• 미생물학회지
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• 제43권4호
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• pp.292-297
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• 2007

신령버섯(Agaricus blazei Murill)의 액체 배양으로 다당체의 균사체외 분비를 유도하였으며, 버섯 균사체의 새포내 다당체와 세포외 분비 다당체의 면역증진활성을 in vitro 시험으로 비교하였다. 부분 정제된 세포내 다당체와 세포외 다당체의 총당 함량은 각각 85.6%와 95.3%였으며, ${\beta}$-glucan 함량은 각각 67.9%와 88.1%로 측정되었다. 면역활성 실험에는 시료의 닥 함량을 동일하게 맞추어 사용하였다. In vitro에서 균사체내외 다당체는 대식세포 주인 RAW 264.7을 활성화시켜 nitric oxide (NO) 생성을 농도 의존적으로 증가시켰으며, 각각 최대 53.9%, 53.1%의 비슷한 증가활성을 나타내었다. 또 균사채내외 다당체는 모두 RAW 264.7을 활성화시켜 염중성 cytokine류인 interleukin (IL)-$1{\beta}$, IL-6, tumor necrosis factor (TNF)-${\alpha}$의 생성을 증가시켰으며, 이때 3종의 cytokine 모두에서, 세포내 다당체에 비해 세포외 다당체를 처리했을때 저농도에서 높은 증가률을 나타내었다. 두 다당체는 in vitro 상에서 비장세포를 증식시키는 효과를 나타내었으며, 세포내 다당체가 농도 의존적으로 증식효과를 보인데 반해 세포외 다당체는 저농도에서 증식이 높았고, $250\;{\mu}g/ml$ 농도 이상에서는 더 높아지지 않았다. 두 다당체 모두 암세포인 B16F0 melanoma에 대한 직접적인 세포독성 효과는 나타내지 않았다. 신령버섯 균사체 배양으로 생성된 세포내외 다당체는 in vitro에서 모두 면역활성을 증가시키는 것으로 나타났으며 전반적으로 그 활성은 세포내 다당체보다 세포외 다당체가 우수한 것으로 판단되었다.

• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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• pp.43-50
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• 1998

This study was performed to determine the effects of nitric oxide on human sperm cell function. Semen samples were obtained from normal healthy volunteers. Motile spermatozoas collected by swim-up method were incubated up to 24 hours in Ham's F-10 medium supplemented with a various concentration of sodium nitroprusside (nitric oxide releasing agent). Sperm motility, hyperactivation, acrosome reaction rate, and acrosin activity were determined. The results are as follows; 1. 1mM of SNP resulted in a significant decrease in sperm motility ($44.8%{\pm}8.9%:78.1%{\pm}6.3%$, and hyperactivation $(10.4%{\pm}6.4%:47.7%:{\pm}9.5%)$ after incubation for 3 hours compared with the control group (Ham's F-10 alone), but had no effect on acrosome reaction. 2. At $100{\mu}M$ SNP, sperm motility was reduced after incubation for 6 hours $(54.8%{\pm}3.2%)$ compared with that of the control group $(82.7%{\pm}8.9%)$, but hyperactivation and acrosome reaction were not affected. 3. However, a lower concentration (less than $10{\mu}M$) of SNP had no effect on sperm motility and hyperactivation for 8 hours of incubation but significantly decreased them when incubation periods were increased up to 24 hours compared with the control group. On the other hand, $1{\mu}M$ and $10{\mu}M$ SNP significantly increased the acrosome reaction rate in both acrosomal status ($17.3%{\pm}5.2%$, $23.5%{\pm}4.7%$, respectively) and acrosin activity ($34.3{\mu}IU{\pm}10.5{\mu}IU,\;45.6{\mu}IU{\pm}5.6{\mu}IU$, respectively) as compared with the control group $(7.0%{\pm}4.0%,\;9.5{\mu}IU{\pm}3.4{\mu}IU)$. These results indicate that SNP, NO releasing agent, has a dose-dependent effects on the sperm cell function. Therefore it may positively affect the fertilization by promoting acrosomal reaction at a lower concentration (less than $10{\mu}M$).

