The major function of adipocytes is to store fat in the form of triglycerides. One of the signaling pathways known to affect adipogenesis, i.e. fat formation, is the WNT/${\beta}$-catenin pathway which inhibits the expression and activity of key regulators of adipogenesis. The purpose of this research is to find genes among the WNT/${\beta}$-catenin pathway which regulate adipogenesis by using small interfering (si) RNA and to find the association of single nucleotide polymorphisms (SNPs) of the gene with serum triglyceride levels in the human population. To elucidate the effects of ${\beta}$-catenin siRNA on adipogenesis key factors, PPAR${\gamma}$ and C/EBP${\alpha}$, we performed real-time PCR and western blotting experiments for the analyses of mRNA and protein levels. It was found that the siRNA-mediated knockdown of ${\beta}$-catenin upregulates adipogenesis key factors. However, upstream regulators of the WNT/${\beta}$-catenin pathway, such as DVL2 and LRP6, had no significant effects compared to ${\beta}$-catenin. These results indicate that ${\beta}$-catenin is a candidate gene for human fat accumulation. In general, serum triglyceride level is a good indicator of fat accumulation in humans. According to statistical analyses of the association between serum triglyceride level and SNPs of ${\beta}$-catenin, -10,288 C>T SNP (rs7630377) in the promoter region was significantly associated with serum triglyceride levels (p<0.05) in 290 Korean subjects. On the other hand, serum cholesterol levels were not significantly associated with SNPs of the ${\beta}$-catenin gene. The results of this study showed that ${\beta}$-catenin is associated with fat accumulation both in vitro and in the human population.
Annex I parties continued its consideration of how to address, the definitions, modalities, rules and guidelines for the treatment of Land Use, Land-use Change and Forestry (LULUCF) in the second commitment period of the Kyoto Protocol by the year of 2009. In the AWG-KP conference held in Accra, Ghana in 2008, four alternatives (gross-net carbon accounting, net-net with base year or base period accounting, net-net with forward looking baseline accounting, and land-based accounting method) for negotiations were decided in order to revise gross-net accounting method applied during the first commitment period of the Kyoto Protocol. In this study, alternative scenarios are set in consideration with reporting system (voluntary or compulsory), discount factors and cap about these three alternatives except for the method of net-net with forward looking baseline accounting, and then estimates the Removal Unit (RMU) among the countries. In the case that article 3.4 activities under the Kyoto Protocol revises from voluntary reporting to mandatory reporting, it is estimated that the loss of RMU would be huge in Russia, Australia, New Zealand, as well as Canada potentially. Net-net with base year or base period carbon accounting and land-based carbon accounting method have big difference of RMU in accordance with the base year or the base period. So the more unfavorable the country with a lot of old-age forests was, the closer the base year or period comes to the commitment period in the context of RMU. If it is getting lowered for the current rate of 85% in discount factors, RMU is getting higher to the whole countries. Therefore in Korea with little potential for afforestation and reforestation, there was the most sensitive response to the change of discount factors. Post-2012 LULUCF hereafter, it is strongly expected for the succession of current carbon accounting system which is voluntary reporting of gross-net carbon accounting and the activity for article 3.4. Other carbon accounting method is hard to accept in aspect that there is big differentiated interests among the countries and it is required enormous cost and time to develop reliable method. Provide for Post-2012 mandatory greenhouse gas reduction, Korea needs to have a competitive negotiation strategies differentiated from Annex I countries. The most reliable alternative would be to lower the discounting factors about the activities for forest management.
$^{99m}Tc-galactosyl$ human serum albumin (Tc-GSA) is a radiopharmaceutical that binds to asialoglycoprotein receptors, which are specifically present in the hepatocyte membrane. Because these receptors are decreased in hepatic parenchymal damage, the degree of Tc-GSA accumulation in the liver correlates with findings of liver function test. Hepatic images were performed with Tc-GSA in patients with acute hepatic dysfunction by Amantia Subjunquillea poisoning, and compared with these of liver ultrasonography (USG). Tc-GSA (185 MBq, 3 mg of GSA) was injected intravenously, and dynamic images were recorded for 30 minutes. Time-activity curves for the heart and liver were generated from regions of interest for the whole liver and precordium. Degree of hepatic uptake and clearance rate of Tc-GSA were generated by visual interpretation and semiquantitative analysis parameters (receptor index : LHL15 and index of blood clearance : HH15). Visual assessment of GSA scintigraphy revealed mildly decreased liver uptake in all of subjects. The mean LHL15 and HH15 were 0.886 and 0.621, graded as mild dysfunction in 2, and mild to moderate dysfunction in 1 subject. In contrast, liver USG showed no remarkable changes of hepatic parenchyme. Tc-GSA scintigraphy was considered as a useful imaging modality in the assessment of the hepatic dysfunction.
