Mutations in the cystathionine ${\beta}$-synthase (CBS) gene cause homocystinuria, the most frequent inherited disorder in sulfur metabolism. CBS is the unique enzyme using both heme and pyridoxal 5-phosphate (PLP) for activity. Among the reported 140 mutations, one of the most common disease-causing alterations in human CBS is G307S mutation. To investigate the pathogenic mechanism of G307S by spectroscopic methods, we engineered the full length and the truncated G247S mutation of yeast CBS that is corresponding mutation to human G307S. Yeast CBS does not contain heme and thus gives a merit to study the spectroscopic properties. The UV-visible spectra of the purified full length and the truncated G247S yeast CBSs showed the total absence of PLP in the protein. The absence of PLP in G247S mutation was also confirmed by the PLP-cyanide adduct formation experiment, which was conducted by the incubation of the purified enzyme with KCN. The adducts were detected using a circular dichroism (CD) and a spectrofluorimeter. Radio isotope activity assay of full length and truncated G247S proteins also gave no activity. Our yeast G247S mutation data suggested that G307S might make the distortion of the active site so that cofactor PLP and substrate can not fit inside the active site. Our yeast CBS study addressed the reason why the G307S mutation in human CBS makes the enzyme inactive that consequently leads to severe clinical phenotype.
The reproductive cycle of the small filefish, Rudarius ercodes was investigated based on the annual variations of gonadosomatic index(GSI) and hepatosomatic index(HSI) by electronic and photic microscophy. The specimens used were collected at the coastal area of Benden island, Sizuokagen, Japan, from September 1982 to August 1983. GSI began to increase from March, starting season of longer daylength and higher water temperature, and reached the maximum value between June and August. It began to decrease from September with the lowest value appearing between November and February without any evident variation. The annual variations of HSI were not distinct in male filefish and were negatively related to GSI in female : HSI decreased in the summer season when the ovary was getting mature and reached the maximum in the winter season when the ovary was getting retrogressive. The ovary consisted of a pair of saccular structure with numerous ovarian sacs branched toward the median cavity. Oogonia divided and proliferated along the germinal epithelium of the ovarian sac. Young oocytes with basophile cytoplasm showed several scattering nucleoli along the nuclear membrane. when the oocytes growing to about 300 ${\mu}m$, nuclear membrane to disappear with nucleus migrating toward the animal pole. The regions of protoplasm were extremely confined within the animal hemisphere in which most of cytoplasms were filled with yolk materials and oil drops. After ovulation, residual follicles and growing oocytes remaining in the ovarian sacs degenerated. But perinucleatic young oocytes without follicles formed were not degenerated, and growing continuously still in the next year. Mitochondria and endoplasmic reticula in the cytoplasm remarkably increased with oocytes maturing and yolk accumulating. Those were considered to be functionally related to the yolk accumulation. Five or six layers of possible vitellogenin, oval-shaped disc structures with high electron density, appeared in the apex of follicular processes stretching to the microvilli pits of mature oocytes. Testis consisting of a pair of lobular structures in the right and left were united in the posterior seminal vesicle, Cortex of testis was composed of several seminiferous tubules, and medulla consisting of many sperm ducts connected with tubules. Steroid hormone-secreting cells with numerous endoplasmic reticula and large mitochondria of well developed cristae were recognized in the interstitial cells of the growing testis. Axial filament of spermatozoon invaginated deeply in the central cavity of the nucleus and the head formed U-shape with acrosome severely lacking, mitochondria formed large globular paranuclei at the posterior head, and microtubular axoneme of the tail represented 9+9+2 type. The annual reproductive cycles could be divided into five successive stages : growth(March to July), maturation(May to September), Spawning(mid May to early October) and resting stages(October to February). The spawning peak occurred from June to August.
