• Toxicological Research
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• v.8 no.2
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• pp.191-203
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• 1992

The immunosuppressive effects of safrole were studied in female BALB/c mouse. Mice were given 100,200and 400mg safrole/kg daily for 14days and evaluated on day 15. The day 4 immunogloblin-M antibody response to T-dependent antigen, sheep red blood cells (SRBC) was inhibited dose-dependently in all doses studied. In vitro antibody response to polyclonal antigen, lipopolysaccharide (LPS) by spleen cell suspensions from safrole-treated mice were also significantly inhibited. When safrole was treated for 14days to mice, and mitogen-induced proliferation of splenocytes were assayed on day 15, there were significant suppression of responses to B-cell mitogen, LPS and T-cell mitogen concanavalin A(Con A) at a dose of 400mg safrole/kg. Direct addition of safrole on the splenocyte culture also produced a dose dependent suppression on in vitro antibody response to LPS, and mitogen-induced lymphoproliferatin at doses of 100,200,400 and 800${\mu}M$ safrole. The role of metabolic activation in safrole-induced suppression of in vitro antibody response was studied using splenocyte-hepatocyte coculture system. The suppression of in vitro antibody respose to LPS by safrole was not altered when safrole were incubated in the splenocyte-hepatocyte system for 4hr as compared with direct addition of safrole in splenocytes culture. Neither the addition of salicylamide, sulfotransferase inhibitor, nor the addation of inorganic sulfate, sulfation cofactor to the splenocyte-hepatocyte coculture, altered the suppression of antibody response by safrole. These results suggest that the immunosuppression by safrole may not by produced by the reactive metabolites which are mediated in carcinogenesis of safrole.

• The Journal of Korean Obstetrics and Gynecology
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• v.30 no.4
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• pp.18-44
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• 2017

Purpose: The object of this study was to observe the anti-climacterium activity of Gagamguibiondam-tang (GGOT) on ovariectomized (OVX) rats, a well-documented rodent models resembles with women postmenopausal climacterium symptoms, as including cardiovascular diseases, obesity, hyperlipidemia, osteoporosis, organ steatosis and mental disorders. Methods: In this study, anti-climacteric effects were evaluated separated into three categories; 1) anti-obese, 2) anti-uterine atrophy and 3) anti-osteoporotic effects. Five groups were used (8 rats in each group); sham control, OVX control, GGOT 500, 250 and 125 mg/kg administered groups. Twenty-eight days after bilateral OVX surgery, GGOT were orally administered, once a day for 84 days, and then the changes on the body weight and gain during experimental periods, serum estradiol levels, abdominal fat pad and uterus weights with histopathology of abdominal fat pads (total thickness and mean adipocyte diameters) and uterus (total, epithelial and mucosal thickness, percentages of uterine gland regions) for anti-obese and estrogenic effects. In addition, femur, tibia and fourth or fifth lumbar vertebrae (L4 or L5) wet, dry and ash weights, mineral density (BMD), bone strength (failure load), serum osteocalcin and bone specific alkaline phosphatase (bALP) contents, histological and histomorphometrical analyses - bone mass and structure with bone resorption, were monitored for anti-osteoporosis activity. Results: As a result of OVX, noticeable increases of body weight and gains, food and water consumption, weights of abdominal fat pad deposited in dorsal abdominal cavity, serum osteocalcin levels were demonstrated in this experiment with decrease of uterus, femur, tibia and L5 weights, serum bALP and estradiol levels. In addition, marked hypertrophic changes of adipocytes located in deposited abdominal fat pads, uterine disused atrophic changes, decreases of bone mass and structures of femur, tibia and L4 were also observed in OVX control rats with dramatic increases of bone resorption markers, the Ocn and OS/BS at histopathological and histomorphometrical analysis in this study as compared with sham-operated control rats, suggesting the estrogen-deficient climacterium symptoms - obese and osteoporosis were induced by OVX, respectively. However, these estrogen-deficient climacterium symptoms induced by bilateral OVX in rats were significantly inhibited by 84 days of continuous oral treatment of GGOT 500, 250 and 125 mg/kg, respectively. Especially, GGOT 500, 250 and 125 mg/kg showed clear dose-dependent inhibitory activities on the OVX-induced climacterium signs. Conclusion: The results suggest that oral administration of GGOT 500, 250 and 125 mg/kg has clear dose-dependent favorable anti-climacterium effects - estrogenic, anti-obese and anti-osteoporotic activities in OVX rats in this experiment.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.1010-1017
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• 2007

