A new lectin was partially purified from starfish,Asterina pectinifera by means of physiological saline extraction, salt fractionation, ion exchange chromatography and hy droxyapatite chromatography, and it was named APL. The biochemical properties of the APL were characterized. In addition, its effects on lymphocyte mitogenicity and cancer cell agglutinability were tested. The APL agglutinated nonspecifically human erythrocytes and rabbit blood cells. Agglutinability was decreased to 30% of control activity below pH 5 and above pH 9 and was relatively unstable at increasing temperatures above 60$^{\circ}C$. The activity was reduced by addition of two kinds of metal ions, $Ba^{2+},\;Mn^{2+}$ and chelating agent, EDTA. APL was proved to be glycoproteins containing 9% sugars. For carbohydrate specificity, it was found that the activity of APL was inhibited by D(+)-glucosamine, D(+)-galactosamine, stachyose, N-acetyl-galactosamine and methyl-${\alpha}$-D-galactopyranoside among 35 sugars tested. In amino acid composition, the contents of acidic amino acids such as aspartic acid and glutamic acid were relatively high. This result suggest that the isoelectric point would be in a lower range. APL was found that it promotes the division of human lymphocytes. APL was proved to be a potent agglutinin for cancer cells such as HeLa, L929 and L1210 cells. Significant changes on the HeLa cell surfaces affected by APL were observed under the electron microscope.
Objective: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst.. Methods: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-fteezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of bias to cysts in experimental groups. Results: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower ($X^2$-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%,48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (${\pm}SD$) cell number of blastocyst in the exposure only ($89.2{\pm}11.5$), EM grids ($85.0{\pm}10.3$) and cryoloop ($89.0{\pm}11.0$) groups, except slow-freezing group ($79.0{\pm}10.0$), were not significantly different from that of control group ($93.1{\pm}13.9$) 24 h after thawing (Student's t-test). Conclusion: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.
Purpose: We Investigated to achieve high resolution magnetic resonance (MR) Imaging and spectra of human skin in vitro with using a 14.1 T MRI/MRS system, and to evaluate the hydration effect of a moisturizer by measuring the skin's water concentration. Materials and Methods: We used the Brukrer 14.1 T MRI/MRS system with a vertical standard bore that was equipped with a DMX spectrometer gradient system (200 G/cm at a maximum 40 A), RF resonators (2, 5 and 10 mm) and Para Vision software. Spin echo and fast spin echo pulse sequences were employed for obtaining the high resolution MR images. The 3D-localized point resolved spectroscopy (PRESS) method was used to acquire the MR spectra. Results: The high resolution MR images and spectra of human skin in vitro were successfully obtained on a 14.IT system. The water concentration of human skin after applying a moisturizer was higher than that before applying a moisturizer. Conclusions: The present study demonstrated that the high-resolution MR images and spectra of human skin from a high field MRS instrument could be applicable to evaluating the hydration state of the stratum corneum.
The one of the most important factors is the platoon design on developing AH3(Advanced Highway System), as it is related to traffic efficiency and drivers' safety. This study was evaluated that how much speed is comfortable for drivers and how long distance is appropriate for vehicular gap of platoon by measuring drivers' physiology signal and sensibility. A fixed-based AHS simulator was developed by using a real vehicle cockpit and the restructured part of Korean highway for human factors evaluation. The EEG(electroencephalogram), ECG (electrocardiogram) and GSR(Galvanic Skin Response) were measured for obtaining drivers' physiology signal according to the change of speed and gap. The brain wave(${\alpha},\;{\beta},\;{\delta},\;{\theta}$) by EEG, the response of the autonomic nervous system. the sympathetic and parasympathetic nervous system, by ECG, and relax-arousal situation by GSR were analyzed. The SD(Semantic Differential) method was also applied to evaluate drivers' sensibility by 5-grade evaluation scale with 96 adjectives. SSQ(Simulator Sickness Questionnaire) was used to measure the simulator sickness of pre and post driving, two times. As the results, drivers were comfortable with 120km/h speed of platoon and lam to 15m vehicular distance. The results of this study may differ from the adaption of the reality because of many parameters. However, the purpose of this study is show to significant results of the drivers' safety and the acceptability of human factors evaluation.
Human ribosomal protein S3 (rpS3), which has a DNA repair endonuclease activity, is a multifunctional protein. This protein is involved in DNA repair, translation, and apoptosis. In particular, rpS3 has a lyase activity, which cleaves the phosphodiester bond of damaged sites such as cyclobutane pyrimidine dimers and AP sites. Here, using deletion analysis, we identified that the repair endonuclease domain resides in the C-terminal region (165-243 aa) of rpS3. We also found that ectopic expression of GST-rpS3 in bacterial strain BL21 promoted the resistance of these cells to ultraviolet (UV) radiation and hydrogen peroxide ($H_{2}O_{2}$) treatment. The repair domain of rpS3 was sufficient to exhibit the resistance to UV irradiation and recover cell growth and viability, showing that the repair activity of rpS3 is responsible for the resistance to UV irradiation. Our study suggests that rpS3 is able to process DNA damage in bacteria via its repair domain, showing the resistance to genotoxic stress. This implies that rpS3-like activity could be operative in bacteria.
