Journal of the Korea Academia-Industrial cooperation Society
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v.11
no.9
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pp.3358-3365
/
2010
This study was carried out to investigate the effect of Sorbus commixta (SC), Geranium thunbergii (GT) and their mixture (SC:GT=1:1, MIX) on inhibition of bone loss and chondral defect. To examine their activities, we measured the alkaline phosphatase (ALP) activity in human osteoblast-like MG-63 cells and performed tartrate-resistant acid phosphate (TRAP) staining in osteoclast differentiated from Raw264.7 cells. To investigate the influence on chondrocyte differentiation, we performed alcian-blue staining in chondrocyte differentiated from ATDC5 cells. All of SC, GT and MIX did not increase ALP activity in MG-63 cells. However, SC and mixture (SC:GT=1:1, MIX) significantly inhibited osteoclastic differentiation. And they also induced chondrocyte differentiation. These results suggest that SC and GT may have a potential for the treatment of bone loss and chondral defect by suppression of osteoclast differentiation and stimulation of chondrocyte differentiation. Therefore, clarification of their mechanisms and active components will be needed.
In order to treat of alkaline dye-processing wastewater, alkalophilic strains biodegrading azo dye, Acid red 1, is isolated from natural system, and optimal culture conditions are examined using response surface analysis, statistical analysis system program. 15 different species which grow in alkaline culture media are isolated from the effluent and river soil discharged from wastewater treatment plant in dye industrial complex. One strain which has the best decolorization efficiency is chosen, and named as AR-1. The result of the examination of carbon, nitrogen and phosphorus sources which have influence on growth and decolorization reveals that optimum carbon, nitrogen and phosphorus sources are 1.0% fructose, 1.0% polypeptone, 1.0% yeast extract and 0.5% $K_2HPO_4$, respectively. In order to optimize of biodegradation conditions of dye by response surface analysis, the characteristics of decolorization and cell growth according to culture temperature and time are monitered. The result shows that the one is optimum 34.77$^{\circ}C$ for 12.97 hours; the other at 34.73$^{\circ}C$ for 12.96 hours. While, optimal conditions of culture that satisfy both cell growth and decolorization are the temperatures from 32.86$^{\circ}C$ to 36.36$^{\circ}C$ and the period of 10.96 to 15.75 hours, respectively.
The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.
Objectives: Osteoporosis is considered a serious human disease. We developed an extract of mixed herbs containing root of Platycodon grandiflorum (ExMH-PGR), which is expected to be effective in preventing or treating osteoporosis. The aim of this study was to investigate the anti-osteoporotic effect of ExMH-PGR in osteoblastic MC3T3-E1 cells and osteoclastic RAW 264.7 cells. Methods: To examine the anti-osteoporotic effect of ExMH-PGR, osteoblast and osteoclast differentiation were induced and cultured with various concentrations of ExMH-PGR. Alkaline phosphatase (ALP) activity, collagen synthesis, osteocalcin production, and mineralization in MC3T3-E1 cells were analyzed. Tartrate-resistant acid phosphatase (TRAP) activity and the formation of actin ring in RAW 264.7 cells were analyzed. Results: ExMH-PGR at concentration up to $25{\mu}g/mL$ significantly increased ALP activity, collagen synthesis, osteocalcin production, and mineralization in MC3T3-E1 cells. ExMH-PGR at 50 to $200{\mu}g/mL$ significantly inhibited TRAP activity and the formation of actin ring in RAW 264.7 cells. Conclusions: These results demonstrate that ExMH-PGR stimulates osteoblastic activities and inhibits osteoclastic activities in in vitro systems, suggesting that ExMH-PGR might be considered as an anti-osteoporotic candidate for treatment of osteoporosis disease.
