yeast cells were found to attach to the large droplets of the hydrophobic substrates during the transition, while the mycelia were associated with the aqueous phase. The alkane-grown yeast partitioned more efficiently in the hydrophobic phases when compared with the coconut oil-grown mycelia. A fatty acid analysis of the mycelial form revealed the presence of lauric acid in addition to the long-chain saturated and unsaturated fatty acids observed in the yeast form. The mycelia underwent a rapid transition to the yeast form with n-dodecane, a medium-chain aliphatic hydrocarbon. Thus, the fungus displayed a differential behavior towards the two types of saturated hydrophobic substrates.
The Diversity of Lysine-Acetylated Proteins in Escherichia coli
Yu, Byung-Jo ; Kim, Jung-Ae ; Moon, Jeong-Hee ; Ryu, Seong-Eon ; Pan, Jae-Gu ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1529~1536
Acetylation of lysine residues in proteins is a reversible and highly regulated posttranslational modification. However, it has not been systematically studied in prokaryotes. By affinity immunoseparation using an anti-acetyllysine antibody together with nano-HPLC/MS/MS, we identified 125 lysine-acetylated sites in 85 proteins among proteins derived from Escherichia coli. The lysine-acetylated proteins identified are involved in diverse cellular functions including protein synthesis, carbohydrate metabolism, the TCA cycle, nucleotide and amino acid metabolism, chaperones, and transcription. Interestingly, we found a higher level of acetylation during the stationary phase than in the exponential phase; proteins acetylated during the stationary phase were immediately deacetylated when the cells were transferred to fresh LB culture medium. These results demonstrate that lysine acetylation is abundant in E. coli and might be involved in modifying or regulating the activities of various enzymes involved in critical metabolic processes and the synthesis of building blocks in response to environmental changes.
Amendment with Peony Root Bark Improves the Biocontrol Efficacy of Trichoderma harzianum against Rhizoctonia solani
Lee, Tae-Ok ; Khan, Zakaullah ; Kim, Sang-Gyu ; Kim, Young-Ho ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1537~1543
We tested Trichoderma harzianum as a biocontrol agent for Rhizoctonia solani AG2-1, using six natural antifungal materials to improve its efficacy. Among the six materials tested, peony (Paeonia suffruticosa) root bark (PRB) showed the strongest antifungal activity against R. solani AG2-1, and was not antagonistic to T. harzianum. Scanning electron microscopy showed that treatment with PRB extract resulted in shortened and deformed R. so/ani AG2-1 hyphal cells. The control of radish damping-off caused by R. so/ani AG2-1 was greatly increased by combined treatments of T. harzianum and PRB, as compared with either of the two treatments alone, with the control effect increased from 42.3-51.5% to 71.4-87.6%. The antifungal compound in PRB, which was isolated in chloroform and identified as paeonol by mass spectrometry,
NMR analyses, inhibited the growth of R. so/ani AG2-1 but not that of T. harzianum. Thus, PRB powder or extract may be used as a safe additive to T. harzianum to improve the control of the soil borne diseases caused by R. so/ani AG2-1.
Bifunctional Recombinant Fusion Enzyme Between Maltooligosyltrehalose Synthase and Maltooligosyltrehalose Trehalohydrolase of Thermophilic Microorganism Metallosphaera hakonensis
Seo, Ju-Seok ; An, Ju-Hee ; Cheong, Jong-Joo ; Choi, Yang-Do ; Kim, Chung-Ho ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1544~1549
MhMTS and MhMTH are trehalose (
-D-glucopyranose) biosynthesis genes of the thermophilic microorganism Metallosphaera hakonensis, and encode a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively. In this study, the two genes were fused in-frame in a recombinant DNA, and expressed in Escherichia coli to produce a bifunctional fusion enzyme, MhMTSH. Similar to the two-step reactions with MhMTS and MhMTH, the fusion enzyme catalyzed the sequential reactions on maltopentaose, maltotriosyltrehalose formation, and following hydrolysis, producing trehalose and maltotriose. Optimum conditions for the fusion enzyme-catalyzed trehalose synthesis were around
and pH 5.0-6.0. The MhMTSH fusion enzyme exhibited a high degree of thermostability, retaining 80% of the activity when pre-incubated at
for 48 h. The stability was gradually abolished by incubating the fusion enzyme at above
. The MhMTSH fusion enzyme was active on various sizes of maltooligosaccharides, extending its substrate specificity to soluble starch, the most abundant natural source of trehalose production.