• The Korean Journal of Physiology and Pharmacology
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• 제1권2호
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• pp.135-142
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• 1997

As it has been reported that the depolarization-induced norepinephrine (NE) release is modulated by activation of presynaptic $A_1$-adenosine heteroreceptor and various lines of evidence indicate that $A_2$-adenosine receptor also presents in hippocampus, and that the adenosine effect is magnesium dependent, the present study was undertaken to delineate the role of adenosine receptors in the modulation of hippocampal NE release. Slices from the rat hippocampus were equilibrated with $[^3H]-NE$ and the release of the labelled product, $[^3H]-NE$, was evoked by electrical stimulation (3 Hz, 5 V $cm^{-1}$, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium outflow was investigated. $N^6-cyclo-pentyladenosine$ (CPA), in concentrations ranging from 0.1 to 10 ${\mu}M$, decreased the $[^3H]-NE$ release in a dose-dependent manner without changing the basal rate of release, and these effects were significantly inhibited by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 2 ${\mu}M$) treatment. When the magnesium concentration was reduced to 0.4 mM or completely removed, the evoked NE release increased along with decreased basal rate of release. In contrast, increasing the magnesium concentrations to 2.4 and 4 mM, decreased the evoked NE release. The CPA effects on evoked NE release were reducedby magnesium removal, but potentiated by 2.4 mM magnesium in the medium. 5-(N-cyclopropyl)-carboxamodiadenosine (CPCA, 1 & 10 ${\mu}M$), an $A_2$-agonist, decreased the evoked tritium outflow, and this effect was also abolished by DPCPX pretreatment. CGS, a powerful $A_2$-agonist, did not affect the evoked NE release. However, the effects of CPCA and CGS on evoked NE release were significantly increased by pretreatment of DPCPX in the magnesium-free medium. These results indicate that inhibitory effect of $A_1$-adenosine receptor on NE release is magnesium-dependent, and $A_2$-receptor may be present in the rat hippocampus.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제22권2호
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• pp.155-163
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• 2000

Osteoporosis is the consequence of an imbalance between osteoclastic and osteoblastic activity, coupled with an increased rate of bone turnover observed with menopause. Estrogen is generally considered to maintain bone mass through suppression of bone resorption. The purpose of this study was to evaluate the rat femoral trabecular change not only in the deficiency of estrogen but also in the administration of estrogen following ovariectomy(OVX). 30 female Sprague-Dawley rats were subjected to bilateral OVX or sham surgery(control). Groups of OVX were divided into 4 groups. The first group was injected daily with vehicle alone for 20 days after 20 weeks following OVX. The additional groups of OVX was injected daily with low, medium, or high doses of $17{\beta}-estradiol$(10, 25 or $50{\mu}g/kg$ BW, respectively). All rats were sacrified 23 weeks after OVX, and their femur were processed for H&E, MT stain and histomorphometry. The results were as follows; 1. In the histomorphometric analysis, the trabecular bone volume/tissue volume, trabecular thickness and trabecular seperation were respectively $31.2{\pm}8.3%$, $54.3{\pm}4.8{\mu}m$ and $280.7{\pm}16.4{\mu}m$ in vehicle treated OVX group and $48.6{\pm}7.3%$, $90.4{\pm}4.5{\mu}m$ and $126.3{\pm}5{\mu}m$ in sham operation group, and they showed statistical significance compare to control group. 2. The trabecular bone volume/tissue volume, trabecular thickness and trabecular separation were respectively $44.4{\pm}4.3%$, $109.5{\pm}12.3{\mu}m$ and $94.9{\pm}8.5{\mu}m$ in low doses of $17{\beta}-estradiol$ injected group and they showed statistical significance compare to OVX group. 3. The trabecular bone volume/tissue volume, trabecular thickness and trabecular separation were respectively $44.4{\pm}4.3%$, $109.5{\pm}12.3{\mu}m$ and $94.9{\pm}8.5{\mu}m$ in medium doses of $17{\beta}-estradiol$ injected group and they showed statistical significance compare to OVX group, but they didn't show statistical significance compare to low doses of $17{\beta}-estradiol$ injected group. 4. The trabecular bone volume/tissue volume, trabecular thickness and trabecular separation were respectively $46.4{\pm}4.5%$, $154.4{\pm}13.2{\mu}m$ and $113.7{\pm}12.8{\mu}m$ in high doses of $17{\beta}-estradiol$ injected group and they also showed statistical significance compare to OVX group, but they didn't show statistical significance compare to other experimental groups. From the above results, metaphyseal bone formation was markedly reduced in OVX rate but treatment of OVX rats with $17{\beta}-estradiol$ resulted in normalization of femur trabecular bone volume. But they didn't show statistical significance the effect of bone formation according to the dose dependency.

• 한국수정란이식학회지
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• 제28권3호
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• pp.251-256
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• 2013

The present study was performed to investigate the effect of Hh-Ag1.5, a small-molecule chemical agonist of SMOothened receptor, on the in vitro maturation and development of in vitro fertilized (IVF) embryos in pigs. Oocytes or fertilized embryos were cultured in a maturation or embryo culture medium supplemented with 0 (control), 25, 50 or 100 nM of Hh-Ag1.5, respectively. Although the maturation rate were not different among treatment groups, the blastocyst formation rate in the group treated with 25 nM Hh-Ag1.5 was significantly increased compared to other groups (P<0.05). While the highest dose of Hh-Ag1.5 (100 nM) did negatively affect to the embryo development and cell number in blastocysts compared to other groups (P<0.05), the apoptotic cell index in blastocysts was significantly lower in 25 and 50 nM groups than in control and 100 nM groups (P<0.05). The mRNA expression of the proapoptotic gene Bax and the ratio of Bax/Bcl-XL decreased in among treatment groups compared to control (P<0.05). The embryo quality related genes, Tert and Zfp42, were significantly decreased in 50 and 100 nM groups compared with control and 25 nM groups (P<0.05). In conclusion, the addition of 25 nM Hh-Ag1.5 to in vitro maturation and culture medium can enhance the developmental potential as well as quality of IVF embryos in pig.