Among the nuclear medicine imaging methods available today, $H_2^{15}O-PET$ is most widely used by cognitive neuroscientists to examine regional brain function via the measurement of regional cerebral blood flow (rCBF). The short half-life of the radioactively labeled probe, $^{15}O$, often allows repeated measures from the same subjects in many different task conditions. $H_2^{15}O-$ PET, however, has technical limitations relative to other methods of functional neuroimaging, e.g., fMRI, including relatively poor time and spatial resolutions, and, frequently, insufficient statistical power for analysis of individual subjects. However, recent technical developments, such as the 3-D acquisition method provide relatively good image quality with a smaller radioactive dosage, which in turn results in more PET scans from each individual, thus providing sufficient statistical power for the analysis of individual subject's data. Furthermore, the noise free scanner environment $H_2^{15}O$ PET, along with discrete acquisition of data for each task condition, are important advantages of PET over other functional imaging methods regarding studying state-dependent changes in brain activity. This review presents both the limitations and advantages of $^{15}O-PET$, and outlines the design of efficient PET protocols, using examples of recent PET studies both in the normal healthy population, and in the clinical population.
The femoral nerve innervates the quadriceps muscles and its dermatome supplies anteromedial thigh and medial foot. Paralysis of the quadriceps muscles due to the injury of the femoral nerve results in disability of the knee joint extension and loss of sensory of the thigh. A child could walk independently even though he had injured his femoral nerve severely due to the penetrating wound in the medial thigh. We measured and analyzed his gait performance in order to find the mechanisms that enabled him to walk independently. The child was eleven-year-old boy and he could not extend his knee voluntarily at all during a month after the injury. His gait analysis was performed five times (GA1~GA5) for sixteen months. His temporal-spatial parameters were not significantly different after the GA2 or GA3 test, and significant asymmetry was not observed except the single support time in GA1 results. The Lower limb joint angles in affected side had large differences in GA1 compared with the normal normative patterns. There were little knee joint flexion and extension motion during the stance phase in GA1 The maximum ankle plantar/dorsi flexion angles and the maximum knee extension angles were different from the normal values in the sound side. Asymmetries of the joint angles were analyzed by using the peak values. Significant asymmetries were found in GA1with seven parameters (ankle: peak planter flexion angle in stance phase, range of motion; ROM, knee: peak flexion angles during both stance and swing phase, ROM, hip: peak extension angle, ROM) while only two parameters (maximum hip extension angle and ROM of hip joint) had significant differences in GA5. The mid-stance valleys were not observed in both right and left sides of vertical ground reaction force (GRF) in the GA1, GA2. The loading response peak was far larger than the terminal stance peak of vertical ground reaction curve in the affected side of the GA3, GA4, GA5. The measured joint moment curves of the GA1, GA2, GA3 had large deviations and all of kinetic results had differences with the normal patterns. EMG signals described an absence of the rectus femoris muscle activity in the GA1 and GA2 (affected side). The EMG signals were detected in the GA3 and GA4 but their patterns were not normal yet, then their normal patterns were detected in the GA5. Through these following gait analysis of a child who had selective injuries on the knee extensor muscles, we could verify the actual functions of the knee extensor muscles during gait, and we also could observe his recovery and asymmetry with quantitative data during his rehabilitation.
Journal of the Korea institute for structural maintenance and inspection
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v.24
no.3
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pp.47-56
/
2020
In this study, three levels of W/B(Water to Binder) ratio (0.37, 0.42, 0.47) and substitution ratio of GGBFS (Ground Granulated Blast Furnace Slag) rate (0 %, 30 %, 50 %) were considered to perform RCPT (Rapid Chloride Diffusion Test) at the 1,095 aged day. Accelerated chloride diffusion coefficient and passed charge of each concrete mixture were assessed according to Tang's method and ASTM C 1202, and improving behaviors of durability performance with increasing aged days are analyzed based on the test results of previous aged days from the preceding study. As the age of concrete increases, the passed charge and diffusion coefficient have been significantly reduced, and especially the concrete specimens containing GGBFS showed a significantly more reduction than OPC(Ordinary Portland Cement) concrete specimen by latent hydraulic activity. In the case of OPC concrete's results of passed charge, at the 1,095 days, two of them were still in the "Moderate" class. So, if only OPC is used as the binder of concrete, the resistance performance for chloride attack is weak. In this study, the time-parameters (m) were derived based on the results of the accelerated chloride diffusion coefficient, and the deterministic and probabilistic analysis for service life were performed by assuming the design variable as a probability function. For probabilistic service life analysis, durability failure probabilities were calculated using Monte Carlo Simulation (MCS) to evaluate service life. The service life of probabilistic method were lower than that of deterministic method, since the target value of PDF (Probability of Durability Failure) was set very low at 10 %. If the target value of PDF suitable for the purpose of using structure can be set and proper variability can be considered for each design variable, it is believed that more economical durability design can be made.
