• 미생물학회지
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• 제42권2호
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• pp.131-134
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• 2006

Clavicorona pyxidata DGUM 29005의 균사 생육을 위한 최적 배양조건 및 효소 활성을 조사하였다. 균사 생육을 위한 최적 온도 및 pH는 각각 $24^{\circ}C$ 및 5.0이었다. 사용된 복합배지 중 malt extract medium (MEM)에서 가장 좋은 균사 생육을 나타내었다. 최소배지로 Czapek-Dox 한천배지를 사용하고 탄소원으로 trehalose, mannitol, sucrose 및 maltose 첨가시 균사생육이 우수하였다. 전반적으로 무기질소원이 유기질소원 보다 균사 생육을 더 촉진하였으며, 무기질소원으로 calcium nitrate를 사용하였을 때 균사 생육이 가장 우수하였다. 인산원으로 $Na_2HPO_4$를 사용했을 때 균사생육이 촉진되었으며, 가장 우수한 비타민원은 p-aminobenzoic acid이었다. MEM 액체배지를 사용하여 $24^{\circ}C$에서 20일간 C. pyxidata DGUM 29005를 배양하여 균사외 분비효소 및 균사내 효소의 활성도를 측정한 결과, 균사외 분비 효소 중 laccase의 활성도가 다른 효소에 비해 높았으며, ${\alpha}$-amylase, chitinase, lipase 및 pretense의 활성도는 낮거나 없었다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.8-8
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• 2001

Mouse follicles require the addition of gonadotropins (Gns) to complete maturation and ovulation of oocyte and antrum formation of follicles in vitro. However, we tried examination of in vitro growth of mouse pre-antral follicles in medium without Gns and physiological factors. And also, pre-antral follicles were isolated from ovaries by mechanical method. Our present studies were conducted to evaluate on the growth of follicles and intra-follicular oocytes and antrum formation in vitro of mouse pre-antral follicles in two different media. Pre-antral follicles (91-120${\mu}{\textrm}{m}$) were isolated mechanically by fine 30G needles not using enzymes from ovary of 3-6 weeks old female ICR mice. Isolated pre-antral follicles were cultured in 20 ${mu}ell$ droplets of TCM (n=17; follicles: 107.8 $\pm$ 1.58 ${\mu}{\textrm}{m}$; oocytes: 59.9$\pm$1.2 ${\mu}{\textrm}{m}$) or MEM (n=12; follicles: 109.3$\pm$2.53 ${\mu}{\textrm}{m}$; oocytes: 55.4 $\pm$1.6${\mu}{\textrm}{m}$) under mineral oil on the 60mm culture dish. All experimental media was supplemented with 10% FBS but without Gns and/or physiological factors. Pre-antral follicles were individually cultured in drops for 8 days. Antrum formation and growth of pre-antral follicles and intra-follicular oocytes were evaluated using a precalibrated ocular micrometer at $\times$200 magnifications during in vitro culture. Results between different groups were analyzed using combination of Student's t-test and Chi-square, and considered statistically significant when P<0.05. Antrum formation of pre-antral follicles had started in two culture media on day-2. On day-8, antrum formation had occurred in 58.3%(7/12) of pre-antral follicles cultured in MEM, but only in 23.5% (4/17) of those cultured in TCM (P=0.0364). Growth of pre-antral follicles and intra-follicular oocytes were observed on day-4 and -8. On day-4, follicular diameters was similar (P=0.1338) in TCM (119.4$\pm$2.58 ${\mu}{\textrm}{m}$) and MEM (125.4$\pm$4.52 ${\mu}{\textrm}{m}$). However, on day-8, diameters of pre-antral follicles cultured in MEM (168.9$\pm$17.29 ${\mu}{\textrm}{m}$) was significantly (P=0.0248) bigger than that in TCM (126.7$\pm$4.28 ${\mu}{\textrm}{m}$). On day-4 and -8, diameters of intra-follicular oocytes were similar TCM (67.1$\pm$1.3 and 72.4$\pm$0.9${\mu}{\textrm}{m}$) and MEM (65.2$\pm$1.7 and 73.3$\pm$1.5 ${\mu}{\textrm}{m}$), respectively. We can conform that medium not supplemented with Gns and/or physiological factors can be used for in vitro antrum formation and growth of mouse pre-antral follicles and intra-follicular oocytes. In conclusion, MEM supplemented with FBS can be used for growth in vitro of mouse pre-antral follicles isolated mechanically.