Development of a Method to Measure Hydrogen Sulfide in Wine Fermentation
Park, Seung-Kook ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1550~1554
A hydrogen sulfide
detecting tube was developed for the quantitative determination of
produced by yeast during laboratory scale wine fermentations. The detecting tube consisted of a small transparent plastic tube packed with an
-sensitive color-indicating medium. The packed medium changed color, with the color change progressing upward from the bottom of the tube, upon exposure to
produced by yeast during fermentation. A calibration study using a standard
gas showed that the length of the portion that darkened was directly related to the quantity of
) with a high correlation coefficient (
=0.9997). The reproducibility of the
detecting tubes was determined with five repetitive measurements using a standard
/200 ml (28 ppb)], which resulted in a coefficient of variation of 3.6% at this level of
. With the sulfide detecting tubes, the production of
was continuously monitored and quantified from laboratory scale wine fermentations with different yeast strains and with the addition of different levels of elemental sulfur to the grape juice. This sulfide detecting tube technology may allow winemakers to quantitatively measure
produced under different fermentation conditions, which will eventually lead winemakers to better understand the specific factors and conditions for the excessive production of
during wine fermentation in a large production scale.
Purification, Characterization, and Partial Primary Sequence of a Major-Maltotriose-producing
-Amylase, ScAmy43, from Sclerotinia sclerotiorum
Ben Abdelmalek-Khedher, Imen ; Urdad, Maria Camino ; Limam, Ferid ; Schmitter, Jean Marie ; Marzouki, M. Nejib ; Bressollier, Philippe ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1555~1563
-D-glucan glucanohydrolase, E.C. 184.108.40.206), ScAmy43, was found in the culture medium of the phytopathogenic fungus Sclerotinia sclerotiorum grown on oats flour. Purified to homogeneity, ScAmy43 appeared as a 43 kDa monomeric enzyme, as estimated by SDS-PAGE and Superdex 75 gel filtration. The MALDI peptide mass fingerprint of ScAmy43 tryptic digest as well as internal sequence analyses indicate that the enzyme has an original primary structure when compared with other fungal a-amylases. However, the sequence of the 12 N-terminal residues is homologous with those of Aspergillus awamori and Aspergillus kawachii amylases, suggesting that the new enzyme belongs to the same GH13 glycosyl hydrolase family. Assayed with soluble starch as substrate, this enzyme displayed optimal activity at pH 4 and
with an apparent
value of 1.66 mg/ml and
. ScAmy43 activity was strongly inhibited by
, moderately by
, and was only weakly affected by
addition. However, since EDTA and EGTA did not inhibit ScAmy43 activity, this enzyme is probably not a metalloprotein. DTT and
-mercaptoethanol strongly increased the enzyme activity. Starting with soluble starch as substrate, the end products were mainly maltotriose, suggesting for this enzyme an endo action.