Yoo, Han-Seok;Chung, Kang-Hyun;Lee, Kwon-Jai;Kim, Dong-Hee;An, Jeung Hee
Microbiology and Biotechnology Letters
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v.43
no.4
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pp.322-329
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2015
The effects of water extracts of Gallus gallus var. domesticus (Yeonsan ogolgye, GD) on the activities of osteoblast differentiation and the restraint of osteoclast formation were investigated. The water extract of GD in the human osteoblast "MG-63" cell, was examined in relation to alkaline phosphatase (ALP) activity and alizarin red stains. In order to observe the effects of osteoclasts formation, we analyzed RAW 264.7 cell tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains. The ALP activity of the water extract of hen and cock flesh (3 years) were 133.8% and 129.6%, respectively. The ALP activity of flesh extracts was also higher than that of the skin extracts. Concerning the effects of age, the 3 years old flesh extracts had a higher activity than that of the one year old extracts. However the activity of the 3 years old skin extracts was lower than that of the one year old extracts. For gender conditions, the ALP activity of the hen extract was higher than that of the cock. The degree bone mineralization in the three years old hen flesh exhibited the highest rate, at 124.3%, amongst all the groups. The TRAP activity of the flesh extracts of the three years old cock revealed the lowest rate, at 31.8%, compared to the control. Our results demonstrate that the water extract of GD increases bone mineralization and osteoblast differentiation activity in MG-63 cells and enhances the inhibitory activity of bone-resorption in RAW 264.7 cells. In conclusion, the water extracts of GD seem to be effective in the prevention and treatment of bone related disorders.
Experiments were carried out to establish the effects of acids in neutralization after alkaline treatment of rice straw, with which cellulosic single cell protein can be produced by cellulose utilizing bacteria, Cellulomonas flavigena KIST 321, previously isolated by authors. Following results were obtained. 1. Rice straw as carbon source was pretreated with 10 volumes of 1 normality of NH$_4$OH or NaOH(NaOH/substrate:40%, and then washed with water or neutralized with H$_3$PO$_4$, H$_2$SO$_4$, HCl and CH$_3$COOH. Among the above mentioned methods, neutralization with H$_3$PO$_4$after alkaline treatment was proved to be the most effective on its digestibility and SCP production. Dry cell 12.28g/$\ell$ and 78% digestibility were obtained. 2. When rice straw was treated with NaOH solution, the result suggested that the productibity of cell-mass was attained on treatment of rice straw with 6% of NaOH (NaOH/substrate ratio) for 15~24hrs at room temperature. 3. When rice straw was treated with NaOH, a volume of water to substrate is adequate by two or three fold and the amount of NaOH can be economized up to 5% for the weight of rice straw. 4. The steaming of rice straw at 121$^{\circ}C$ for 30min. in alkaline treatment of rice straw gave the similiar effectiveness to that at room temperature for 15~24hrs. and accelerated the sterilization of the substrate. 5. Finally, the level of inorganic phosphate in a medium was investigated. 11.2g of dry cell was produced at the concentration of 0.2%, phosphate (phosphorous level 0.04%) in medium even though treated rice straw was neutralized with HCI instead of H$_3$PO$_4$, and 12.2g/$\ell$ at the concentration of 0.3% phosphate (phosphorous 0.04%) on neutralization with H$_2$SO$_4$.
Park, Bum-Ki;Na, Jung-Heang;Hwang-Bo, Hoon;Lee, In-Jung;Kim, Kil-Yong;Kim, Yong-Woong
Korean Journal of Soil Science and Fertilizer
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v.38
no.1
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pp.15-24
/
2005
Enterobacter intermedium oxidizes glucose to gluconic acid and sequentially converts gluconic acid to 2-ketogluconic acid (2-KGA) under aerobic condition. Shaking incubation of E. intermedium in a broth medium containing 22.5 g glucose, 8.2 g gluconic acid and 40 g rock phosphate per liter resulted in $1028mg\;L^{-1}$ soluble phosphate. The culture broth was used as phosphate bio-fertilizer (PBF) in this experiment. To evaluate PBF produced by E. intermedium on lettuce growth, five treatments (PBF1/3, PBF2/3, PBF3/3, BP, and MF) were used. In MF and BP treatments, $P_2O_5$ 5.9 kg of mineral fertilizer per 10a was added, while in PBF1/3, PBF2/3, and PBF3/3 treatments, culture broth containing one third, two third, and same amount of soluble $P_2O_5$ 5.9 kg per 10a was applied, respectively. At 20, 35, and 50 days after transplanting of lettuce, plant growth components, biomass, enzyme activities and soil chemical properties were analyzed. Dehydrogenase activity and available phosphate concentration of rhizosphere in phosphate bio fertilizer treatments (PBF1/3, PBF2/3, PBF3/3) were generally higher compared to MF and BP treatments. Soil biomass in PBF3/3 treatment was significantly higher than MF and BP treatments at early growth stage. However, there was no significant difference among all treatments with time. Plant fresh weights in PBF1/3, PBF2/3, and MF treatments were better than those in BP and PBF3/3 treatments. However, in PBF2/3 treatment the highest fresh weight was discovered where alkaline phosphatase activity was generally higher than other treatments at 35 and 50 days. Enhancement of lettuce growth at 35 and 50 days in PBF2/3 treatment was associated with greater phosphate uptake in lettuce tissue. As regarding all results, PBF showed better lettuce growth compared to mineral phosphate fertilizer where PBF2/3 treatment resulted in increase of lettuce fresh weight by 23% and phosphate uptake by 50%.