• Asian-Australasian Journal of Animal Sciences
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• 제22권5호
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• pp.667-673
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• 2009

Ninety six intact male sheep (12 months old with mean live weight of about 35 kg) were used to assess the effects of restricted feeding on intake, digestion, nitrogen balance and metabolizable energy (ME). The animals were selected from two known Iranian small and large body size breeds: 48 Sangsari (S) and 48 Afshari (A), and were divided into two equal groups: restricted (R) and a control (C). Each group had 48 sheep (24 each breed). The experiment had a duration of 15 and 75 days adaptation and treatment periods, respectively. The animals were individually placed in metabolism cages and fed a diet based on pelleted concentrate mixture consisting of alfalfa, barley grain, cottonseed meal and barley straw. The animals in group C were fed ad libitum, while animals in group R were fed at maintenance level and maintained a relatively constant live weight. During the experiment, the average daily weight gain (ADG) of S and A animals in R group was 0.34 and -0.25 g/d (0.02 and -0.02 $g/kg^{0.75}/d$), respectively. While that of S and A animals in C group was 174.4 and 194.4 g/d (10.16 and 11.48 $g/kg^{0.75}/d$), respectively. Nitrogen (N) was determined by both measured and regression methods. Animals of R group stayed at about zero N balance (0.01 and -0.00 g $N/kg^{0.75}/d$ for S and A animals, respectively). The N retention of animals of both S and A breeds in C group were similar (0.45 and 0.46 g $N/kg^{0.75}/d$, respectively). Digestible organic matter intake (DOMI) and ME requirement for maintenance (MEm) were measured by both constant weight technique and regression method by regressing N balance on DOMI and ME intake on ADG. The measured DOMI during constant weight was 24.61 and 24.27 g $DOMI/kg^{0.75}/d$ and the calculated DOMI from regression equation was 24.24 and 24.22 g $DOMI/kg^{0.75}/d$, for S and A animals, respectively. The measured MEm was 402 and 401 kJ $ME/kg^{0.75}/d$ and the calculated MEm from regression analysis was 398 and 400 kJ $ME/kg^{0.75}/d$ for S and A breeds, respectively. There were no significant differences between both measured and regression techniques. There was no significant difference between S and A breeds for DOMI, N retention, MEm, digestibility and metabolizability values. Digestibility values for OM, GE and CP and metabolizability were significantly (p<0.05) higher in restricted feeding sheep compared with that of sheep fed ad libitum.

• 미생물학회지
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• 제40권2호
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• pp.100-103
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• 2004

Inonotus mikadoi IMSNU 32058의 최적 균사생장을 위한 배양학적 특성 및 세포효소 활성의 최적 조건을 조사하였다. 균사 생장을 위한 최적온도는 $27^{\circ}C$였고, 실험한 pH 범위중pH 4.5에서 최적균사 생장을 나타내었다. 사용된 복합배지 중 malt extract media (MEM)와Phellinus igniarius media (PIM)에서 가장 좋은 균사 생장을 나타내었다. 최소배지로써 Czapek-Dox 한천배지를 사용하여 탄소원으로 단당류, 이당류 및 다항류를 침가하였을 때 xylose, raffinose 및 carboxymethyl cellulose (CMC)에서 가장 좋은 생장을 나타내었다. 일반적으로 유기질소원이 무기질소원보다 생장을 더 촉진하였다. 유기질소원으로는 yeast extract, soytone, proteose peptone 및 bacto peptone을 사용하였을 때 균사생육이 우수하였다. 무기질소원으로는 ammonium sulfate를 사용했을 때 균사생육이 촉진되었으며, 가장 우수한 비타민원은 p-aminobenzoic acid (PABA)이었다. MEM 액체(pH 4.5) 배지를 이용하여 $27^{\circ}C$에서 5일간 배양하였을 때, I. mikadoi의 균사외 분비효소 및 균사내 효소의 활성도를 측정하였다. 균사내 분비 효소중 laccase 의 활성도가 다른 효소에 비해 상대적으로 높았다. 균사외 분비 효소도 균사내 분비효소와 마찬가지로 laccase의 활성도가 다른 효소에 비해 상대적으로 높았으며 protease, chitinase 그리고 lipase의 활성도는 낮거나 없었다.

