Background: Single nucleotide polymorphisms (SNPs), which consist of a substitution of a single nucleotide pair, are the most abundant form of genetic variations occurring with a frequency of approximately 1 per 1000 base pairs. SNPs by themselves do not cause disease but can predispose humans to disease, modify the extent or severity of the disease or influence the drug response and treatment efficacy. Single nucleotide polymorphisms (SNPs), particularly those within the regulatory regions of the genes often influence the expression levels and can modify the disease. Studies examining the associations between SNP and the disease outcome have provided valuable insight into the disease etiology and potential therapeutic intervention. Traditionally, the genotyping of SNPs has been carried out using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP), which is a low throughput technique not amenable for use in large-scale SNP studies. Recently, TaqMan real-time PCR chemistry was adapted for use in allelic discrimination assays. This study validated the accuracy and utility of real-time PCR technology for SNPs genotyping Methods: The SNPs in promoter sequence (-37 and -524) of lung cancer suppressor gene, RRM1 (ribonucleotide reductase M1 subunit) with the genomic DNA samples of 89 subjects were genotyped using both real-time PCR and PCR-RFLP. Results: The discordance rates were 2.2% (2 mismatches) in -37 and 16.3% (15 mismatches) in -524. Auto-direct sequencing of all the mismatched samples(17 cases) were in accord with the genotypes read by real-time PCR. In addition, 138 genomic DNAs were genotyped using real-time PCR in a duplicate manner (two separated assays). Ninety-eight percent of the samples showed concordance between the two assays. Conclusion: Real-time PCR allelic discrimination assays are amenable to high-throughput genotyping and overcome many of the problematic features associated with PCR-RFLP.
Kim, Deok-Joong;Song, Yong-Beom;Park, Sang-Hee;Kim, Hyoung-Sun;Lee, Hye-Yoon;Yu, Mi-Kyung;Lee, Kwang-Won
Journal of Dental Rehabilitation and Applied Science
/
v.29
no.1
/
pp.37-44
/
2013
Sodium hypochlorite and ethylene diamine tetra acetic acid are substances usually used during endodontic treatment. Several studies found that the bonding was negated with certain irrigants and some of the used irrigants have demineralizing and chealating effects, so it was advocated to omit the etching step in etch and rinse adhesive systems. The purpose of this in vitro study was to evaluate the influence of NaOCl & EDTA on the bonding strength of ethanol wet bonding. Thirty human molars were selected and mesiodistally sectioned into halves, thus providing sixty specimens. The specimens were randomly assigned to 4 groups(n=15) according to the irrigant regimen used : (1) irrigated with distilled water for 10min (control); (2) irrigated with 5.25% NaOCl(10min), flushed with 5.25% NaOCl(1min) (3) irrigated with 5.25% NaOCl, flushed with 17% EDTA (4) irrigated with 5.25% NaOCl, flushed with 17% EDTA. Each group was acid-etched with 37% phosphoric acid(except group 4) and had their dentin surfaces dehydrated with ethanol solutions : 50%, 70%, 80%, 95%, 3x100%, 30s for each application. After dehydration, a primer( 50% all bond 3 resin + 50% ethanol) was used, followed by the adhesive(ALL-BOND 3 RESIN) application. Resin composite build-ups were then prepared using an incremental technique. Specimens were sectioned into beams and submitted to a tensile load using a Micro Tensile Tester(Bisco Inc.). The data were statistically analyzed using one-way ANOVA and Tukey HSD at p<0.5 level. There was no significant difference on G1(control) and G2(irrigated with NaOCl only ). (p>0.05). G3(flushed with EDTA) showed significantly high tensile bonding strength compared to the G2 (p<0.05). G4( treated with EDTA but no acid-etching) was significantly lower value than G3. (p<0.05) Although there was no significant difference, 5.25% NaOCl seemed to have an adverse effect on the bonding strength of ethanol wet bonding. The flushing with EDTA after NaOCl irrigation prevents the decrease of bonding strength. The use of 17% EDTA as a final flush can enhance the bonding strength but EDTA flushing can't substitute for a acid-etching.
