• Journal of Veterinary Clinics
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• v.22 no.4
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• pp.377-381
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• 2005

We tried to extract bone morphogenetic protein (BMP) from the freeze-dried bovine cortical bone (FBCB) for bone graft, which were defatted with chloroform-methanol for 20 days, freeze-dried at $-80^{\circ}C$ for 7 days and sterilized by ethylene oxide gas. Two kg of FBCB were pulverized in a wheel mill to $0.5-2.0mm^3$ cubic in size. The bone particles were demineralized in 0.6N HCI for 10 days at chloroform-methanol$4^{\circ}C$ and defatted with chloroform-methanol for 6 hours at room temperature, which was going to be defatting and demineralized cortical bone (DDM). For extracting BMP, DDM was agitated continuously through 72 hours with magnetic stirrer at $4^{\circ}C$ into 12 times of volume of 6 M guanidine hydrochloride (Gdn-HCl) solution containing proteinase inhibitors to protect BMP such as 2mM N-ethylaleimide, 1mM iodoacetic acid, 1mM phenylmethylsulfonyl fluoride and a sterilizer, 1mM sodium azide. The extraction procedure was repeated for three times. All extracted solution was centrifuged at 10,000 rpm for 30 min and then, the supernatant was dialyzed with 12 times of volume of deionized water at $4^{\circ}C$ for 24-72 hours, which cut off below 6,000-8,000 molecular weight. The dialyzed specimen contained crude-BMP was centrifuged, freeze-dried, and weighted. Through these processing, we could obtained $84.9\%$ as FBCB, $17.8\%$ as DDM and $0.71\%$ as crude-BMP from the wet cortical bone without cancellous bone, marrow and muscles. The crude-BMP were obtained $68.3\%$ from the first extraction, $29.6\%$ from secondary and $2.1\%$ from tertiary, respectively. It was suggested that high yield of crude-BMP migth be explained by three-time repetition of the extraction processing for crude-BMP with Gdn-Hcl sol.

• Journal of radiological science and technology
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• v.30 no.3
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• pp.259-263
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• 2007

For the preparation of $^{99m}Tc$-labeled RBC, $10{\sim}20\;{\mu}g/kg$ of Stannous(II) chloride and $10{\sim}40\;min$ of preparation was used. For finding out the effect of contrast agent, the blood samples were collected in three days, seven days, and 1 months after the diagnostic procedure. In the normal volunteer, the concentration of reducing agent and preparation time did not effect on the radiochemical yield. But in the patients, 10 mg of Stannous(II) chloride and 60 min incubation times was shown high radiochemical yield. Contrast agent has a significant effect on the radiochemical yield. Although the blood samples which were collected after seven days of diagnostic procedure did not effect on the radiochemical yield of $^{99m}Tc$-labeled RBC, but the radiochemical yield of $^{99m}Tc$-labeled RBC which was prepared with a sample of high concentration of contrast agent in blood led to low radiochemical yield. For these samples, the modified method showed high radiochemical yield than previous in vivo preparation method. The recommended method is followed. Blood collecting was performed at 30 minutes after injection of reducing agent, and it is centrifuged for removal of plasma. After addition of $^{99m}TcO^-_4$, sample reservoir was rotated. After addition of normal saline, and it is centrifuged for separation of saline. Then $^{99m}Tc$-labeled RBC was obtained after removal of saline.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.181-189
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• 2001

The objective of this study was to evaluate the development of porcine follicular oocytes fertilized by intracytoplasmic sperm injection (ICSI). Cumulus-oocyte-complexes (COCs) were collected by aspiration from follicles of 2~7 mm in diameter from a local slaughterhouse. Oocytes were matured in vitro for 40~44 h, and spermatozoa were prepared by swim-up in the presence or absence of 5 mM dithiothreitol (DTT) and then M II stages of the oocyte were either centrifuged or not centrifuged for the following injection of ooplasm. Injected oocytes were cultured in NCSU 23 medium for 6 to 8 days. The results obtained were as follows: 1. The rates of cleavage and development rates into blastocyst by ICSI were not significantly different between the with (53.0% and 19.7%) or without (48.3% and 23.8%) centrifugation, respectively (P＜0.05). 2. The cleavage and developmental rates to blastocyst after ICSI with or without 5 mM DTT treated-sperm were not significantly different (60.4% vs 16.4% and 45.5% vs 22.2%), respectively (P＜0.05). 3. The cleavage and the developmental rates to btastocyst were not significantly different between the zygotes obtained by IVF (51.8% vs. 22.4%) and ICSI (51.4% vs. 21.6%) (P＜0.05). 4. The number of blastomere in blastocyst stages after IVF or ICSI was not significantly different (46.7$\pm$2.9 and 41.9$\pm$4.6).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.16 no.2
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• pp.125-132
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• 1983