• 한국수정란이식학회지
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• 제10권2호
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• pp.131-138
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• 1995

Essential and non-essential amino acids supplemented to culture medium stimulate mammalian embryo development in vitro. Amino acids such as glycine, taurine and alanine are concentrated in the lumen of oviduct and uterus and it can he thought that these amino acids may have physiological role on fertilization and embryo development. Our aim of this experiment was to investigate the effects of essential and non-essential amino acids, taurine or glycine supplemented to fertilization medium on the cleavage and subsequent in vitro development of bovine oocytes matured and fertilized in vitro. Immature oocytes were obtained from slaughtered Holstein cows and heifers and matured in TCM199 containing 10% fetal calf serum, 2.5 $\mu$g /mL of FSH and LH and 1 $\mu$g / mL of estradiol with granulosa cells in vitro. After maturation, oocytes were coincubated with sperm in fertilization medium supplemented with Minimum Essential Medium (MEM) essential and non-essential amino acids, taurine (3.75 mM) or glycine (10 mM) for 30 hours in vitro. Inseminated oocytes were cultured in synthetic oviduct fluid medium (SOEM) containing MEM essential, non-essential amino acids and 1 mM glutarnine up to 8 days after fertilization.Supplementation of fertilization medium with MEM essential and non-essential amino acids lowered significantly (p<0.05 and p<0.001) the cleavage rate after 30 hours of IVF (53.3%) and at Day 3 (62.7%: Day 0: the day of I VF) compared to control (64.3% and 77.3%, respectively). Subsequent developmental rates to morulae (Mo) and expanding blastocysts (ExBL) also significantly decreased (p<0.001 and p<0.05 for Mo and ExBL) when oocytes were coincubated with sperm in the medium containing MEM amino acids. Taurine added to fertilization medium have not increased the cleavage rate over the control, whereas glycine showed significantly lower (p<0.01) cleavage rate at Day 3 than that of taurine, but there was no significant difference in the developmental rates to Mo and ExBL of bovine embryos irrespective of the supplementation of taurine or glycine to fertilization medium. In conclusion, supplementation of fertilization medium with essential and non-essential amino acids, taurine or glycine has no beneficial effect on in vitro cleavage and development of bovine oocytes matured and fertilization in vitro.

• Journal of Periodontal and Implant Science
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• 제30권1호
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• pp.39-52
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• 2000

Polypeptide growth factor belong to a class of potent biologic mediator which regulate cell differentiation, proliferation, migration and metabolism. 1,25-dihydroxyvitamin $D_3$ decrease cell proliferation, and stimulate alkaline phosphatase activity which express in osteoblast during cell differentiation period. IGF-I is known to stimulate cell proliferation and differentiation too. 1,25-dihydroxyvitamin $D_3$ is known to increase IGF-I binding sites and IGF binding protein which inhibite the effect of IGF. The purpose of this study is to evaluate potential role of IGF-I as mediator that control the action of 1,25-dihydroxyvitamin $D_3$. MC3T3-E1 cell were seeded $5{\times}10^5/ml$ at 100mm culture plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 5% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ added. Total mRNA was extracted at 0, 6, 24, 48, 72 hour. PRPCR method was programed for the detection of IGF-I mRNA. In the both groups of 1,25-dihydroxy vitamin $D_3$ treated and control, alternative splicing form of IGF-I, IGF-IA and IGF-IB were expressed. In the 1,25-dihydroxyvitamin $D_3$ treated group, IGF-I mRNA expression was matained until 24 hour, there after expression was decresed. MC3T3-E1 cell were seeded $2.5{\times}10^4/ml$ at 24well plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 3% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ and 10 ng/ml IGF-I were added separately or together. Cell were cultured for 1 and 3 days, $2{\mu}Ci/ml\;[^3H]$ -thymidine was added for the last 24h of culture of each days. ${[^3H]}$-thymidine incorporation in to DNA was measured and expressed counter per minute(CPM). DNA synthetic activity was significantly decreased by 1,25-dihydroxyvitamin $D_3$ both at 1 day and 3 day, and in the combination group of 1,25-dihydroxyvitamin $D_3$ and IGF-I, DNA synthetic activity was also decreased both at 1 day and 3 days. IGF-I did not affect the DNA synthetic activity compared to control group both at 1 day and 3 day. From the above results, 1,25-dihydroxyvitamin $D_3$ was potent inhibitor of cell proliferaton in MC3T3-E1 cells. It assumed that the effect of 1,25-dihydroxyvitamin $D_3$ on osteoblast proliferation may be mediated in part by decreased level of IGF-I.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2008년도 추계학술대회 논문집
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• pp.31-32
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• 2008

• 한국광학회:학술대회논문집
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• 한국광학회 2004년도 하계학술발표회
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• pp.258-259
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• 2004

• 한국정밀공학회지
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• 제25권12호
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• pp.13-19
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• 2008

• 한국정밀공학회지
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• 제18권12호
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• pp.22-29
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• 2001