Journal of the korean academy of Pediatric Dentistry
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v.35
no.4
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pp.619-627
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2008
The objective of this study was to compare the microleakage of five adhesive systems in the enamel and dentin of permanent teeth. Class V cavity preparations with occlusal margins in enamel and gingival margins in dentin were prepared on both buccal and lingual surfaces of 25 extracted human molar teeth. The tested adhesives were: Adper Scotchbond Multi-purpose Plus Adhesive (SM), Adper Single bond 2 (SB), Clearfil SE Bond (SE), Adper Prompt L-Pop (PL) and G-Bond (GB). The results were as follows: 1. At the enamel margins, PL showed the highest leakage value(0.85), and others showed values of SB(0.55), GB(0.50), SM(0.35) and SE(0.25) in decreasing order. There were statistically significant differences in PL vs. SM and PL vs. SE(p<0.05). 2. At the dentin margins, GB showed the highest leakage value(2.10), and others showed values of SE(1.45), PL(1.40), SB(1.05), SM(0.70) in decreasing order. There were statistically significant differences in GB vs. SB and GB vs. SM(p<0.05). 3. Dentin margins showed high dye penetration rate than enamel margins in all material tested groups and there were statistically significant differences for SE, PL and GB.
Purpose: In our nuclear medicine department, we suggested AMC RBC labeling method improved by modifying a part of existing modified in-vitro method to raise the efficiency of $^{99m}Tc$-RBC labeling. However, it needs to be more additional time and efforts than existing modified in-vitro method because the AMC RBC labeling method has to carry out the centrifugal separation process for 3~5 minutes. Therefore, in this study, we conducted researches to aim to maintain stable labeling effects and supplement a problem about additional time by reducing rotating time when labeling $^{99m}Tc$-RBC. Materials and Methods: This research has been conducted the object of 30 patients who examined study using $^{99m}Tc$-RBC and agreed to this research at our hospital from May 2009 to September 2009. We made 4 blood samples which consisted of ACD 1 cc along with 5 cc blood from each patient and used the AMC RBC labeling method. At this moment, each labeling efficiency was calculated by different rotating time 5 min, 10 min, 15 min, and 20 min and then we compared differences. Results: As a result, When comparing the $^{99m}Tc$-RBC labeling method efficiency by using the AMC RBC labeling method which differents from rotating time, each labeling efficiency were $92.3{\pm}5.0%$ in 5 min, $95.9{\pm}5.0%$ in 10 min, $97.4{\pm}4.9%$ in 15 min and $97.7{\pm}4.8%$ in 20 min. We analyzed differences of the labeling efficiency from change of rotating time by using an one-way ANOVA and verified that in Duncan method. There was relatively efficiency low in 5min rotating time and no statistically significant change in over. Conclusions: When comparing a existing method, the AMC RBC labeling method which goes through the centrifugal separation process again offers more favorable condition to combine RBC with $^{99m}TcO4^-$ by eliminating an plasma ingredient. When using the modified in-vitro method, we have almost 20 min to rotate to acquire stable labeling efficiency. But, when using the AMC RBC labeling method, we acquire labeling efficiency well what we want within only 10 min to rotate. Decrease of rotating time can complement the AMC RBC labeling method which goes through the centrifugal separation process again and also provide more rapid study such as G-I bleeding study due to fast labeling.
Choi Jae-Sung;Han Woong;Kim Dong Sik;Park Jin Sik;Lee Jong Jin;Lee Dong Soo;Kim Ki-Bong
Journal of Chest Surgery
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v.38
no.5
s.250
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pp.323-334
/
2005
Background: Gene therapy is a new and promising option for the treatment of severe myocardial ischemia by therapeutic angiogenesis. The goal of this study was to elucidate the efficacy of therapeutic angiogenesis by using VEGF165 in large animals. Material and Method: Twenty-one pigs that underwent ligation of the distal left anterior descending coronary artery were randomly allocated to one of two treatments: intramyocardial injection of pCK-VEGF (VEGF) or intramyocardial injection of pCK-Null (Control). Injections were administered 30 days after ligation. Seven pigs died during the trial, but eight pigs from VEGF and six from Control survived. Echo-cardiography was performed on day 0 (preoperative) and on days 30 and 60 following coronary ligation. Gated myocardial single photon emission computed tomography imaging (SPECT) with $^{99m}Tc-labeled$ sestamibi was performed on days 30 and 60. Myocardial perfusion was assessed from the uptake of $^{99m}Tc-labeled$ sestamibi at rest. Global and regional myocardial function as well as post-infarction left ventricular remodeling were assessed from segmental wall thickening; left ventricular ejection fraction (EF); end systolic volume (ESV); and end diastolic volume (EDV) using gated SPECT and echocardiography. Myocardium of the ischemic border zone into which pCK plasmid vector had been injected was also sampled to assess micro-capillary density. Result: Micro-capillary density was significantly higher in the VEGF than in Control ($386\pm110/mm^{2}\;vs.