• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2005년도 2005 Annual Meeting & International Symposium
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• pp.319-322
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• 2005

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2008년도 제49회 학술심포지움 및 국제학술대회
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• pp.78.2-78.2
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• 2008

• 한국동물학회:학술대회논문집
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• 한국동물학회 2001년도 공동 춘계학술대회
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• pp.78-78
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• 2001

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• Journal of Plant Biology
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• 제30권3호
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• pp.201-203
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• 1987

Total RNA was isolated from two months old wheat, rice, tobacco and sweet potato. The procedure used was simple and provided pure RNA preparation. Lysis of plant tissue in a buffer with guanidine thiocyanate and CsCl density gradient centrifugation separated RNA from the rest of the cellular components. Subsequent cholroform/1-butanol extraction and ethanol precipitation were necessary to ensure contaminant-free RNA preparation. Storage of the lysed plant tissue in the buffer with guanidine thiocyanate preserved the sample for two months without noticeable RNA degradation.

• Food Science and Biotechnology
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• 제17권5호
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• pp.925-930
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• 2008

Norovirus has become the most common cause of human gastroenteritis in developed countries. Detection procedures of foodborne viruses from foods require several steps. The concentration step using polyethylene glycol (PEG) is time-consuming and the detection efficiency of reverse transcription-polymerase chain reaction (RT-PCR) is affected by inhibitors from food components. In this study, a rapid detection method based on buoyant density centrifugation was developed to replace the time-consuming chloroform-polyethylene glycol-Tris Tween method. Feline calicivirus that belongs to the family Caliciviridae was used as a surrogate model for norovirus. After artificial inoculation of feline calcivirus (FCV) to oyster and lettuce, 830 ${\mu}L$ of homogenized sample suspension was layered on the top of 670 ${\mu}L$ 20% percoll and centrifuged. Then RNA extraction step was proceeded with the supernatant. By varying several physical conditions, the detection limits were lowered to $2.4{\times}10^2$ PFU per 1 g in oyster and $2.4{\times}10^0$ PFU per 1 g in lettuce. The protocol obtained in this study could be used to develop new detection method for norovirus in foods.

• 대한미생물학회지
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• 제34권5호
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• pp.423-433
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• 1999

A novel bacteriophage, designated as VPP97, that infects the strains of Vibiro parahaemolyticus (hallophilic, Gram-negative bacterium) isolated most commonly from marine environments, has been discovered, and several of its properties have been determined. The plaques were clear and sized $0.6{\sim}1.0\;mm$ in diameter. The virion forms a single band on 70% sucrose gradient and ${\rho}1.50$ CsCl gradient by sucrose gradient centrifugation and CsCl gradient centrifugation respectively. It has a hexagonal head and a relatively long tail, as shown by electron microscopy. Vibrio alginolyticus, Vibrio fluvialis and Vibrio furnissii were also sensitive to this phage. It was almost totally inactivated at $70^{\circ}C$ and at pH below 5 or over 10. The nucleic acid of VPP97 is composed of DNA. The VPP97 had 9 specific structural proteins sized between 21.5 kDa and 97.4 kDa on SDS-PAGE. When V. parahaemolyticus cultures were treated with either phage VPP97 or one of the several antibiotics for 2 hours, the viable number of V. parahaemolyticus treated with the phage VPP97 is lower than that treated with chloramphenicol, erythromycin or penicillin, but not lower than that treated with tetracycline. Mice that have responded to the phage treatment revealed the lower numbers of V. parahaemolyticus in small intestine and less damage on small intestine compared to the untreated mice. Therefore, we suggest that the phage treatment appears effective to the infection by V. parahaemolyticus.

• Archives of Plastic Surgery
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• 제42권2호
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• pp.143-149
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• 2015

Background Adipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at $-20^{\circ}C$ by using thawing methods used in clinics. Methods The survival rates of adipocytes in the following thawing groups were measured: natural thawing at $25^{\circ}C$ for 15 minutes; natural thawing at $25^{\circ}C$ for 5 minutes, followed by rapid thawing at $37^{\circ}C$ in a water bath for 5 minutes; and rapid thawing at $37^{\circ}C$ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes. Results In the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05). The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant. Conclusions It was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at $37^{\circ}C$. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.

• 한국토양비료학회지
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• 제45권1호
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• pp.30-36
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• 2012

The active fraction of soil organic matter, which includes organic debris and light organic fraction, plays a major role in nutrient cycling. In addition, particulate organic matter is a valuable index of labile soil organic matter and can reflect differences in various soil behaviors. Since soil organic matter bound to soil mineral particles has its density lower than soil minerals, we partitioned soil organic matter into debris ($<1.5g\;cm^{-3}$), light fraction ($1.5-2.0g\;cm^{-3}$), and heavy fraction ($>2.0g\;cm^{-3}$), based on high density $ZnCl_{2-}$ sucrose solutions. Generally, partitioned organic bands were clearly separated, demonstrating that the $ZnCl_{2-}$ sucrose solutions are useful for such a density gradient centrifugation. The available gradient ranges from 1.2 to $2.0g\;cm^{-3}$. Although there was not a statistically meaningful difference in organic debris and organomineral fractions among the examined soils, there was a general trend that a higher content of organic debris resulted in a higher proportion of light organomineral fraction. In addition, high clay content was associated with increased fraction of light organomineals. Partitioning of soil organic carbon revealed that carbon content is reduced in the heavy fraction than in the light fraction, reflecting that the light fraction contains more fresh and abundant carbon than the passive resistant fraction. It was also found that carbon contents in the overall organic matter, debris, light fraction, and heavy fractions may differ considerably in response to different farming practices.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2006년도 제23차 정기총회 및 학술발표회
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• pp.69-72
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• 2006

To investigate the manufacturing methods of surimi-like materials (SLM) from breast muscle of spent hen, the muscles were diced, chopped and washed with distilled water or sodium chloride solution at 0.1, 0.5 and 1% level and then washed with distilled water to extract myofibrillar protein. When used only distilled water to extract myofibrillar protein, washing was repeated 3 times followed by homogenization and centrifugation of breast muscle after each washing (CM; conventional method). Whereas, to extract myofibrillar protein using sodium chloride solution had sufficient to do 2 times washing by distilled water after 1 time washing by sodium chloride followed by homogenization and centrifugation of breast muscle after each washing (NM; new method). The both batter and cooked SLM gel from NM had significantly (p<0.05) lower redness compared with CM. Again, SDS-PAGE with sarcoplasmic protein fractions showed that the bands of phosphorylase had increased staining intensity in NM compared with CM. These results indicated that the brightness was related to sarcoplasmic protein fractions. SDS-PAGE with myofibrillar protein showed that the bands of myosin had increased staining intensity in NM compared with CM. Data implied that myofibrillar protein extraction with sodium chloride solution had the better adaptability for the breast muscle of spent hen then the commonly used distilled water method.

• 한국동물생명공학회지
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• 제35권2호
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• pp.142-148
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• 2020

The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode's albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.