• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.976-983
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• 2003

Seven Escherichia coli cells defective with aerobic growth were isolated by the insertion of ${\lambda}placMu53$, a hybrid bacteriophage of ${\lambda}$ and Mu, which created a transcriptional fusion to lacZY. These insertion mutant cells were tested on an XG ($5-bromo-4-chloro-3-indolyl-{\beta}-D-galactopyranoside$) medium for anaerobic expression of lacZ by fusion to a promoter. The chromosomal DNA from these strains were digested by EcoRI, and the EcoRI fragments that contained the fused gene and lacZ sequence were identified by Southern hybridization, using lacZ containing plasmid as a probe. The EcoRI fragment from each strain was cloned and sequenced. The sequence data were compared with the GenBank database. The mutated gene of three strains, CYT4, CYT5, and OS11, was found to be identical, and it was nrdAB that encoded ribonucleoside diphosphate reductase. The gene nrdAB was at min 50.5 on the Escherichia coli linkage map and 2,348,084 on the physical map, and is involved in hemAe-related reduction-oxidation reaction. OS6 and OS14 mutant strains had insertion at min 8.3 and the mutated gene was hemB. The hemB encodes 5-aminolevulinate dehydratase or porphobilinogen synthase. The OS3 mutant had insertion in cydB at min 16.6. The cydAB encodes cytochrome d oxidase. In the case of OS1, the fusion was made with sucA, the E1 component of ${\alpha}-ketoglutarate$ dehydrogenase.

• Animal Systematics, Evolution and Diversity
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• v.18 no.1
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• pp.75-84
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• 2002

MtDNA cyt b gene was used to investigate the geographic variation of 11 populations (106 individuals) of the planktonic developing, periwinkle Littorina brevicula, throughout Eastern, Western, and Southern coastal regions in Korea. The sequence of 500 base pairs and 13 different haplotypes were determined. Haplotype LbA was predominated through the populations studied with frequence of 0.877. Haplotypes were shown different frequencies in each coastal region (0.82, 0.90, and 1.00, respectively). enetic analysis of the 61 individuals of L. brevicula from the polluted and unpolluted sites yielded 8 distinct haplotypes. Haplotype LbA also was most common, and it was shared by 0.872 of frequency among specimens.

• Korean Journal of Veterinary Research
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• v.42 no.4
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• pp.451-457
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• 2002

The purpose of the present experiment was to investigate the micronuclei (MN) frequency in cytokinesis-blocked (CB) cells after various doses of gamma-rays in two species (fish and fowl) and so to contribute to the clarification of the question whether these species are suitable as a target organism in the test system. The frequencies of binucleated cells, and gamma-ray-induced MN in CB cells at several doses were measured in three donors of two species. No binucleated cell was noted in erythrocyte. The peaks of binucleated lymphocyte formation were found at a concentration of 2% phytohaemagglutinin (PHA) and $3{\mu}g/m{\ell}$ cytochalasin B (Cyt-B) in fish at 144 hours after incubation and 2% PHA and $6{\mu}g/m{\ell}$ Cyt-B in fowl at 72 hours after incubation. But the micronucleus counts failed to show any evidence of radiation damage. Measurements performed after irradiation showed a dose-related decrease in the formation of binucleated cells in each of the donors studied. Results indicated that the assays were not suitable for this due to blastization inhibition (binucleation failure) after irradiation. We concluded that the use of CB cell from fish and fowl for detecting the results of mdiation exposure was highly questionable.