Reductive Dechlorination of Low Concentration Polychlorinated Biphenyls as Affected by a Rhamnolipid Biosurfactant
Kim, Jong-Seol ; Frohnhoefer, Robert C. ; Cho, Young-Cheol ; Cho, Du-Wan ; Rhee, G-Yull ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1564~1571
We investigated whether the threshold concentration for polychlorinated biphenyl (PCB) dechlorination may be lower in biosurfactant-amended sediments compared with biosurfactant-free samples. At PCB concentrations of 40, 60, and 120 ppm, the surfactant amendment enhanced the PCB dechlorination rate at all concentrations and the rate was also faster at higher concentrations. On a congener group basis, dechlorination proceeded largely with group A (congeners with low threshold) in both surfactant-free and -amended sediments, accumulating mainly group C (residual products of dechlorination) congeners, and surfactant enhanced the dechlorination rate of group A congeners. Since the PCB threshold concentration for the inoculum in the experiment was lower than 40 ppm, we carried out another experiment using sediments with lower PCB concentrations, 10, 20, and 30 ppm. Sediments with 100 ppm were also performed to measure dechlorination at a PCB saturation concentration. Comparison between the plateaus exhibited that the extent of dechlorination below 40 ppm PCBs was much lower than that at a saturation concentration of 100 ppm. There was no significant difference in the extent of dechlorination between surfactant-free and -amended sediments. Moreover, surfactant did not change the congener specificity or broaden the congener spectrum for dechlorination at PCB concentrations below 40 ppm. Taken together, it seems that at a given PCB concentration, dechlorination characteristics of dechlorinating populations may be determined by not only the congener specificity of the microorganisms but also the affinity of dechlorinating enzyme(s) to individual PCB congeners.
Microbial Production and Characterization of Superparamagnetic Magnetite Nanoparticles by Shewanella sp. HN-41
Lee, Ji-Hoon ; Roh, Yul ; Hur, Hor-Gil ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1572~1577
A facultative dissimilatory metal-reducing bacterium, Shewanella sp. strain HN-41, was used to produce magnetite nanoparticles from a precursor, poorly crystalline iron-oxyhydroxide akaganeite (
-FeOOH), by reducing Fe(III). The diameter of the biogenic magnetite nanoparticles ranged from 26 nm to 38 nm, characterized by dynamic light scattering spectrophotometry. The magnetite nanoparticles consisted of mostly uniformly shaped spheres, which were identified by electron microscopy. The magnetometry revealed the superparamagnetic property of the magnetic nanoparticles. The atomic structure of the biogenic magnetite, which was determined by extended X-ray absorption fine structure spectroscopic analysis, showed similar atomic structural parameters, such as atomic distances and coordinations, to typical magnetite mineral.
Beneficial Effects of Fluorescent Pseudomonads on Seed Germination, Growth Promotion, and Suppression of Charcoal Rot in Groundnut (Arachis hypogea L.)
Shweta, Bhatia ; Maheshwari, Dinesh Kumar ; Dubey, Ramesh Chand ; Arora, Daljit Singh ; Bajpai, Vivek K. ; Kang, Sun-Chul ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1578~1583
Rhizobacteria are used as inoculants to enhance crop yield and for biological control of fungal pathogens. Fluorescent pseudomonads isolated from the rhizosphere of groundnut showed suppression of the phytopathogen Macrophomina phaseolina that causes charcoal rot of groundnut, an economically important agroproduct. Two strains of fluorescent pseudomonads, designated as PS1 and PS2, were selected as a result of in vitro antifungal activity. After 5 days of incubation at
, both PS1 and PS2 caused clear inhibition zones in dual cultures, restricting the growth of M. phaseolina by 71 % and 74%, respectively. Both the strains were capable of producing siderophores, indole acetic acid, and hydrocyanic acid, and causing phosphate solubilization under normal growth conditions. These strains, when used as inoculants in groundnut, enhanced germination up to 15% and 30% with subsequent increase in grain yield by 66% and 77%, respectively. Conversely, when the pathogen alone was tested 57% decrease in yield was recorded. Thus the studies revealed the potential of the two pseudomonads not only as biocontrol agents against M. phaseolina, but also as a good growth promoter for groundnut.