Lee, Hye Mi;Kong, Bong Ju;Kwon, Soon Sik;Kim, Kyeong Jin;Kim, Hae Soo;Jeon, So Ha;Ha, Ji Hoon;Kim, Jin-Sook;Park, Soo Nam
Journal of the Society of Cosmetic Scientists of Korea
/
v.39
no.4
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pp.337-345
/
2013
In this study, the antioxidative effects of Aronia melanocarpa fruit and leaf extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of the ethylacetate and aglycone fractions of fruit extracts were 16.29 ${\mu}g/mL$, and 12.29 ${\mu}g/mL$, respectively. The free radical scavenging activity of fruit extract was higher than that of leaf extracts. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of the ethylacetate and aglycone fractions of fruit extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay showed 2.86 ${\mu}g/mL$, and 1.80 ${\mu}g/mL$, respectively. ROS scavenging activity of the aglycone fraction of fruit extracts was similar to that of L-ascorbic acid (1.50 ${\mu}g/mL$). The ROS scavenging activity of fruit extracts was higher than that of leaf extracts. The cellular protective effects of aglycone fraction of fruit extracts (${\tau}_{50}$ = 72.3 min) on the $^1O_2$-induced cellular damage of human erythrocytes especially were increased in a concentration dependent manner (5 ~ 50 ${\mu}g/mL$). ${\tau}_{50}$ (72.3 min) of the aglycone fraction showed 1.9 times higher than (+)-${\alpha}$-tocopherol (38 min), known as lipophilic antioxidant at 10 ${\mu}g/mL$. These results incidicate that A. melanocarpa fruit extracts have higher antioxidant effects than leaf extracts and could be applicable to functional cosmetics materials for antioxidants by protecting skin exposed to solar UV radiation against ROS including $^1O_2$.
Kim, Na-Mi;Koo, Bon-Suk;Lee, Seong-Kye;Hwang, Eui-Il;So, Seung-Ho;Do, Jae-Ho
Journal of Ginseng Research
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v.31
no.2
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pp.86-92
/
2007
This study was carried out to develop health & functional food by using Korean red ginseng for prevention of skin wrinkles. Effects of Korean red ginseng on the collagen biosynthesis and inhibition of matrix metalloproteinase-I (MMP-1) activity in human dermal fibroblast were investigated. Crude saponin contents of Korean red ginseng water extract (WE), Korean red ginseng ethanol extracts (EE) and Korean Red ginseng purified extracts (PE) were 72 mg/g, 107 mg/g and 220 mg/g, respectively. We incubated human fibroblast cell with Korean red ginseng component by addition of l ${\mu}g/ml$, 5 ${\mu}g/ml$, 10 ${\mu}g/ml$. Amount of collagen biosynthesis was 1.86 ng/ml in control sample and 2.85 ng/ml, 2.05 ng/ml and 2.58 ng/ml in retinoic acid, EE and PE respectively. Furthermore, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were shown 2.01 ng/ml and 3.07 ng/ml. MMP-l activities of EE, PE, $ginsenoside-Rg_1$ and $ginsenoside-Rb_1$ were decreased to 92%, 94%, 91% and 78% respectively as compared with control. Cell proliferation were showed 84-96% in the Korean red ginseng components. The antioxidative SOD activities of the Korean red ginseng components were showed 28-69%, however it was lower than that of Vitamin C. From this results, we conclude that Korean red ginseng have a anti-wrinkle effect and $ginsenoside-Rb_1$ may be considered as a more effective component.
Mast cell is a cell that functions mainly in our body with a respect to inflammation and allergic response. Bee venom has been progressed in a study as a model related to mechanism in alleviation of pain until now, but it is being progressed in a study relevant to immunocyte in anti-inflammation or anti-allergic response. The present study is aimed to present the basis related to a future study of gene, by researching the influence of melittin and MCD Peptide, which are major ingredients in Bee venom, upon the expression of gene in the mast cell strain. In this study, it dealt with melittin and MCD Peptide respectively, in the effective concentration after passing though the experiment of cytotoxicity by using human mast cell strain. Also, with the respect in the aspect of expression in gene that changes at this time, information was obtained through the technique of analyzing microarray. Through experimental statistics, when regarding a case that global M is significant in more than 1 or -1, in melittin, all 7 genes were accelerated, and 8 inhibited. In MCDP, 7 genes were accelerated and 17 genes inhibited. The function in the body to which these genes are related, was associated with the protein binding within a cell, the activation in the function of lymphocyte, the acceptor related to macrophage antigen. In cell nucleus, substance related to GABA A receptor, protein associated with cAMP reactive element, substance related to complement system No.8 and to B-cell, protein substance related to polycystic kidney disease, substance related to inflammation, and the protein substance of influencing coagulation of blood. Through these results of analysis, it could obtain more useful materials in clarifying the mechanism of action in melittin and MCD peptide, which are in charge of mainly medical action in the abdomen. Also, it is thought that an in-depth study on the influence of main ingredients in Bee venom, the wholly honey bee venom aqua upon anti-allergic response or anti-inflammation are further required.