• 한국가축번식학회지
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• 제21권2호
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• pp.147-156
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• 1997

Ultrasound-guided follicular aspiration was performed in Holstein heifers once weekly with or without pretreatment of single or multiple decreasing doses using a total of 400 mg FSH. Oocytes were aspirated with a 6.5 MHz convex-array ultrasound trasducer designed for intravaginal use. All the visible follicles larger than 4 mm in diameter were punctured with a 17 gauge, 55 cm needle at each aspiration session and the follicular fluids containing oocytes were obtained by vacuum suction. The results obtained were as follows: As a preliminary experiment, the recovery rates of folicular oocytes by ultrasound-guided aspiration from the isolated ovaries of Korean native cows were compared between suction methods using manual syringe or vacuum pump. The recovery rate of oocytes using vacuum pump (80.7%) was significantly (P<0.05) higher than that using manual syringe (47.1%). The follicles were counted by their size in diameter with ultrasound image, and recovery rates and grades of follicular oocytes collected by ultrasound-guided aspiration were investigated in Holstein heifers pretreated with or without FSH. A group of heifiers were injected with multiple decreasing doses (twice a day for 3 days) of a total of 400 mg FSH. The other 2 groups were injected with a single dose of 400 mg FSH mixed with 25% PVP. Ultrasound observation of follicle population and/or ultrasound-guided transvaginal oocyte aspiration were performed 12 hrs following the last FSH injection in the multiple dose group, and 48 or 60 hrs after FSH injection in the single dose groups. Most of the visible follicles had small size of less than 3 mm in diameter in unstimulated heifers (71.0%), but medium size in all the heifers treated with FSH. (70.5 to 92.8%). The number of OPU follicles per session (4.6$\pm$1.9) were much less, compared to the vilsible follicle counts (9.7$\pm$2.2), in the nustimulated heifers due to the small dominant follicles. Among 4 goups of heifers the most visible as well as OPU follicles were observed in the heifers at 60 hrs following treatment of a single dose of 400 mg FSH (21.2$\pm$2.3 and 21.0$\pm$2.0), and the differences in both the follicle counts between the groups was found significant (P<0.05) The rates of oocyte recovery from the follicles by ultrasound-guilded aspiration were varied 46.3 to 75.0% in the heifers unstimulated and treated with a single dose of 400 mg FSH, but the group difference was not significant. The number of recovered oocytes per session a, pp.ared to be highest at aspiration at 60 hrs following single FSH (10.6$\pm$2.2) than at aspiration at 48 hrs after single FSH (7.8$\pm$2.7) or in the unstimulated heifers (3.4$\pm$3.0). The proportion of grade I and II oocytes to all oocytes collected was varied 31.8 to 64.0% between the groups. However, there was found no significant difference in both the number of oocytes recovered per session and the percentage and the percentage of grade I and II oocytes. From the above results it was concluded that the more oocytes of superior quality might be recovered economically by ultrasound-guided aspiration at 60 hrs following the pretreatment of a single dose of 400 mg FSH and by suction using a vacuum pump system of about negative pressure of 75 to 85 mmHg.

• Mycobiology
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• 제30권3호
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• pp.160-165
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• 2002

Aspergillus niger has been used as a host to express many heterologous proteins. It has been known that the presence of an- abundant protease is a limiting factor to express a heterologous protein. The protease deficient mutant of A. niger was obtained using UV-irradiation. A total of $1{\times}10^5$ spores were irradiated with $10{\sim}20%$ survival dose of UV, 600 $J/m^2$ at 280 nm, and the resulting spores were screened on the casein-gelatin plates. Ten putative protease deficient mutants showing the reduced halo area around colonies were further analyzed to differentiate the protease deficient mutant from other mutant types. Among ten putative mutants, seven mutants showed significant growth defect on nutrient rich medium and two mutants appeared to be the secretory mutants, which resulted in the impaired secretion of extracellular proteins including proteases. A mutant $pro^--20$ showed reduced halo zone without any notable changes in growth rate. In addition, the starchdegrading and glucose oxidase activities in the culture filtrate of $pro^--20$ mutant showed the similar range as that of the parental strain, which suggested that the $pro^--20$ mutant ought to be the protease deficient mutant rather than a secretory mutant. The reduced proteolytic activity of the $pro^--20$ was demonstrated using SDS-fibrin zymography gel. The reduced extracellular proteolysis was quantified by casein degradation assay and, comparing with the parental strain, less than 30% residual extracellular protease activity was detected in the culture filtrate of the $pro^--20$ mutant. The bio-activity of an exogenously supplemented hGM-CSF(human Granulocyte-Macrophage Colony Stimulating Factor) in the culture filtrate of $pro^--20$ mutant was detected until eight times more diluted preparations than that of the parental strain.