Gene Mutations of 23S rRNA Associated with Clarithromycin Resistance in Helicobacter pylori Strains Isolated from Korean Patients
Kim, Jung-Mogg ; Kim, Joo-Sung ; Kim, Na-Young ; Kim, Yeoung-Jeon ; Kim, In-Young ; Chee, Young-Joon ; Lee, Chul-Hoon ; Jung, Hyun-Chae ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1584~1589
Although resistance of Helicobacter pylori to clarithromycin is a major cause of failure of eradication therapies, little information is available regarding gene mutations of clarithromycin-resistant primary and secondary H. pylori isolates in Korea. In the present study, we examined gene mutations of H. pylori 238 rRNA responsible for resistance to clarithromycin. DNA sequences of the 238 rRNA gene in 21 primary clarithromycin-resistant and 64 secondary clarithromycin-resistant strains were determined by PCR amplification and nucleotide sequence analyses. Two mutations of the 238 rRNA gene, A2143G and T2182C, were observed in primary clarithromycin-resistant isolates. In secondary isolates, dual mutation of A2143G+T2182C was frequently observed. In addition, A2143G+T2182C+ T2190C, A2143G+T2182C+C2195T, and A2143G+T2182C+A2223G were observed in secondary isolates. Furthermore, macrolide binding was tested on purified ribosomes isolated from T2182C or A2143C mutant strains with
erythromycin. Erythromycin binding increased in a dose-dependent manner for the susceptible strain but not for the mutant strains. These results indicate that secondary isolates show a greater variety of 238 rRNA gene mutation types than primary isolates, and triple mutations of secondary isolates are associated with A2143G+T2182C in H. pylori isolated from Korean patients.
Age- and Area-Dependent Distinct Effects of Ethanol on Bax and Bcl-2 Expression in Prenatal Rat Brain
Lee, Hae-Young ; Naha, Nibedita ; Kim, Jong-Hun ; Jo, Mi-Ja ; Min, Kwan-Sik ; Seong, Hwan-Hoo ; Shin, Dong-Hoon ; Kim, Myeong-Ok ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1590~1598
Cell proliferation and differentiation are critical processes in a developing fetal rat brain, during which programmed cell death (PCD) also plays an important role. One of the decisive factors for PCD is Bcl-2 family proteins, where Bax induces cell death, whereas Bcl-2 acts as an inhibitor of PCD. As maternal drinking is known to cause fetal alcohol syndrome (FAS) or malformation of the fetal brain during pregnancy, the objective of the present study was to investigate whether maternal ethanol exposure alters the PCD-related Bax and Bcl-2 protein expression during fetal brain development. Pregnant female rats were orally treated with 10% ethanol and the subsequent expressions of the Bax and Bcl-2 proteins examined in the fetal brain, including the forebrain, midbrain, and hindbrain, from gestational day (GD) 15.5 to GD 19.5, using Western blots, in situ hybridization, and immunohistochemistry. With regard to the ratio of Bcl-2 to Bax proteins (Bcl-2/Bax), the Bax protein was dominant in the forebrain and midbrain of the control GD 15.5 fetuses, except for the hindbrain, when compared with the respective ethanol-treated groups. Moreover, Bcl-2 became dominant in the midbrain of the control GD 17.5 fetuses when compared with the ethanol-treated group, representing an alternation of the natural PCD process by ethanol. Furthermore, a differential expression of the Bcl-2 and Bax proteins was found in the differentiating and migrating zones of the cortex, hippocampus, thalamus, and cerebellum. Thus, when taken together, the present results suggest that ethanol affects PCD in the cell differentiation and migration zones of the prenatal rat brain by modulating Bax and Bcl-2 expression in an age- and area-dependent manner. Therefore, this is the first evidence that ethanol may alter FAS-associated embryonic brain development through the alteration of Bax and Bc1-2 expression.
Lymphatic Delivery of
-labeled Dextran Acetate Particles Including Cyclosporine A
Kim, Jin ; Chung, Kyong-Hwan ; Lee, Chang-Moon ; Seo, Young-Soon ; Song, Ho-Chun ; Lee, Ki-Young ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1599~1605
Biodistribution and lymphoscintigraphy of cyclosporine A (CyA) and technetium-99m (
) were studied using
-labeled dextran acetate (DxA) including CyA. DxA particles were prepared from dextran with acetic anhydride, and CyA was loaded into them. Lymphatic delivery of
-labeled DxA particles containing CyA was evaluated after subcutaneous injection into the foot pad of rats and compared with those of
-labeled human serum albumin (HSA). The labeling efficiency of CyA-loaded
-DxA particles was about 95% at 30 min. The labeling efficiency maintained stably above 80% for 12 h. The percent injected dose (%ID) of CyA-loaded
-DxA was similar to that of
-HSA at the inguinal lymph node after 40 min. The CyA-loaded
-DxA could be as well distributed as
-HSA through the lymph node. The DxA particles could steadily distribute the CyA as well as the
radiolabeling through the lymph node.