The Journal of the Korean bone and joint tumor society
/
v.20
no.2
/
pp.74-79
/
2014
Purpose: Fibrous dysplasia is related to the mutation of gene encoding the alpha-subunit of a signal-transducing G-protein and has variable clinical course. Operation can be performed to prevent functional disorder or structural deformity. After curettage, autologous bone graft were used to fill the defects after curettage. The aim of this study is to compare the result of autogenous cancellous bone grafting and allogenic bone grafting for fibrous dysplasia. Materials and Methods: Among the patients who visit our hospital during the period of April, 1997 to October, 2013, we selected 34 patients who diagnosed fibrous dysplasia and visited our clinic over 1 year. There were 13 males and 21 females. Average age was 26.4 (range 2 to 57) years old. Autogenous bone graft (group I) in 5 cases, Non-autogenous bone graft (group II) in 30 cases. Iliac bone is used in all cases of autogenous bone graft. There were no significant difference in age, follow-up period, preoperational laboratory finding between two groups. Radiographic image was done to evaluate the recurrence of fibrous dysplasia or secondary degeneration. Results: There were four cases in recurrence (group I: 1 case, group II: 3 cases, p=0.554). In all recurrent cases, reoperations were done using curettage and autogenous iliac bone graft. There was no re-recurrence after reoperation. One case of secondary aneurysmal bone cyst was confirmed (group II) and 1 cases of pathologic fractures had developed (group I: 0 case, group II: 1 cases, p=0.559). No malignant change occurred. Conclusion: There were no significant difference between autogenous bone graft group and non-autogenous bone graft group. Our result suggested that autogenous bone graft seems to be good method to treat fibrous dysplasia, in the case of small volume of tumor lesion or non-weight bearing portion.
An empirical model to predict initial disease occurrence and subsequent progress of Alternaria leaf spot was constructed based on the modified degree day temperature and frequency of rainfall in three years field experiments. Climatic factors were analized 10-day bases, beginning April 20 to the end of August, and were used as variables for model construction. Cumulative degree portion (CDP) that is over $10^{\circ}C$ in the daily average temperature was used as a parameter to determine the relationship between temperature and initial disease occurrence. Around one hundred and sixty of CDP was needed to initiate disease incidence. This value was considered as temperature threshhold. After reaching 160 CDP, time of initial occurrence was determined by frequency of rainfall. At least four times of rainfall were necessary to be accumulated for initial occurrence of the disease after passing temperature threshhold. Disease progress after initial incidence generally followed the pattern of frequency of rainfall accumulated in those periods. Apparent infection rate (r) in the general differential equation dx/dt=xr(1-x) for individual epidemics when x is disease proportion and t is time, was a linear function of accumulation rate of rainfall frequency (Rc) and was able to be directly estimated based on the equation r=1.06Rc-0.11($R^2=0.993$). Disease severity (x) after t time could be predicted using exponential equation $[x/(1-x)]=[x_0/(1-x)]e^{(b_0+b_1R_c)t}$ derived from the differential equation, when $x_0$ is initial disease, $b_0\;and\;b_1$ are constants. There was a significant linear relationship between disease progress and cumulative number of air-borne conidia of Alternaria mali. When the cumulative number of air-borne conidia was used as an independent variable to predict disease severity, accuracy of prediction was poor with $R^2=0.3328$.
In this study, we analyzed the nutritional ingredients of drone pupae (16th to 20th instar old) to evaluate the value of bee products and provide basic data for product diversification, and the extracts prepared using these pupae were tested for physiological activity. According to the analysis of the general ingredients of the freeze-dried powder of these bee pupae, the moisture, crude protein, crude fat, and crude ash was 1.69 ± 0.07%, 48.52 ± 0.20%, 23.41 ± 0.14%, and 4.05 ± 0.02%, respectively. Vitamin C and vitamin E were 14.92 ± 0.52 mg/100 g and 6.06 ± 0.11 mg α-TE/100 g, respectively. Regarding minerals, the highest content of K (1349.13 ± 34.57 mg/100 g) and P (1323.55 ± 43.85 mg/100 g) was observed and Ca and Fe were 55.43 ± 1.51 mg/100 g and 5.49 ± 0.19 mg/100 g, respectively. The fatty acids of the water extracted freeze-dried pupae powder accounted for approximately 59.62 of saturated fatty acids and 40.38 of unsaturated fatty acids, and high-quality fatty acids such as palmitic acid (C16:0) was 35.49 ± 0.08 and oleic acid (C18:1, n-9) was 35.91 ± 0.22 (g/100 g total fatty acids). The total amino acid content was 38.99 ± 2.63 g/100 g and the free amino acid was a total of 5129.04 mg/100 g, of which 1257.68 mg/100 g was proline and 759.12 mg/100 g glutamic acid. The DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity of the drone pupae extract showed values of 0.8 for distilled water extract, 3.2 for 50% EtOH extract, 6.4 for 70% EtOH extract, and approximately 90% for 32 ㎍/mL for 100% EtOH extract. These results suggest that the main compound contributing to the antioxidant activity is a polar compound, and it is highly likely to be a low-molecular protein or a free amino acid. In conclusion, the honey bee drone pupa is excellent as a food resource and can be utilized as a new functional material for food and functional food.