Dielectric properties of sardine-starch paste with moisture content of 4 to $13\%$ were investigated as functions of moisture and frequency. And the effects of the levels of fat and starch of the mixtures upon dielectric permittivity, critical moisture, were also mentioned. In addition, a theoretical prediction of frequency dependence of dielectric constant which was computed by the lumped circuit of two layer condenser model was evaluated. For the preparation of sardine-starch paste, comminuted sardine meat was washed thoroughly several times in chilled water by soaking and decanting, and finally centrifuged. This procedure was extended longer to provide a low fat sample. The centrifuged meat was mixed with adequate amounts of starch and salt, and ground for 25 minutes in a stone mortar, moulded in the form of disk with 7cm diameter and 1.2cm thickness and then freeze dried. Dried meat disks were cut off for the size of 5.5cm diameter and 1.0cm thickness and their moisture contents were controlled in humidified desiccators with saturated solutions. Dielectric constants of sardine-starch paste tended to decrease frequency was increased showing a critical charge at the moisture called critical moisture content. In case of the sample with $20\%$ starch and $2\%$ salt an average complex permittivity($\epsilon^{\ast}$) at 7 to $8\%$ morsture as the critical moisture content was presented; $\epsilon^{\ast}$=3.37+j 0.39 at 0.1 MHz, $\epsilon^{\ast}$=2.54+j 0.19 at 15 MHz, and $\epsilon^{\ast}$=2.15+j 0.08 at 1.8 GHz, respectively. The theoretically obtained complex permittivity values from the two layer condoner model were in close agreement with these actual measurements under the same conditions, that appeared as $\epsilon^{\ast}$=2.53+i 0.09 at 0.1 MHz and $\epsilon^{\ast}$=2.28+j 0.06 at 15 MHz, respectively. The fast level of the mixture also revealed an influence on dielectric property that defatted neat with $1.0\%$ fat showed a higher hc and $\epsilon^{\ast}$ value than the meat with $4.8\%$ fat. Complex permittivity being related to the moisture level remained nearly unchanged or slightly changed at the moisture range of 4 to $8\%$ but was dispersed widely at higher moisture contents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.3
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• pp.461-466
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• 2010

A cylindrical UV sterilization system was developed to decrease microorganisms in centrifuged Takju (CT). CT was run through 110 strips of honey comb type-teflon tubes and 9 UV lamps (1395 W) were equipped between teflon tubes. The optimum sterilization condition of CT was fixed for 1.5 min at 2 L/min in overall quality aspects; also, 5~6 log cycle decrease of viable cell numbers of total bacteria and yeast was observed at this operating condition. Quality changes of UV-sterilized CT were examined by UV irradiation of CT followed by storing at $30^{\circ}C$ for 8 days. To evaluate quality changes of UV-sterilized CT, pH, amino nitrogen content, acidity, reducing sugar content and viable cell numbers of total bacteria and yeast were measured. The growth of yeast and bacteria was retarded, showing around $10^8\;CFU/mL$ even after 4 days and $10^8\;CFU/mL$ after 6 days, respectively. Also, UV sterilized CT showed no changes in pH, titratable acidity, and amino nitrogen content during storage except reducing sugar content. UV sterilization did not cause significant difference in L, a, and b values between CT and UV-sterilized CT over the storage period.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.128-131
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• 1996

Lactobacillus acidophilus KFRI 233 (of human origin) exhibited a high tolerance to bile. The maximum cell yield was 6.6${\time}10^9 CFU$ per gram of whey in a 5.0% whey medium. Cell growth was improved with the addition of 0.5% thiotone and 0.25% calcium carbonate. Cell growth reached a maximum level of 5.4${\times}10^8$ CFU/ml at 20 h. Eighty-nine percent of the viable cells in the centrifuged concentrate survived freezing at $-70^{\circ}C$ and this frozen concentrate showed no reduction in the viable cell count after 30 days at $-70^{\circ}C$. Eight percent of the viable cells survived freeze-drying after the addition of 1 g/l sodium carbonate before harvesting by centrifuging and this freeze-dried concentrate showed only a slight reduction in the viable cell count after 30 days at $4^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.82-88
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• 1997

