• Journal of Radiation Protection and Research
• /
• v.25 no.4
• /
• pp.223-232
• /
• 2000

Comparative study was performed for the assessment of DNA damage and Chromosomal aberration in human lymphocyte exposed to low dose radiation using fluorescence in situ hybridization(FISH) and single cell gel electrophoresis(SCGE). Chromosomal aberrations in human lymphocytes exposed to radiation at doses of 5, 10, 30 and 50cGy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. FISH with chromosome-specific probe has been used to be a valid and rapid method fer detection of chromosome rearrangements induced by low dose radiation. The frequencies of stable translocation per cell equivalents were 0.0116, 0.0375, 0.040f, 0.0727 and 0.0814 for 0, 5, 10, 30 and 50cGy, respectively, and those of dicentric were 0.00, 0.0125, 0.174, 0.0291 and 0.0407 respectively. Radiation induced DNA damage in human lymphocyte in a dose-dependent manner at low doses from 5cGy to 50cGy, which were analysed by single tell gel electrophoresis(SCGE). From above results, FISH seemed to be useful for radiation biodosimetry by which the frequencies of stable aberrations in human lymphocyte can be observed more easily than by conventional method and SCGE also seemed to be sensitive method f9r detecting DNA damage by low dose radiation exposure, so that those methods will improve our technique to perform meaningful biodosimetry for radiation at low doses.

• Radiation Oncology Journal
• /
• v.14 no.4
• /
• pp.265-279
• /
• 1996

Damage produced by radiation elicits a complex response in mammalian cells, including growth rate changes and the induction of a variety of genes associated with growth control and apoptosis. At doses of 10,000 cGy or greater, the exposed individual was killed in a matter of minutes to a couple of days, with symptoms consistent with pathology of the central nervous system(CNS) including degenerative changes. The nature of the damage in irradiated cells underlies the unique hazards of ionizing radiation. Radiation injury to CNS is a rare event in clinical medicine, but it is catastrophic for the patient in whom it occurs. The incidence of cerebral necrosis has been reported as high as 16% for doses greater than 6,000 cGy. In this study, the effect of radiation on brain tissue was studied in vivo. Jun and p53 genes in the rat brain were induced by whole body irradiation of rat with 600Co in doses between 1 Gy and 100 Gy and analyzed for expression of jun and p53 genes at the postirradiation time up to 6 hours. Northern analyses were done using 1.8 Kb & 0.8 Kb-pGEM-2-JUN/Eco RI/Pst I fragments, 2.0 Kb-php53B/Bam HI fragment and ,1.1 Kb-pBluescript SK--ACTIN/Eco RI fragment as the digoxigenin or [${\alpha}^{32}P$] dCTPlabeled probes for Jun, p53 and ${\beta}$-actin genes, respectively. Jun gene seemed to be expressed near the threshold levels in 1 hour after irradiation of $^{60}$Co in dose less than 1 Gy and was expressed in maximum at 1 hour after irradiation of $^{60}$Co in dose of 30 Gy. Jun was expressed increasingly with time until 5 or 6 hours after irradiation of $^{60}$Co in doses of 1 Gy and 10 Gy. After irradiation of $^{60}$Co in dose between 20 Gr and 100 Gy, the expression of Jun was however increased to peak in 2 hours and decreased thereafter. p53 gene in this study also seemed to be expressed near the threshold levels in 1 hour after irradiation of $^{60}$Co in dose less than 1 Gy and was expressed in maximum at 6 hours after irradiation of $^{60}$Co in dose of 1 Gy, p53 was expressed increasingly with time until 5 or 6 hours after irradiation of $^{60}$Co in dose between 1 Gy and 40 Gy. After irradiation of $^{60}$Co in doses of 50 Gy and 100 Gy, the expression of p53 was however increased to peak in 2 hours and decreased thereafter. The expression of Jun and p53 genes was not correlative in the brain tissue from rats. It seemed to be very important for the establishment of the optimum conditions for the animal studies relevant to the responses of genes inducible on DNA damage to ionizing radiation in mammalian cells. But there are many limitations to the animal studies such as the ununiform patterns of gene expression from the tissue because of its complex compositions. It is necessary to overcome the limitations for development of in situ Northern analysis.