So, Seung-Ho;Lee, Seong-Kye;Hwang, Eui-Il;Koo, Bon-Suk;Han, Gyeong-Ho;Kim, Na-Mi
Journal of Ginseng Research
/
v.31
no.4
/
pp.196-202
/
2007
Skin wrinkles are associated with collagen synthesis and matrix metalloproteinase-I (MMP-1) activity. This study was carried out to select optimum ratio of 3 herbs in skin health food for anti-wrinkle. Human dermal fibroblast cell was incubated with experimental samples, which were Korean red ginseng ethanol extracts (ER), Torilis fructus water extracts (WT), Corni fructus water extracts (WC) and their mixtures (WM1, WM3). And then we determined effects on collagen biosynthesis, MMP-1 activity and SOD activity in human dermal fibroblast cell. In control group, collagen biosynthesis was amounted at 474.8 ng/ml and 533.9 ng/ml, 539.3 ng/ml, 514.1 ng/ml in ER, WT and WC respectively. Furthermore, WM3 (KTNG0345) was increased to 561.45 ng/ml. MMP-1 activity of ER, WT, WC, WM1 were determined to 31.9 ng/ml, 32.85 ng/ml, 32.0 ng/ml, 31.3 ng/ml and WM3 (KTNG0345) was decreased to 28.85 ng/ml. In addition, the experimental samples showed a antioxidative activities. From this results, we conclude that Korean red ginseng ethanol extracts, Torilis fructus water extracts, Corni fructus water extracts and their mixtures have a anti-wrinkle effect and WM3 (KTNG0345) may be regarded as an optimum composition for synergic effect producing. The standardized components of KTNG0345, ginsenoside-$Rb_1$, torilin and loganin were identified at 10.85 mg/g, 0.128 mg/g and 3.92 mg/g respectively.
Benthic macroinvertebrates were investigated in Yongdam-dam and Hapcheon-dam resions from June to October 2011, Korea. Yongdam-dam and Hapcheon-dam are geographically contiguous, but they are classified as other water system. Experiment site(YE-1, 2, 3; HE-1, 2, 3) which is thought to be affected by dam, control site(YC-1, 2; HC-1, 2) which is thought not to be affected by dam were selected. Species composition, macroinvertebrate communities, composition of the functional feeding groups, habitat orientied groups and community stability were assessed Yongdam-dam and Hapcheon-dam regions. Ten sites were selected for quantitative(Surber sampler $30cm{\times}30cm$) of benthic macroinvertebrates. As a results, a total of 6,369 individuals including 69 species, 33 families, 12 orders, 6 classes and 4 phyla were recognized in Yongdam-dam region. Also, a total of 5,728 individuals including 81 species, 44 families, 13 orders, 5 classes and 4 phyla were recognized in Hapcheon-dam region. Dominance index was 0.27~0.50(mean${\pm}$SD $0.38{\pm}0.09$), diversity index was 2.22~2.97($2.67{\pm}0.29$), evenness index was 0.63~0.76($0.72{\pm}0.06$) and richness index was 4.43~7.06($5.69{\pm}0.99$) in Yongdam-dam region. Dominance index was 0.40~0.81($0.59{\pm}0.18$), diversity index was 1.40~2.39($2.00{\pm}0.43$), evenness index was 0.38~0.68($0.56{\pm}0.13$) and richness index was 4.04~5.80($4.95{\pm}0.70$) in Hapcheon-dam region. In the functional feeding groups, filtering-collectors and gathering-collectors were the highest in the whole sites. In the habitat orientied groups, burrowers, clingers and swimmers were considerably occupied in all sites. As a result of community stability analysis, experiment sites has been identified much as species high resistance and resilience to environmental changes in Yongdam-dam. Control sites has been identified much as species low resistance and resilience to environmental changes in Hapcheon-dam. Species belonging to the Igroup is considered to be important in the river ecosystem stability of large dams downstream areas.
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