To produce biopolymer, Metarrhizium anisopliae (Metschn.) Sorok was cultured in a medium containing glucose 1.0%, sucrose 2.0% , soluble starch 1.0%, yeast extract 0.5%, polypeptone 0.05%, K$_{2}$HPO$_{4}$ 0.1% MgSO$_{4}$ $\CDOT $ 7H$_{2}$O 0.02%. The culture broth was centrifuged and the polymer was harvested by adding methanol to the culture supermatant. When three times of methanol was added, the polymer was coagulated and precipitated. Then it was further purified through successive SK-1B, SA-20P, HP-20 column chromatographies. This polymer was designated as Biopolymer YU-122.C:H ratio of this Biopolmer YU-122 was 1:2 and small amount of N is detected by CHN analyzer. Glucose and glactose are main components of this polymer. Average molecular weight of this biopolymer was 1.7%$\times $10$^{6}$ by Sepharose 4B gel permeation chromatography. Optimal condition for biopolymer production was investigated. When 5% of mannitol was used as a carbon source, and polypepton as a N source, highest productivity of biopolymer was achieved. C/N ratio as nutrient was also a major factor in polymer production and its optimal ratio was 3.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.522-524
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• 1999

A discontinuous Percoll gradient was used to separate islets from the collagenase-treated mouse pancreas easily and rapidly. Since the osmolality of Percoll is very low, adjustment of its osmolality to 340 mOs/kg $H_2$O was essential for securing the optimal separation. A discontinuous gradient layering with Percoll solution of 1.09 g, 1.07g, and1.05g/m, respectively, when centrifuged at 800$\times$g for 10 min, resulted in an optimal condition for separation and yielded a banding pattern with an even distribution of islet cells. No significant difference was observed in the morphological features between the Percoll-isolated and the manually-isolated islets. In conclusion, the discontinuous Percoll gradient can be effectively used to isolate the pancreatic islets from mice with four-fold higher efficiency compared to the handpicking method.

• Toxicological Research
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• v.11 no.1
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• pp.31-36
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• 1995

Cyclophosphamide (CP) must be enzymatically activated by cytochrome P450(CYP)-linked mixed-function oxidation pathway to be either mutagenic or teratogenic. Influences of alterations in hepatic mixed-function oxidase acitivity and glutathione (GSH) content on the embryotoxicity of CP were studied in rat whole embryo culture system. The embryotoxicity of CP was compared using rat S-9 fraction (S-9) pretreated with chemicals inducing different CYP isozymes, acetone (ACE), Aroclor 1254 (ARO), $\beta$-naphthoflavone (NAF) and phenobarbital (PHE). When 10.5 day embryos were cultured in the immediately centrifuged rat serum for 48 hrs using general gas char{ging schedule, CP$(40{\mu}g/ml)$ with S-9 induced by either NAF or PHE increased the incidence of realformations and significantly decreased embryonic growth compared with the non-induced S-9 group. ACE or ARO induced S-9 group showed no significant difference in embryonic growth. These data suggest that PB and/or NAF inducible CYP isoenzymes are mainly involved in the activation of CP. To examine the effect of GSH on the embryotoxicity of CP, 10.5 day embryos were exposed to CP and S-9 after preincubation with 10 mM of GSH for 3 hrs. In the GSH pretreated group the growth of embryos increased significantly compared with that of the untreated group, suggesting that GSH may protect embryos in culture from some toxic effects of CP.

• Journal of Korean Medicine for Obesity Research
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• v.11 no.1
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• pp.35-46
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• 2011

Objectives This study was designed to examine antimicrobial effects of Fermented Coptidis rhizoma and Lonicerae Flos against pathogens. Methods Lactobacilli MRS broth was added to 200mL glass bottle containing 20% herb powder(w/v) followed by 30 minute sonication and then shaking at 70 rpm in $70^{\circ}C$ water bath for 3 hours in order to extract fermented herb. Fermented herb extract was autoclaved at $121^{\circ}$ for 15minutes. $2{\times}10^7$ CFU/mL subcultured bacteria was inoculated and cultured for 24 hours and centrifuged at 3000 rpm for 5 minutes. After transferring to 15 mL conical tube, the viable cells were counted. Results and Conclusion Fermented Coptidis rhizoma and Lonicerae Flos both showed antimicrobial effect on pathogens especially when Fermented Coptidis rhizoma was experimented against Staphylococcus aureus.