\;291\pm127/mm^{2};\;p<0.001$). Segmental perfusion increased significantly from day 30 to day 60 after intramyocardial injection of plasmid vector in VEGF ($48.4\pm15.2\%\;vs.\;53.8\pm19.6\%;\;p<0.001$), while no significant change was observed in the Control ($45.1\pm17.0\%\;vs.\;43.4\pm17.7\%;\;p=0.186$). This resulted in a significant difference in the percentage changes between the two groups ($11.4\pm27.0\%\;increase\;vs.\;2.7\pm19.0\%\;decrease;\;p=0.003$). Segmental wall thickening increased significantly from day 30 to day 60 in both groups; the increments did not differ between groups. ESV measured using echocardiography increased significantly from day 0 to day 30 in VEGF ($22.9\pm9.9\;mL\;vs.\;32.3\pm9.1\;mL;\; p=0.006$) and in Control ($26.3\pm12.0\;mL\;vs.\;36.8\pm9.7\;mL;\;p=0.046$). EF decreased significantly in VEGF ($52.0\pm7.7\%\;vs.\;46.5\pm7.4\%;\;p=0.004$) and in Control ($48.2\pm9.2\%\;vs.\;41.6\pm10.0\%;\;p=0.028$). There was no significant change in EDV. The interval changes (days $30\~60$) of EF, ESV, and EDV did not differ significantly between groups both by gated SPECT and by echocardiography. Conclusion: Intramyocardial injection of pCK-VEGF165 induced therapeutic angiogenesis and improved myocardial perfusion. However, post-infarction remodeling and global myocardial function were not improved.
Objective: The aim of this study was to investigate the experimental brush wear pattern of a light cured surface sealant, Biscover (Bisco, Schaumburg, IL), and to evaluate its cariostatic effect. Methods: Caries-free human premolars were used for the Biscover coating group (n = 90) and the control group (n = 10). The Biscover coating group was randomly assigned to nine subgroups of 10 each and the control group was assigned to two subgroups of 5 each according to the number of brushing strokes. An experimental 3-body wear test was conducted under different strokes of wear test. Tooth-brushing was accomplished with movement of each brush head set at a frequency of 100 rpm under a force of 1.5N, Surface roughness was tested before, and after Biscover coating, and after brushing. Then, each of the 10 teeth of both groups were placed in artificial caries inducing solution for 7 days. All tooth surfaces were assessed using scanning electron microscopy. Results: Biscover coated surfaces showed a smoother texture than enamel surfaces. The roughness was increased after experimental brushing and after 10,800 brushing strokes, the whole layer of Biscover wore out. However, teeth in the Biscover coating group had a cariostatic effect in cariogenic conditions. Conclusions: We suggest that white lesions in orthodontic patients can be suppressed by topical applications of Biscover.
The mechanism by which $V_H$ region gene segments is selected in B lymphocyte is not known. Moreover, evidence for both random and nonrandom expression of $V_H$ genes in matured B cells has been presented previously. In this report, the technique of in situ hybridization allowed us to analyze expressed $V_H$ gene families in normal B lymphocyte at the single cell level. The analysis of normal B cells in this study eliminated any posssible bias resulting from transformation protocols used previously and minimized limitation associated with sampling size. Therefore, an accurate measure of the functional and expressed $V_H$ gene repertoire in B lymphocyte could be made. One of the most important controls for the optimization of in situ hybridization is to establish probe concentration and washing stringency due to the degree of nucleotide sequence similarlity between different families which in some cases can be as high as 70%. When the radioactive $C{\mu}$ and $V_{H}J558$ RNA probes are tested on LPS-stimulated adult spleen cells, $2{\sim}4{\times}106cpm$/slide shows low background and reasonable frequency of specific positive cells. For the washing condition. 40~50% formamide at $54^{\circ}C$ is found to be optimum for the $C{\mu}$. $V_{H}S107$ and $V_{H}J558$ probes. The analyzed results clearly demonstrate that the level of each different $V_H$ gene family expression is dependent upon the complexity or size of that family. These findings are also extended to the level of $V_H$ gene family expression in separated bone marrow B cells depend upon the various stage of differentiation and conclude no preferential utilization of specific $V_H$ gene family. Thus, the utilization of VH gene segments in B lymphocyte of adult BALB/c mice is random and is not regulated or changed during the differentiation of B cells.