• Korean journal of applied entomology
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• v.37 no.1
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• pp.23-30
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• 1998

Restriction fragment length polymorphism (RFLP) of a DNA has been a useful tool for analyzing genetic variation. This research was performed to establish an RFLP analytic method on the mitochondrial DNA (mtDNA) of the beet armyworm, Spodoptera exigua (Hiibner). To do this, total size of the mtDNA was measured and polymerase chain reaction (PCR) primers were selected. Its mitochondrial genome size was ca. 16kb. From a serial PCR test of 29 primers refered to the compilation of Simon et al. (1994), 22 primers were selected to amplify its mtDNA fragments. These primers resulted in short (300-700 bp) or long (1000-2000 bp) DNA products which represented a total or partial sequence of each of CO-I, CO-11, Cyt-B, ND-1, 12s rRNA, 16s rRNA, and some tRNAs. PCR-RFLP was performed in some variable mtDNA regions with 8 kinds of 4bp recognizing restriction enzymes. Different populations from Andong, Kyungsan, and Sunchun did not show any restriction site polymorphisms but had some length variation in certain regions of mtDNA.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.13
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• pp.5253-5257
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• 2014

Formaldehyde (FA) is an economically important chemical, and has been found to cause various types of toxic damage to the body. Formaldehyde-induced toxic damage involves reactive oxygen species (ROS) that trigger subsequent toxic effects and inflammatory responses, which may increase risk of cancer. Therefore, in the present study, we aimed to investigate the possible toxic mechanism in bone marrow caused by formaldehyde. In accordance with the principle of randomization, the mice were divided into four groups of 6 mice per group. One group was exposed to ambient air and the other three groups were exposed to different concentrations of formaldehyde (20, 40, $80mg/m^3$) for 15 days in the respective inhalation chambers, 2h a day. At the end of the 15-day experimental period, all mice were killed. Bone marrow cells were obtained. Some of those were used for the determination of blood cell numbers, bone marrow karyote numbers, CFU-F, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content; others were used for the determination of mitochondrial membrane potential (MMP), cell cycle and Bcl-2, Bax, CytC protein expression. WBC and PLT numbers in median and high dose groups were obvious reduced, but there was no change on RBC numbers. There was also reduced numbers of bone marrow karyotes and CFU-F in the high dose group. SOD activity was decreased, but MDA content was increased. MMP and Bcl-2 expression were decreased with increasing formaldehyde concentration, while expression of Bax and Cyt C was increased. We also observed change in cell cycling, and found that there was S phase arrest in the high dose group. Our study suggested that a certain concentration of formaldehyde could have toxic effects on the hematopoietic system, with oxidative stress as a critical effect.

• Journal of Photoscience
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• v.4 no.2
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• pp.41-48
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• 1997

The plasma membrane and microsomes, isolated from the cells treated with hematoporphyrm derivative (HpD) for 1 and 24 h, accumulated the aggregated porphyrin. The quantity of aggregated porphyrin was same in the plasma membrane and microsomes after isolating them from cells treated with HpD for 1 h whereas the microsomes accumulated higher quantity of aggregated porphyrin when cells were treated with HpD for 24 h. Photodynamic action of aggregated porphyrin on plasma membrane and microsomes was investigated using lipid specific fluorescent probes: 1,6-diphenyl-1,3,5-hexatrine (DPH) and 1-(4-trimethylammonium), 6-diphenyl-1,3,5-hexatrine(TMA-DPH). The time dependent anisotropy of these probes in the membranes was measured and the decay of anisotropy was analyzed using wobbling in cone model. Upon irradiation both the plasma membrane and the microsomes showed an increase in the limiting anis~)tropy and order parameter and a decrease in the cone angle of the lipid probes. The increase in the limiting anisotropy was pronounced in membranes isolated from the cells treated with HpD for 24 h. Photoinduced change in the limiting anisotropy was dependent on the duration of incubation of cells with HpD before isolating the membranes. In both the membranes. the membrane core was affected more as compared to the outer leaflet. In addition to the structural changes, a decrease in Na$^+$-K$^+$-ATPase and NADPH cyt c reductase activity was also observed upon irradiation of HpD treated cells. Inhibition in NADPH cyt c reductase was more when cells were treated with HpD for 24 h, however, Na$^+$-K$^+$-ATPase activity did not depend on the duration of the treatment of cells with HpD before irradiation. Our results suggest that the extent of photoinduced perturbations in the membranes varies as a function of duration of the treatment of cells with HpD and the membrane core is more susceptible to the photodynamic action of aggregated porphyrin.