• Korean Journal of Microbiology
• /
• v.44 no.3
• /
• pp.212-220
• /
• 2008

A Pn10 DNA probe was introduced as a Prevotella nigrescens ATCC $33563^T$-specific DNA probe. In that study, the specificity of the Pn10 was tested with only type or reference strains of 5 oral bacterial species. The purpose of this study is to evaluate the specificity of the Pn10 using the wild type strains of P. nigrescens and is to develop the P. nigrescens ATCC $33563^T$-specific PCR primers based on the nucleotide sequence of the Pn10. The specificity of the Pn10 DNA probe was determined by Southern blot analysis. The nucleotide sequence of Pn10 DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pn10 DNA probe were hybridized with the genomic DNAs from P. nigrescens ATCC $33563^T$ and KB6. The Pn10 homologous region, KB6-Pn10, of P. nigrescens KB6 was cloned by PCR and sequenced. The Pn10 and KB6-Pn10 DNA fragments were consisted of 1,875 bp and 1,873 bp, respectively. The percent identity of the two was 98.8% and the divergence of them was 0.6%. The two primer sets (Pn10-F-AC/ Pn10-R-AC and Pn10-F-A/ Pn10-R-A), designed base on the nucleotide sequences of Pn10 DNA probe, were specific to the P. nigrescens ATCC $33563^T$. The two PCR primer sets could detect as little as 4 pg of genomic DNA of P. nigrescens ATCC $33563^T$. These results indicate that the two PCR primer sets have proven useful for the identification of P. nigrescens ATCC $33563^T$, especially with regard to the maintenance of the strain.

• Applied Biological Chemistry
• /
• v.47 no.4
• /
• pp.390-395
• /
• 2004

In order to characterize ion channels present in tomato roots, microsomes were incorporated into an artificial lipid bilayer arranged for electrophysiological analysis. Of the five different ion channels that could be found, a channel of 450 pS conductance was found most frequently. This channel displayed subconductance states of 450, 257 and 105 pS. All subconductance states showed linear current-voltage relationships. At positive holding potentials, high frequency of transient channel openings was observed; however, at negative potentials, the open times were long and open probability high. Po was 0.83 at -40 mV. When an additional 50 mM $K^+\;or\;Na^+$ was added to the cis side of bilayer, the reversal potentials shifted in the negative direction to near -10 mV. Thus, the 450 pS cation channel selects poorly between $K^+\;and\;Na^+$. In the presence of $100\;{\mu}M$ metal ions, the channel activity was severely inhibited by $La^{3+},\;Ba^{2+},\;and\;Zn^{2+}$, and Po was decreased to 0.2 or even less. However, $Al^{3+}\;and\;Cd^{2+}$ decreased the activity by only 20%. Interestingly, each metal ion showed different kinetics of channel inhibition. While $500\;{\mu}M\;La^{3+}$ inhibited the activities of all subconductance state, 1 mM $Zn^{2+}$ inhibited all except the 105 pS state. $Cd^{2+}$ changed the gating of the channel from a long-opening state to brief transient openings even at negative holding potentials. These data represent that the metal ions may have different binding sites on the channel protein and could be useful modulators and probes to investigate structural characteristics as well as the functional roles of the 450 pS channel on the root physiology.

• Journal of Life Science
• /
• v.14 no.4
• /
• pp.650-656
• /
• 2004

Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-,bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.

• Tuberculosis and Respiratory Diseases
• /
• v.46 no.4
• /
• pp.523-532
• /
• 1999