Two alternative pathways for methionine biosynthesis are known in Corynebacterium glutamicum: one involving transsulfuration (mediated by metB and metC) and the other involving direct sulfhydrylation (mediated by metY). In this study, MetB (cystathionine ${\gamma}-synthase$) and MetY (O-acetylhomoserine sulfhydrylase) from C. glutamicum were purified to homogeneity and the biochemical parameters were compared to assess the functional and evolutionary importance of each pathway. The molecular masses of the native MetB and MetY proteins were measured to be approximately 170 and 280 kDa, respectively, showing that MetB was a homotetramer of 40-kDa subunits and MetY was a homohexamer of 45-kDa subunits. The $K_m$ values for the O-acetylhomoserine catalysis effected by MetB and MetY were 3.9 and 6.4 mM, and the maximum catalysis rates were $7.4\;(k_{cat}=21\;S^{-1})\;and\;6.0\;(k_{cat}=28\;S^{-1})\;{\mu}mol\;mg^{-1}\;min^{-1}$, respectively. This suggests that both MetB and MetY can be comparably active in vivo. Nevertheless, the $K_m$ value for sulfide ions by MetY was 8.6mM, which was too high, considering the physiological condition. Moreover, MetB was active at a broad range of temperatures $(30\;and\;65^{\circ}C)$ and pH (6.5 and 10.0), as compared with MetY, which was active in a range from 30 to $45^{\circ}C$ and at pH values from 7.0 to 8.5. In addition, MetY was inhibited by methionine, but MetB was not. These biochemical data may provide insight on the role of the parallel pathways of methionine biosynthesis in C. glutamicum with regard to cell physiology and evolution.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.284-291
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• 2003

Bacillus amyloliquefaciens K-42, which produces strongly a fibrinolytic enzyme, Was isolated from Ganjang, a traditional Korean soy sauce. The fibrinolytic enzyme was purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-50, gel chromatography on Sephadex G-100, and gel chromatography on Sephadex G-75 of the culture filtrate of Bacillus amyloliquefaciens K42. The purified enzyme showed the specific activity of 59.4 units per milligram, which was increased by 17.1 fold over the culture broth. And the molecular weight of purified fibrinolytic enzyme was confirmed to be about 45,000 Dalton by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme activity was relatively stable at pH 4.0-10.0 and the optimum pH was 8.0. The activity of the purified enzyme was increased by $Mg^{2+}$ , Cu$^{2+}$ but the enzyme was totally inhibited by $Ba^{2+}$ $Hg^{2+}$ In addition, the enzyme activity was potently inhibited by EDTA, EGTA and CDTA. It was concluded that the purified enzyme was a metalloprotease. And Km value was 2.03 mg/ml to fibrin.

• The Korean Journal of Mycology
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• v.47 no.4
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• pp.359-371
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• 2019

To optimize the expression and secretion of ferritin protein associated with ion storage in the mushroom, Pleurotus eryngii, a recombinant secretion vector, harboring the ferritin gene, was constructed using a pPEVPR1b vector under the control of the CaMV 35S promoter and signal sequence of pathogen related protein (PR1b). The ferritin gene was isolated from the T-Fer vector following digestion with EcoRI and HindIII. The gene was then introduced into the pPEVPR1b secretion vector, and it was then named pPEVPR1b-Fer. The recombinant vector was transferred into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformants were selected on MCM medium supplemented with kanamycin and its expression was confirmed by SDS-PAGE and western blotting. Expression of ferritin protein was optimized by modifying the culture conditions such as incubation time and temperature in batch and 20 L airlift type fermenter. The optimal conditions for ferritin production were achieved at 25℃ and after incubating for 8 days on MCM medium. The amount of ferritin protein was 2.4 mg/g mycelia, as measured by a quantitative protein assay. However, the signal sequence of PR1b (32 amino acids) seems to be correctly processed by peptidase and ferritin protein may be targeted in the apoplast region of mycelia, and it might not be secreted in the culture medium. The iron binding activity was confirmed by Perls' staining in a 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in P. eryngii mycelia. Mycelium powder containing ferritin was tested as a feed additive in broilers. The addition of ferritin powder stimulated the growth of young broilers and improved their feed efficiency and production index.