Natural silk is formed by two proteins : the crystalline fibroin (inside the silk thread) and amorphous sericin (as a tube outside the thread). The degumming process is used to eliminate the external sericin prior to dyeing ; generally it makes use of soaps at about pH 10. Sericin is the protein constituent that "gums"together the fibroin filaments of cocoon silk. It constitutes about 25% of the weight of the cocoon, is soluble in hot water and "gels" on cooling. The removal of sericin from raw silk, known as degumming, is a simple but important process usually employing hot dilute soap or alkaline solution and occasionally dilute acids or enzymic methods. During degumming, alkali is taken up by the sericin and the free acid from the soap is formed ; this may be deposited on the fiber, reducing the rate of degumming and protecting it from hydrolysis. Alkali is often added to maintain or restore the pH of the baths, but it is rarely used alone, since it leaves the silk rather harsh in handle. If complete sericin removal is required as for printing, sodium carbonate may be added. If the pH of the bath exceeds 11, the fibroin is attacked. Recently, According to the development of electrolysis, we can be obtained the electrolytic reduction water(above pH 11.5) and electrolytic oxidation water (below pH 3). The aim of this work was to study a degumming process using electrolytic water and a possibility of sericin recovery. The new degumming process used electrolytic water operates at $95^\circ{C}$ for 2hr. without any reagents. The wastewater of this process are formed by a solution of sericin in water. This conditions suggest the study of a possible recovery of this protein (sericin) which has an amino acid composition suitable for many used in cosmetics, textile finishing agents, animal feeding, etc. The degumming process using electrolytic water is available to reduce treatment costs and pollute and at the same time to recover sericin.
Sulfur-utilizing autotrophic denitrification relies on an inorganic carbon source to reduce the nitrate by producing sulfuric acid as an end product and can be used for the treatment of wastewaters containing high levels of nitrates. In this study, sulfur-denitrifying bacteria were used in anoxic batch tests with sulfur as the electron donor and nitrate as the electron acceptor. Various medium components were tested under different conditions. Sulfur denitrification can drop the medium pH by producing acid, thus stopping the process half way. To control this mechanism, a 2:1 ratio of sulfur to oyster shell powder was used. Oyster shell powder addition to a sulfur-denitrifying reactor completely removed the nitrate. Using 50, 100, and 200 g of sulfur particles, reaction rate constants of 5.33, 6.29, and $7.96mg^{1/2}/l^{1/2}{\cdot}h$ were obtained, respectively; and using 200 g of sulfur particles showed the highest nitrate removal rates. For different sulfur particle sizes ranging from small (0.85-2.0 mm), medium (2.0-4.0 mm), and large (4.0-4.75 mm), reaction rate constants of 31.56, 10.88, and $6.23mg^{1/2}/l^{1/2}{\cdot}h$ were calculated. The fastest nitrate removal rate was observed for the smallest particle size. Addition of chemical oxygen demand (COD), methanol as the external carbon source, with the autotrophic denitrification in sufficiently alkaline conditions, created a balance between heterotrophic denitrification (which raises the pH) and sulfur-utilizing autotrophic denitrification, which lowers the pH.
Rateb, Sherif A.;Khalifa, Marwa A.;El-Hamid, Ibrahim S. Abd;Shedeed, Hesham A.
Asian-Australasian Journal of Animal Sciences
/
v.33
no.7
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pp.1068-1076
/
2020
Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa. Methods: In the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T0). The diluted specimens were stored at 4℃ for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T24, T48). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system. Results: The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa. Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.
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