Background: With the development of the molecular biological methods, studies of the early diagnosis of lung cancer and the detection in the preneoplastic state by using genetic probes in the high risk groups are widely investigated. In lung cancer, squamous cell carcinoma is considered to progress from the normal bronchial mucosa to the preneoplastic state, and finally to the invasive carcinoma. In this study, we investigated the expression of p53 and c-erbB2 in the normal bronchi and the cancer tissues in patients with squamous cell lung cancer to evaluate the possibility of using these immunohistochemical markers as the diagnostic and prognostic parameters of patients with squamous cell lung cancer. Method: The normal and cancerous bronchial tissues of 25 patients with squamous cell carcinoma of the lung, surgically resected from May 1995 to November 1996, were immunohistochemically stained with the monoclonal antibodies to p53(DAKO-p53) and c-erbB2(phamingen 15821A) respectively. We compared the expression status of these markers between the normal bronchial mucosa and the tumor tissue, and also investigated the relationship between the expression status of these markers in tumor tissues and the pathological stage, and the survival time. Results: The pathological stage was as follows; stage I, II were found in 5 patients respectively, stage IIIA was in 8 patients, stage IIIB was in 4 patients, and stage IV was in 3 patients. The expression rate of p53 in the squamous cell lung cancer was 48%, and it was not expressed in the normal bronchial mucosa. The expression status was increased as the pathological stage advanced(p=0.0091 by test of trend). But there were no relationship between the expression of p53 and the median survival time. C-erbB2 did not yield a significantly meaningful result. Conclusion: p53 was not found in the normal bronchial mucosa, but it was expressed in 48% of the tumor tissue. And the expression rate increased as the pathological stage advanced. So it would be helpful to apply the immunochistochemical stain with p53 in the bronchial biopsy specimen in the early diagnosis trial or staging of squamous cell lung cancer.

• Restorative Dentistry and Endodontics
• /
• v.24 no.4
• /
• pp.560-569
• /
• 1999

Blood supply rather than nerve supply implies pulp vitality. To evaluate pulp vitality clinically, electric pulp test and thermal test which are based on sensory nerve response have been used in addition to many auxiliary data such as past dental history, visual inspection, radiographic examination, percussion, palpation and transillumination test. However, reactivity of the nerves to the stimulation is not synonymous with normalcy. Therefore measurement of pulpal blood flow using a laser Doppler flowmeter became a new trial to test the pulp vitality. The purpose of the present study was to evaluate normal pulpal blood flow level of maxillary teeth in adult to provide a guideline in determining the vitality of dental pulp. Pulpal blood flow was measured in maxillary central and lateral incisors, canines, first and second premolars and first molars of seventy nine adults of 22 - 30 years old using a laser Doppler flowmeter (PeriFlux 4001, Perimed Co., Stockholm, Sweden, 780 nm infrared laser, 1mW). For directly-made splints, silicone rubber impressions were taken directly from the mouth. For indirectly-made splints, alginate impressions were taken from the mouth and stone cast were made. After making depressions on the buccal surfaces of the cast teeth to indicate the hole positions, second impressions with vinyl polysyloxane putty were taken from the cast. Holes for the laser probes were made at the putty impressions 4mm above the gingival level. Laser probe (PF416 dental probe, 1.5mm) was inserted in the prepared hole and the splint was set in the mouth. After 10 minutes of patient relaxing, pulpal blood flow was recorded for 5 minutes on each tooth. The recorded flow was saved in the computer and calculated with a software 'Perisoft' version 5.1. Pulpal blood flow was also recorded in six teeth of five individuals with no response to electric pulp test and cold test, with periapical radiolucency, or with history of root canal treatment to compare with nonvital teeth. The difference between the mean flow values of each group of teeth were analyzed using one-way ANOVA and Duncan's Multiple Range test. The results were as follows: 1. The average pulpal blood flow values of all the tested teeth of each location were between 9 - 16 Perfusion Unit. Pulpal blood flow value was highest in maxillary lateral incisors, followed by first premolars, second premolars, canines, central incisors, and then first molars (p<0.01). 2. In six anterior teeth, indirectly-made splint group showed higher pulpal blood flow values than directly-made splint group (p<0.01). In posterior teeth, however, there was no significant flow value difference between directly-made splint group and indirectly-made splint one (p>0.05). 3. Teeth with vital pulps showed higher signal values than teeth with nonvital pulps (p<0.01), and the flow photographs showed heartbeat-synchronous fluctuations and vasomotions, while those were absent in non vital tooth.