• Journal of Convergence for Information Technology
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• v.12 no.3
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• pp.244-251
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• 2022

This study attempted to investigate the applicability of Geranium maculatum extract as a cosmeceutical ingredient. For this, DPPH, ABTS and FRAP assays were performed to assess radical scavenging activities. To evaluate antioxidant substances, in addition, polyphenol and flavonoid concentrations were measured. Furthermore, cytotoxicity, whitening and anti-inflammatory tests were conducted, using B16F10 and RAW 264.7 cells, and the results found the followings: In the DPPH and ABTS assays, 265.8 mg ascorbic acid/g and 168.5 mg ascorbic acid/g of antioxidant capacities were found respectively. According to the FRAP assay, 1 mg Geranium maculatum extract was same with ascorbic acid 229±9 ㎍ in terms of reducing power. In polyphenol and flavonoid concentrations, 32.989±1.610 mg/g and 11.098±0.261 mg/g were observed each. The above results show that cells survived in the test concentrations more than 80 percent, confirming the low toxicity of Geranium maculatum extract. According to whitening testing, melanin synthesis was reduced depending on concentration, and at the same time, 40.62±2.07% of melanin production inhibition was found at 100 ㎍/mL. In anti-inflammatory testing, inflammation was reduced depending on concentration, and 27.86±2.82% of inhibition of inflammation was detected simultaneously, confirming the applicability of Geranium maculatum extract as a cosmeceutical ingredient.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1077-1086
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• 2001

The sprA and sprB gene encoding chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB) and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from Streptomyces griseus ATCC10137 and overexpressed in Streptomyces lividans TK24 as a heterologous host. The chymotrypsin activity of tole culture broth measured with the artificial chromogenic substrate , N-succinyl-ala-ala-pro-phe-p-nitroanilide, was 10, 14 and 14 units/mg in the transformants haboring the sprA, sprB and sprD genes, respectively. The growth of S. lividans reached the maximum cell mass after 4 days of culture, yet SGPA and SGPD production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The trypsin activity of the culture broth measured with the artificial chromogenic substrate , N-${\alpha}$-benzoyl-DL- arginine-p-nitroanilide , was 16 units/mg and SGT production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The introduction of the sprA gene into S, lividans TK24 triggered the biosynthesis of pigmented antibiotics, actinorhodin and undecylprodigiosin, and induced significant morphological changes in the colonies in Benedict, R2YE, and R1R2 media. In addition, the introduction of the sprT gene also induced morphological changes in the colony shape without affecting the antibiotic production, thereby implying that certain proteases would appear to play very important and specific roles in secondary-metabolites formation and morphological differentiation in Streptomyces.

• Journal of Veterinary Clinics
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• v.8 no.2
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• pp.127-142
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• 1991