• 한국약용작물학회지
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• 제14권6호
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• pp.371-375
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• 2006

To investigate effects on the growth and ginsenosides accumulation in ginseng hairy root, potassium phosphate was supplemented with 1.25, 2.5, and 5.0 mM concentration in 1/2 MS medium, respectively. Potassium phosphate supplement was increased the biomass and ginsenosides accumulation when it was dose at the concentration of 1.25 mM. And the growth rate of hairy root in the light condition was higher than in dark condition. The highest contents and productivity of ginsenosides were observed at the supplement of 1.25 mM potassium phosphate at the 7th day after the culture onset. Ginseng hairy root cultured in 20 L bioreactor supplemented with 1.25 mM KH$_2$PO$_4$ was increased the growth with 1,609 g (F${\cdot}$W) and ginsenosides content with 11.09 mg than those in control.

• Journal of Pharmaceutical Investigation
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• 제27권4호
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• pp.313-321
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• 1997

Ion exchange resin complexes of famotidine have been prepared by the reaction of famotidine solution with activated ion exchange resins. Complex formation efficiency between famotidine and ion exchange resin was about $80{\sim}90%$ in average, calculated by HPLC determination. Drug release characteristics from the resin complexes were evaluated by the modified percolation method. Famotidine release was dependent on the type of ion exchange resins. In the case of weakly acidic resin complexes, the cumulative released amount of famotidine was more than 90% for 1hr in pH 1.2 buffer solution. However, in the case of strongly acidic resin complexes, it was less than 5% for 3hr in the same medium. Strongly acidic resins revealed some advantages over weakly, acidic resins for overcoming instability of famotidine in gastric juice. In addition, strongly acidic resin complexes showed controlled release of famotidine in pH 6.8 buffer solution, showing the result of about 60 to 70% of drug release for 5hr. After oral administrations of famotidine-resin complexes to rats as dose of 40 mg equivalent/kg, the pharmacokinetic parameters of famotidine were obtained by model independent analysis and compared with those of famotidine solution or suspension. $C_{max}$ of famotidine-resin complex was lower than that of famotidine solution or suspension. MRT, MAT, and MDT of the complexes were greater than those of famotidine solution or suspension. From these results, it was expected that famotidine was released slowly from the complexes and absorbed continuously into systemic circulation. It was recognized that drug release from the complexes was the rate-limiting step in drug absorption, since there were close correlations between in vitro drug release and in vivo pharmacokinetic parameters.

• 식물조직배양학회지
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• 제22권6호
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• pp.339-343
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• 1995

철이온이 Arabidopsis thaliana의 mesophyll protoplasts 배양에 미치는 영향을 조사하기 위하여 mesophyll protoplasts를 분리한 후 변형시킨 IMH 배지에 서로 다른 농도의 Fe-EDTA를 조성하여 배양하는 동안 나타나는 현상들을 관찰하였다. 세포분열률, 미세캘러스의 생장 및 첫번째 세포분열의 시기 등이 Fe-EDTA 농도에 따라 현저히 달라짐을 확인하였다. 대조군에서는 전혀 세포분열이 관찰되지 않았으며, 0.02 mM 미만의 저농도에서는 세포분열률이 낮았다. 0.5-1 mM에서 가장 빠른 세포분열과 높은 분열률을 보였으며 미세캘러스의 형성과 생장도 가장 빨랐다. 낮은 농도(0-0.25 mM)의 Fe-EDTA가 첨가된 배지에서는 농도의 증가와 비례하여 분열률이 높아졌고, 첫 세포분열의 시기와 미세캘러스의 생장 또한 농도의 증가에 비례하여 촉진되는 것으로 나타났다. 그러나 이러한 현상은 1 mM에서최고치를 보인 후 그 이상의 농도에서는 농도의 배가에 따라 효율이 현저히 감소되는 것으로 관찰되었다. 본 연구에서 엽육세포 원형질체의 세포분열 및 미세캘러스의 생장이 최대로 나타났던 0.5-1 mM 철이온 농도는 현재 일반적으로 사용되고 있는 배지들에 비해 5-10배의 높은 농도이다.