Separating the patient from hypothermic cardiopulmonary bypass(CPB) before achieving adequate rewarming often results in afterdrop, which can predispose to electrolyte disturbances, arrhythmia, hemodynamic alterations, and shivering-induced increase of oxygen consumption. In an attempt to find an adequate end point temperature of rewarming after hypothermic CPB, 50 pediatric cardiac surgical patients were r ndomly assigned for end point temperature of rewarming of 35.5$^{\circ}C$ (Group 1) or 37t (Group 2), rectal temperature. Thereafter the rectal temperature was measured half, one, four, eight, and 16 hour after arrival to the intensive care unit(ICU), with heart rate and blood pressure. Additionally the rectal temperature was compared with esophageal temperature during CPB, and axillary temperature luring stay in the ICU. Nonpulsatile perfusion with a roller pump was used in all patients and a membrane or bubble oxygenator was used for oxygenation. Both groups were comparable with respect to age, sex, body surface area, total bypass time, and rewarming time. There was no afterdrop in both groups, and there were no statistical differences in the rectal temperatures between two groups. There were also no statistical dilyerences with respect to the heart rate and blood pressure between two groups. At the end of rewarming the esophageal temperature was higher than the rectal temperature. The axil ary temperature measured in ICU was always lower than the rectal temperature. No shivering was noted in all patients. In conclusion, with restoration of rectal temperature above 35.5$^{\circ}C$ at the end of CPB in pediatric patients, we did not observe an afterdrop.
The aims of this study were to evaluate the anti-obese effects of pine needle, black tea and green tea in rats and overweight people. Supplementation of 1, 2, 4 and 8% amount to the control diet of pine needle extract and mixture groups significantly decreased body weight gain and visceral fat mass compared with that of control diet group. Supplementation of 1, 2 and 4% amount to the control diet of black and green tea extracts groups significantly decreased body weight gain and visceral fat mass compared with that of control diet group. Anti-obese effect in body weight gain and visceral fat mass of mixture group was higher than in other groups. In human study, extracts mixture supplementation to overweight subjects significantly decreased both body weight and body fat compared with placebo control group. In vitro study, black and green tea extracts significantly inhibited both the pancreatic lipase and ${\alpha}-amylase$ activities dose dependently. In conclusion, the anti-obese effects of pine needle, black, and green teas in rats were found. In overweight human subjects, extracts mixture decreased body weight and body fat compared with placebo control group. Anti-obese effect in black and green tea groups might be from an decrease in carbohydrate and fat digestions via inhibition of pancreatic ${\alpha}-amylase$ and lipase activities in part.
Purpose : The aim of this study is to determine and compare the effects of adjunctive therapy with different doses of recombinant human granulocyte-colony stimulating factor(rhG-CSF) on reversing sepsis-associated neonatal neutropenia, and their survival rate in a group I/II-type trial. Methods : RhG-CSF was injected subcutaneously to 10 septic-neutropenic neonates with doses of $10{\mu}g/kg$ from Oct. 1995 to Sep. 1996, and was administered to another 12 septic-neutropenic neonates with doses of $5{\mu}g/kg$ from Oct. 1996 to Sep. 1997. Neutrophilic responses and the outcomes of both groups were compared. Results : In the rhG-CSF $10{\mu}g/kg$ treated group and in the $5{\mu}g/kg$ treated group, the absolute neutrophil count(ANC) was $1,065{\pm}89$($mean{\pm}SEM$) and $1,053{\pm}131$, respectively. The only difference between the two groups was the peak ANC at 48 hours. Eight patients from the remaining nine of rhG-CSF $10{\mu}g/kg$ treated group(88.9%) and ten in $5{\mu}g/kg$ treated group(83.3%) survived the sepsis and were discharged without any problems. Conclusions : RhG-CSF can increase the neutrophil count in critically ill septic neutropenic neonats. The survival rate of both groups were up to 90%. This finding suggests that both doses of rhG-CSF may be effective in a therapeutically useful time frame to treat septic neonates with neonatal neutropenia attributable to bone marrow supression or neutrophil consumption.
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