Kim, Min Jeong;Lee, Hye Hyeon;Seo, Min Jeong;Kang, Byoung Won;Park, Jeong Uck;Kim, Kyoung-Sook;Kim, Gi-Young;Joo, Woo Hong;Choi, Yung Hyun;Cho, Young-Su;Jeong, Yong Kee
Journal of Microbiology and Biotechnology
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v.22
no.12
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pp.1665-1672
/
2012
An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.
Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.
Objectives : In order to identify the awareness of influenza A (H1N1) having currently high frequency and risk as an infectious disease, to find problems and to reflect them on curriculum improvements from students before/after clinical practices. Methods : The data was collected from 279 dental hygiene students of 1st and 2nd years at G health college university from December 6th 2010 to December 10th 2010. The questionnaire were consisted of awareness of influenza A (H1N1), preventive attitude, sociodemographic characteristics. Results : 1. 1st year was 51.6%, the case having dental experience was 51.2%, in the infection control training experience 'had' was 46.6%. In the route acquiring the information, the mass media was 70.6%, in obtained information, personal hygiene was 82.1%. In the impact on human body, 'great impact' was 58.1%. In terms of the most need for response and preparedness, vaccination was the highest, 67.4%. People who experienced influenza A (H1N1) were 10.7%. 2. Awareness of influenza A (H1N1) was 0.71 points, and treatment and spreading mechanism was 0.78 points, prevention was 0.63 points, causes and definition was 0.53 points. 3. In the attitude for infection prevention of influenza A (H1N1), 'washing hands before practice' was the highest, 0.99 points and 'wear the mask only in case of contact with patient within 1~2 meters upon occurrence of no aerosol' was the lowest, 0.72 points. 4. Awareness of influenza A (H1N1) according to sociodemographic characteristics showed the significant differences upon the impacts on systemic health (p<0.05). Preventive attitude didn't show a significant difference in grade, clinical experience, experience in infection control training, acquiring rmation routes, the possibility for occurrence, impact on systemic health, the most need for prevention, experience in influenza A (H1N1) (p>0.05). 5. The significantly correlated between awareness of influenza A (H1N1) and preventive attitude(p<0.01). Conclusions : Information and preventive attitude for influenza A (H1N1) as well as systematic training programs to identify actual affecting factors and to improve the practice are needed. Also government's institutional support is needed.
Kim, Ha-Won;Shim, Mi-Ja;Choi, Eung-Chil;Kim, Byoung-Kak
Archives of Pharmacal Research
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v.20
no.5
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pp.425-431
/
1997
To examine components of Ganoderma lucidum for anti-human immunodeficiency virus (HIV) activity, the aqueous extracts of its basidiocarps were separated into high-molecular-weight (HMF) and low-molecular-weight (LMF) fractions. These fractions were used in XTT [2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide] antiviral assay which can quantitatively measure cytopathic effects of HIV-1 on CEM, human T lymphoblastoid cell line. The CEM cell line added with serial diluted HMF or LMF was cultured in the absence or presence of HIV-1. The results showed that the LMF of the aqueous extract strongly inhibited cytopathic effect of the target cell induced by HIV-1. When two-fold serially diluted LMF ranging from $40.97{\mu}g/ml$4 to 125.00 .mu.g/ml was added to the virus-free culture system, no toxicity on the target cells was detected in all the concentrations tested. However, when it was added to the HIV-infected culture system, the viabilities of the target cell reached a plateau recovering its viabilities to 71.7% and 82.5% in experiment-1 and -2 at 15.60 .mu.g/ml, respectively. The cell viabilities were then gradually decreased but maintained at more than 50% above 31.20 .mu.g/ml concentration. On the contrary, HMF did not prevent any HIV-induced cytopathic effect at any concentrations tested on this cell line. From these results, negligible toxicities were observed by both HMF and LMF of G. luciolum, and recovery of cell viability in HIV infected target cell was induced only by LMF of the carpophores.
The construction of athlete's tight-fitting garments by reducing the elastic knit pattern is an interesting subject, which directly influences the performance of the wearer. Therefore, relationships between the reduction rates of the men's pants pattern obtained using measurements of Size Korea 2004 and clothing fitting were explored to improve clothing comfort. Two pattern making methods were developed and the reduction rates were applied to those pants depending on the parts of human body. The four male subjects were university students in the ages of 20 to 24. Subjective wear sensations of the experimental garments were rated using seven Likert scales during 4 consecutive days. While wearing the garments, subjects were kept four various postures including waist flexion and sitting etc by requests of the researchers. Likert type scale was used for the evaluation and 7 point indicates that it gave the best fit as tight-fitting pants. As results, differences in the length and girth between two piece pants pattern(style 1) and one piece pants pattern(style 2) were 0.0$\sim$0.3cm. Between two pattern making methods, one piece pants pattern(the style 2) was superior to two piece pants pattern in terms of subjective sensation and fit. Among eight tight-fitting pants, C, D, G, H pants were superior in terms of fitting and stability of waist and hip part to the others. ill case of tight-fitting pants, the reduction rate of the pattern in the course direction, the reduction rate of T2/3(66%) was better than that of T/2(50%), but the reduction rate of the pattern in the wale direction, that of the part of crotch, which was very sensitive part, should be kept the same as suggested earlier by Ziegert(1988).
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