• Bulletin of the Korean Chemical Society
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• v.34 no.4
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• pp.1232-1236
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• 2013

The Cu cation binding sites of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex have been investigated to explain the $[Cu{\cdot}DNA]$ biological activity caused by the Cu association to DNA. The structure of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex was investigated by electrospray ionization mass spectrometry (ESI-MS). The fragmentation patterns of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex were analyzed by MS/MS spectra. In the MS/MS spectra of $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex, three fragment ions were observed with the loss of d(CpG), {d(CpG) + Cyt}, and {d(CpG) + Cyt + dR}. The Cu cation binds to d(CpG) mainly by substituting the $H^+$ of phosphate group. Simultaneously, the Cu cation prefers to bind to a guanine base rather than a cytosine base. Five possible geometries were considered in the attempt to optimize the $[Cu(I){\cdot}d(CpG){\cdot}d(CpG)-2H]^{-1}$ complex structure. The ab initio calculations were performed at B3LYP/6-31G(d) level.

• Journal of Life Science
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• v.8 no.5
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• pp.604-613
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• 1998

The last enzyme of the sesquiterpen phytoalexin capsidiol synthesis in tobacco cell, 5-epi-aristolochene hydro-xylase which convert 5-epi-aristolochene (EAS) to capsidiol, was cloned by a reverse transcription polymerase chain reaction strategy and cDNA library screening. Cloned CYP-B3 contained high probability amino acid matches to known plant cytochrome P450 sequences and open reading frame with the conserved FxxGxRxCxG heme-binding region. Transcripts of CYP-B3 were not detected in control cells, but induced in elicitor-treated cells. Furthermore, CYP-B3 transcripts were induced by fungal extracts and cellulase but not by other stimuli(chilling, heat shock and 2,4-D). Induction of CYP-B3 transcripts by elicitor treatment was not affected by ancymidol and ketoconazole treat-ments suggesting that an inhibition of hydroxylase activity by Cyt P450 inhibitors resulting from post translational processing event.

• Asian-Australasian Journal of Animal Sciences
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• v.4 no.2
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• pp.161-164
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• 1991

A rapid and accurate method is described for the determination of nucleo-bases in rumen micro-organisms. A procedure to satisfactorily hydrolyse the micro-organisms involving reaction with a mixture of readily volatile organic acids (acetic and formic acids) under high pressure, is proposed, and optimal conditions for an analytical procedure with reversed phase HPLC is described. The following nucleobases contents (mmol/kg DM) of rumen micro-organisms were found: Adenine (Ade), 82.62; Guanine (Gua), 61.34; Cytosine (Cyt), 84.61; Thymine (Thy), 35.74; Uracil (Ura), 68.62; Hypoxanthine (Hxn), 13.06; Xanthine (Xn), 8.35. Total purine-N content (g/kg N) of rumen micro-organisms were 99.60. The nucleic acid N content (g/kg N) of microbial isolates were: RNA-N, 109.9; DNA-N, 50.9.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.1
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• pp.1-8
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• 2013

Understanding the cellular and molecular mechanisms involved in the development and progression of pulmonary hypertension (PH) remains imperative if we are to successfully improve the quality of life and life span of patients with the disease. A whole plethora of mechanisms are associated with the development and progression of PH. Such complexity makes it difficult to isolate one particular pathway to target clinically. Changes in intracellular free calcium concentration, the most common intracellular second messenger, can have significant impact in defining the pathogenic mechanisms leading to its development and persistence. Signaling pathways leading to the elevation of $[Ca^{2+}]_{cyt}$ contribute to pulmonary vasoconstriction, excessive proliferation of smooth muscle cells and ultimately pulmonary vascular remodeling. This current review serves to summarize the some of the most recent advances in the regulation of calcium during pulmonary hypertension.