• Tuberculosis and Respiratory Diseases
• /
• v.45 no.4
• /
• pp.760-765
• /
• 1998

Background: Glucose uptake has been found to be increased in cancer cells, and FDG-PET imaging is used for diagnosis of cancer using this phenomenon. However, the exact mechanism of increased glucose uptake in cancer cells has not been clarified. Recent studies demonstrated the presence of glucose transporter(GLUT) mRNA expression in gastrointestinal cancer and head and neck cancer, and suggested that GLUT may be associated with glucose uptake in cancer cells. In lung cancer cells, glucose metabolism is also known to be increased. We evaluated GLUT mRNA expression in human lung cancer cell lines in order to find out the mechanism of increased glucose uptake in lung cancer. Method: Total RNA was isolated from 15 human lung cancer cell lines and immortalized bronchial epithelial cell line(BEAS-2B). After electrophoresis of $20{\mu}g$ total RNA, Northern blot analysis was done using GLUT1 cDNA and GLUT3 cDNA as probes. Results: Thirteen of 14 human lung cancer cell lines expressed GLUT1 mRNA and 10 of 14 human lung cancer cell lines expressed GLUT3 mRNA. Eight human lung cancer cell lines expressed both GLUT mRNAs. BEAS-2B expressed GLUT1 mRNA and did not express detectable GLUT3 mRNA. Conclusion: The increase of glucose metabolism in lung cancer may be associated with GLUT1 and GLUT3 expression.

• Journal of Korean Society of Forest Science
• /
• v.94 no.6
• /
• pp.496-503
• /
• 2005

This study was conducted to investigate the influences of sapflow flux on soil water tensions and soil moisture content at the Abies holophylla plots in Gwangneung, Gyeonggido, from September to October 2004. The Abies holophylla had been planted in 1976 and thinning and pruning were carried out in 1996 and 2004. Sapflow flux was measured by the heat pulse method, and soil water tension was measured by tensiometer at hillslope and streamside. Time domain reflectometry probes (TDR) were positioned horizontally at the depth of 10, 30 and 50 cm to measure soil moisture content. All of data were recorded every 30 minutes with the dataloggers. The sapflow flux responded sensitively to rainfall, so little sapflow was detected in rainy days. The average daily sapflow flux of sample trees was 10.16l, a maximum was 15.09l, and a minimum was 0.0l. The sapflow flux's diurnal changes showed that sapflow flux increased from 9 am and up to 0.74 l/30 min. The highest sapflow flux maintained by 3 pm and decreased almost 0.0 l/30 mm after 7 pm. The average soil water tensions were low ($-141.3cmH_2O$, $-52.9cmH_2O$ and $-134.2cmH_2O$) at hillslope and high ($-6.1cmH_2O$, $-18.0cmH_2O$ and $-3.7cmH_2O$) at streamside. When the soil moisture content decreased after rainfall, the soil water tension at hillslope responded sensitively to the sapflow flux. The soil water tension decreased as the sapflow flux increased during the day time, whereas increased during the night time when the sapflow flux was not detected. On the other hand, there was no significant relationship between soil water tension and sapflow flux at streamside. Soil moisture content at hillslope decreased continuously after rain, and showed a negative correlation to sapflow flux like a soil water tension at hillslope. As considered results above, it was confirmed that the response of soil moisture tension to sapflow flux at hillslope and streamside were different.

• Journal of Oral Medicine and Pain
• /
• v.31 no.3
• /
• pp.223-229
• /
• 2006

The objectives of this study was to investigate the amount of tooth ablation and the change of intrapulpal temperature by Er:YAG laser as it relates to pulse energy and pulse repetition rate at the identical power and, thereby, to reveal which of the two parameters strongly relates with ablation efficiency and intrapulpal temperature. Extracted healthy human molar teeth were sectioned into two pieces and each specimen was irradiated within the combination of pulse energy and pulse repetition time at the same power of 3W; $300mJy{\times}10Hz$ group, $200mJy{\times}15Hz$ group, and $150mJy{\times}20Hz$ group. Each specimen comprised ten tooth specimens. A laser beam with conjunction of a water flow rate of 1.6 ml/min was applied over enamel surfaces of the specimens during 3 seconds and the ablation amount was determined by difference in weight before and after irradiation. To investigate the temperature change in the pulp according to the above groups, another five extracted healthy human molar teeth were prepared. Each tooth was embedded into resin block and the temperature-measuring probes were kept on the irradiated and the opposite walls in the dental pulp during lasing. When the power was kept constant at 3W, ablation amount increased with pulse energy rather than pulse repetition rate (p=0.000). Although intrapulpal temperature increased with pulse repetition rate, there were no significant differences among the groups and between the irradiated and the opposite pulpal walls, except at a condition of $150y{\times}20Hz$ (p=0.033). Conclusively, it is suggested that ablation efficacy is influenced by pulse energy rather than pulse repetition rate.