This study was carried out to investigate fatty liver in Korean black goats. Adult female goats were divided into 3 test groups(A, B and C). Group A and B of goats each received 3 test consecutive daily doses of DL-ethionine at 75mg/kg and 150mg/kg body weight, respectively. Group C of goats was given 3 consecutive doses of the compound every 48 hours at 150mg/kg body weight. The clinical symptoms, hematological values, serum chemical values and histopathological study of the liver were investigated in the test animals. The results obtained are as follows ; 1. Fatty liver were observed in every test animal. 2. Some clinical symptoms( anorexia, depression) were appeared from 1st day to 7th day after administration of the compound in every test animal. In addition to these symptoms, diarrhea and salivation were generally observed in test animals which were given the compound at 150mg/kg body weight. The degree of these symptoms was dose dependent. 3. There was no significant variations in total WBC counts and fibrinogen values in the blood of test goats. The PCV values were significantly increased on 5th day of dosing in group A and B of goats. 4. The total lipid value was not changed but the concentration of NEFA was significantly increased on 3rd day of dosing with the compound and returned to normal value after 10 days hereafter. The value of triglycerides was significantly increased on 1st day and returned to normal value on 3rd day of dosing. The value of cholesterol was significantly decreased on 3rd day and returned to normal value on 10th day after treatment. 5. Total protein level was decreased on 10th day of dosing in the groups of B and C, and billirubin level was significantly increased on 7th day of dosing in every test group and returned to normal level after 13th day of administration. 6. The activity of GGT in serum was not changed while the activities of SDH and AST were significantly increased in every test goat and those values were returned to normal after 10~13th day of trestment. 7 The 35K-protein fraction in serum was not detected by SDS-polyacrylamide gel electrophoresis, but this protein fraction was detected by the same method after treating the 21st and 22nd fraction which were obtained by column chromatography with Sephadex G-100. 8. The affected liver was congested and swollen on 3rd day, and yellowish brown in color and mottled appearance on 7th day of treatment. Histopathologically, fat droplets were common in the hepatocytes, this change was intensive on 7th day after treatment in group B and C. Hepatic cell necrosis was observed in some livers but this pathological change was disappeared and returned to normal after 13 days of treatment.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.24 no.1
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• pp.78-85
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• 2011

Background and Objectives : The aim of this study was to investigate anti-inflammatory and anti-oxidant effects of Hyunggaeyungyo-tang(HYT) on RAW 264.7 cells. Material and Methods : Two types of experiments were implemented for this study: first, the experiment to study the anti-oxidant effect of HYT using riboflavin; second, in vitro experiment to investigate the suppression of NF-${\kappa}$B activation using RAW 264.7 cells (I${\kappa}$B kinase and induce nitric oxide synthase mRNA expression) Results : 1. The anti-oxidant effects of HYT was dose-dependantly increased. 2. The RAW 264.7cells were treated with LPS for 1 hours prior to the addition of indicated concentrations(0.4,-1.0mg/$m\ell$) of HYT, and the cells were further incubated for 24 hours. The LPS-induced IKK & iNOS mRNA expression were dose-dependantly decreased in HYT treated RAW 264.7cells. Conclusion : The results suggest that HYT is significantly effective in the treatment of inflammation through the suppression of NF-${\kappa}$B activation and iNOS production.

• The Korean Journal of Food And Nutrition
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• v.25 no.4
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• pp.807-819
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• 2012

To evaluate the functional characteristic and availability for drinking of the fermented Smilax china leaf tea by using different microbial species, various fermented leaf tea was prepared by non-fermentation (C), or the fermentation of Saccharomyces cerevisiae (S), Bacillus sp. (B), Bifidobacterium bifidus (L), Monascus pilosus (M) and Aspergilus oryzae (A), and sensory and antioxidant parameter of each brewed tea was observed. The color of the A tea was red, but the other teas were yellow in color. Furthermore, the aesthetic quality of the A and M tea was 3.95 and 3.30 point, respectively, and other teas (2.55~2.28) were similar to that of the C tea. TP of fermented tea water extract was lower than that of the C, although TF was not significantly different between the fermented and non-fermented tea. Especially, TF of the A tea was significantly lower than those of the other teas. The range of EDA ($1mg/m{\ell}$) of water and ethanol extracts of tea C and the fermented teas was 19.25~22.48%; however, tea A was only 8.04~12.49%. In addition, FRAP, FICA and LPOIA of teas were not significantly different between the fermented and non-fermented teas. On the other hand, XOIA and AOIA of tea ethanol extracts were slightly higher than those of water extracts. XOIA of water extract derived from the teas was 4.83~9.20%, while ethanol extract of these was 9.00~19.00%. However, XOIA of B and L teas water extract was not detected. Furthermore, AOIA of fermented tea water extract (30.17~48.52%) were lower than those of ethanol extract (44.09~66.93%). In this study, interestingly, antioxidant parameters, such as FRAP, FICA, LPOIA and AOIA, of the A tea water extract (0.1%) was higher than that of the other tea in spite of high decreasing rate in the contents of TP and TF. Therefore, above results imply the possibility of fermented Smilax china leaf tea as a functional food.