This study was conducted to observe 1) the changes of cellular association in seminiferous tubles from 2 to 8 weeks of age, and 2) the cycle phenomena of seminiferous epithelia at 14 weeks of age in Japanese quail. Total 80 birds were examined at a week interval from 2 to 8 weeks, and 14 weeks of age. The results were summarized as follows: 1) The body and testis weights showed most prominent increase during 4 to 5 weeks and 6 to 8 weeks of age respectively. And also the diameters of seminiferous tubles were abruptly enlaged during 6 to 8 weeks of age. 2) Genocytes in the seminiferous tubles were still in existence at 3 weeks of age, however they did not come out after 4 weeks of age. Spermatogonia, primary spermatocytes and spermatids made their first arpearances in the seminiferous from 3, 4 and 6 weeks of age, respectively. Spermatozoa were observed for the first time at 7 weeks of age, but full spermatogenic activity was completed from 8 weeks of age. 3) At 14 weeks of age, the average weight at testis was 3.7g and its ratio to the body weight was approximately 3.0 percent. And at this age, average diameter of seminiferous tubules was 192.08 $\mu\textrm{m}$, and average numbers of spermatogonia, spermatocytes, spermatids and spermatozoa within the cross section of seminiferous tubules were 7.74, 40.81, 28.42, 104.55 and 105.98, respectively. Spermatogonia and spermatid were classfied into 2 and 3 types, respectively. 4) At 14 weeks of age, the cycle of seminiferous epithelium could be divided into S stages with following characteristics. (1) Stage I: Seminiferous tubules showing type I and II spermatids. (2) Stage II: Seminiferous tubules showing type III spermatids only. (3) Stage III: Immature spermatozoa gathered near the sertoli cytoplasm. (4) Stage IV: Forming a bundle of 15-20 spematozoa. (5) Stage V: Spermatozoa bundle leaving the sertoli cytoplasm into lumen of the seminferous tubule. 5) Usually 2-3 stages of the seminiferous epithelium cycle were concurrently appeared within a tubular cross section, and frequency of each stage from I to V within cross section of seminiferous tubules were 11.91%, 27.03%, 27.96%, 19.04% and 17.98%, respectively.
Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.
Lee, Beom Jun;Lee, Sook Jin;Kim, Tae Myoung;Kim, Dae Joong;Nam, Sang Yoon;Hyun, Sang Hwan;Kang, Jong Koo;Hong, Jin Tae;Kim, Cheul Kyu;Yun, Young Won
Korean Journal of Veterinary Research
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v.44
no.4
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pp.507-513
/
2004
Aflatoxins are produced mainly by Aspergillus flavus and Aspergillus parasiticus that grow in improperly stored cereals. Aflatoxin B1 ($AFB_1$) is a potent hepatocarcinogen in a variety of experimental animals including human beings. In spite of a high attention to the hepatocarcinogenecity of $AFB_1$, the relative toxicity of aflatoxins ($AFB_2$ and $AFG_1$) is not fully clarified. Sprague-Dawley male rats were orally administered with $AFB_1$, $AFB_2$, and $AFG_1$ at the dose of 250 ${\mu}g/kg$ (additionally including a dose of $1250{\mu}g/kg $ for $AFB_1$) body weight. Animals were then killed at 12, 24 or 48 hrs following aflatoxin exposure. Subsequently the immunohistochemical examination of p53, cytochrome p450 1A1 (CYP450 1A1), and glutathione-S-transferase placental form (GST-P) were performed. The level of the 8-OxodG in the liver was determined. Expressions of CYP450 1A1 and p53 were high in the liver of rats through 48 hrs after treatment of $AFB_1$ at the single dose of $250{\mu}g/kg $. This pattern was more clear as increasing doses. The treatment of $AFB_2$ and $AFG_1$ did not affect the expression of CYP450 1A1 but it caused weak expression of p53. The activity of GST were not found in the liver of rats treated with aflatoxins. The formation of 8-OxodG by $AFB_1$ increased in a dose-dependent manner up to 24 hrs after a single treatment of $AFB_1$ thereafter decreased to the level of control. The treatment of $AFB_2$ and $AFG_1$ did not affect the levels of 8-OxodG in the liver of rats with increasing time. These results in the present study indicate that $AFB_1$ among aflatoxins with low comparable levels is the most toxic as determined by early biomarkers such as CYP450 1A1, p53, GST-P, and 8-OxodG.
The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.
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