Detection of Expressed IL-32 in Human Stomach Cancer Using ELISA and Immunostaining
Seo, Eun-Hee ; Kang, Jeong-Woo ; Kim, Ki-Hong ; Cho, Min-Chul ; Lee, So-Jung ; Kim, Hee-Jong ; Kim, Jung-Hee ; Kim, Eun-Jin ; Park, Dong-Ki ; Kim, Soo-Hyun ; Choi, Yang-Kyu ; Kim, Jin-Man ; Hong, Jin-Tae ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1606~1612
Interleukin (IL)-32 is a recently identified proinflammatory cytokine that is one of the IL-18 inducible genes, and plays an important role in autoimmune and inflammatory diseases. We produced antibodies against IL-32 and studied the expression of IL-32 in human stomach cancer. We detected IL-32 secreted from K-562 cells which were stably transfected with IL-32 and in the sera of stomach cancer patients by a sandwich ELISA using a monoclonal antibody KU32-52 and a polyclonal antibody. In order to optimize a sandwich immunoassay, recombinant IL-32a was added, followed by the addition of a biotinylated KU32-52 into microtiter plate wells precoated with a goat anti-IL-32 antibody. The bound biotinylated KU32-52 was probed with a streptavidin conjugated to HRP. This sandwich ELISA was highly specific and had a minimal detection limit of 80 pg/ml (mean
SD of zero calibrator) and measuring up to 3,000 pg/ml. This ELISA showed no cross-reaction with other cytokines such as hIL-1
, hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-
. Intra-assay coefficients of variation were 18.5% to 4.6% (n=10), and inter-assay coefficients were 23% to 9% (n=10). The average IL-32 level in the sera of 16 stomach cancer patients (189 pg/ml) was higher than that of 12 healthy control men (109 pg/ml). Our results indicate that serum IL-32 level can be detected by using an established ELISA, and that this immunoassay and mAb KU32-09 specific for immunohistochemistry can be used in the detection of expressed and secreted IL-32 in stomach cancer patients.
Active Component of Fatsia japonica Enhances the Transduction Efficiency of Tat-SOD Fusion Protein both In Vitro and In Vivo
Lee, Sun-Hwa ; Kim, So-Young ; Kim, Dae-Won ; Jang, Sang-Ho ; Lim, Soon-Sung ; Kwon, Hyung-Joo ; Kang, Tae-Cheon ; Won, Moo-Ho ; Kang, Il-Jun ; Lee, Kil-Soo ; Park, Jin-Seu ; Eum, Won-Sik ; Choi, Soo-Young ;
Journal of Microbiology and Biotechnology, volume 18, issue 9, 2008, Pages 1613~1619
It has been reported that Tat-SOD can be directly transduced into mammalian cells and skin and acts as a potential therapeutic protein in various diseases. To isolate the compound that can enhance the transduction efficiency of Tat-SOD, we screened a number of natural products. 3-O-[
-L-arabinopyranosyll-hederagenin (OGAH) was identified as an active component of Fatsia japonica and is known as triterpenoid glycosides (hederagenin saponins). OGAH enhanced the transduction efficiencies of Tat-SOD into HeLa cells and mice skin. The enzymatic activities in the presence of OGAH were markedly increased in vitro and in vivo when compared with the controls. Although the mechanism is not fully understood, we suggest that OGAH, the active component of Fatsia japonica, might change the conformation of the membrane structure and it may be useful as an ingredient in anti-aging cosmetics or as a stimulator of therapeutic proteins that can be used in various disorders related to reactive oxygen species (ROS).