• 아동학회지
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• 제33권1호
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• pp.41-62
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• 2012

This study explored the case of sandplay therapy for a 4th grade girl suffering from selective mutism. Her selective mutism apparently began following an extremely embarrassing experience in kindergarten. Her symptoms were a combination of symbiotic, reactive and passive- aggressive type behaviors. The goal of the therapy undertaken with this child was to enable her to express her repression and suppression, within a free and protective space during sandplay therapy. There were a total of 60 sessions of sandplay therapy. The client described the situations she had experienced in the first sandtray, by placing babies absent caring adults and food on the sandbox. She also placed baby fish away from their mother. In the mid-point of the sessions, she repeated her regressive behaviors by babbling like a baby and fought with snakes and monsters. In the final sessions, she showed integration and adaptation by engaging in snow play, expressing the union of opposites, placing blue and red mig and making a cross on the sand. This study showed the effectiveness of sandplay therapy since her selective mutism was lessened.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권6호
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• pp.419-425
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• 2001

A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.259-268
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• 1993

폐흡충의 분비배설물은 항원성이 높아 폐흡충증의 진단용 항원으로 가치가 있다고 보고되었다. 이 까닭은. 분비배설물에는 숙주내에서 여러 생물학적 반응을 일으키는 물질뿐 아니라 충체의 일부구성 성분 및 여러 효소 등이 포함되어 있기 때문이라 가정할 수 있다. 이 연구는 폐흡충 성충의 분비배설물에서 숙주의 산소라디칼을 분해시키는 효소인 catalase, superoxide dismutasr (SOD), peroxidase 등이 존재하는지를 확인하고 그 활성도를 측정하였으며 그 중 peroxidase를 정제하여 생화학적 특성의 일부 및 항원성을 관찰하였다. 폐흡충 성충 50마리를 $37^{\circ}C$ 부란기에 12시간 배양한 뒤 배양액을 원심분리하고 이를 분비배설 조효소로 사용하였다. 분비배설물에서 catalase, SOD와 peroxidase의 활성도를 측정할 수 있었고 그 비활성도(specific activity)는 각각 11.1, 3.4 및 48.5 이었다. 분비배설물의 peroxidase 비활성도는 충체추출액의 비활성도보다 1.5배 높았다 이 효소를 Sephacryl 5-300 Superfine gel filtration. DEAE-Tnsacryl M anion exchange chromatography로 정제하였다. 정제한 peroxidase의 분자량은 HPLC에서는 19 kDa이었고 SDS-전기영동에서는 16 kDa이었다. 정제한 효소를 전기영동한 후 diaminobenzidine으로 specific staining한 결과 이 효소는 충체추출액의 효소와 같은 영동이동거리를 나타내었다 즉 이 효소는 충체로부터 분비되는 것임을 알 수 있었다. 한편 폐흡충 감염 환자의 혈청과 반응시킨 immunoblot에서 분비배설물의 구성 단백질중 84, 64, 42, 32, 30, 28, 26, 24, 22, 11 및 8 kDa가 항원성을 보인 반면 정제한 peroxidase는 미약한 반응을 보였다 이상의 결과로 폐흡충 peroxidase는 분비배설되어 SOD, catalase와 함께 산소 독성을 제거하는데 작용하고 있으나 패흡충 감염 환자에서 특이 항체 반응을 일으키는데 에는 미약한 작용을 한다는 것을 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제27권6호
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• pp.1120-1127
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• 2017

Marasmius scorodonius secretes an extracellular laccase in potato dextrose broth, and this enzyme was purified up to 206-fold using $(NH_4)_2SO_4$ precipitation and a Hi-trap Q Sepharose column. The molecular mass of the purified laccase was estimated to be ~67 kDa by SDS-PAGE. The UV/vis spectrum of the enzyme was nontypical for laccases, and metal content analysis revealed that the enzyme contains 1 mole of Fe and Zn and 2 moles of Cu per mole of protein. The optimal pH for the enzymatic activity was 3.4, 4.0, and 4.6 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol, and 2,6-dimethoxy phenol as the substrate, respectively. The optimal temperature of the enzyme was $75^{\circ}C$ with ABTS as the substrate. The enzyme was stable in the presence of some metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Ni^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Ba^{2+}$, $Co^{2+}$, and $Zn^{2+}$ at a low concentration (1 mM), whereas $Fe^{2+}$ completely inhibited the enzymatic activity. The enzymatic reaction was strongly inhibited by metal chelators and thiol compounds except for EDTA. This enzyme directly decolorized Congo red, Malachite green, Crystal violet, and Methylene green dyes at various decolorization rates of 63-90%. In the presence of 1-hydroxybenzotriazole as a redox mediator, the decolorization of Reactive orange 16 and Remazol brilliant blue R was also achieved.

• 대한수의학회지
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• 제37권1호
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• pp.119-128
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• 1997

The relaxation of gastric fundus smooth muscles is the primary physiological event which induces the receptive relaxation of monogastric animals. L-arginine/Nitric oxide(L-arg/NO) system is known to mediate the inhibitory non-adrenergic non-cholinergic(NANC) neurotransmission in various tissues including gastrointestinal smooth muscles. The longitudinal smooth muscles of porcine gastric fundus showed fast relaxation during electrical field stimulation(EFS) and rebound contraction after EFS in NANC condition. So, the purpose of present study was elucidation of the neurotrasmitters related to the NANC relaxation and explanation of the relation between NANC relaxation and L-arg/NO system. The longitdinal smooth muscles of porcine gastric fundus were hung in the organ bath and under the presence of guanethidine($5{\times}10^{-5}M$), precontraction was induced by carbachol($1{\times}10^{-6}M$). The muscle responses to EFS and drugs were isomerically recorded. The rusults were summarized as follows. 1. The longtudinal muscles of porcine gastric fundus showed frequency-dependent relaxation and rebound contraction to electrical field stimulaton(1ms, 8V, 1~16Hz, 20sec, EFS). These responses were blocked by tetrodotoxin($1{\times}10^{-6}M$). 2. The relaxation and rebound contraction of the longitudinal muscles of porcine gastric fundus to EFS were inhibited by L-NAME($2{\times}10^{-5}M$). The inhibitory effect of L-NAME was antagonized by L-arginine($1{\times}10^{-3}M$), but not by D-arginine($1{\times}10^{-3}M$). 3. Exogenous NO($NaNO_2$, $1{\times}10^{-5}{\sim}1{\times}10^{-4}M$, pH=2.0) caused concentration-dependent relaxation as EFS did. 4. Methylene Blue($2{\times}10^{-5}M$), a soluble guanylate cyclase inhibitor, inhibited the relaxation and rebound contraction of the longitudinal muscles of porcine gastric fundus induced by EFS, but N-ethlmaleimide, a adenylate cyclase inhibitor, did not. 5. 8-Br-cGMP($1{\times}10^{-6}{\sim}3{\times}10^{-6}M$), permeable cGMP analogue, induced dose-dependent relaxation. but 8-Br-cAMP($1{\times}10^{-6}{\sim}3{\times}10^{-6}M$), permeable cAMP analogue, did not. Both did not evoked rebound contraction. 6. ${\alpha}$-chymotrypsin did not affect the relaxation of the longitudinal muscles of porcine gastric fundus. 7. Reactive blue 2($1{\times}10^{-4}M$, 40min) siginificantly inhibited the rebound contraction induced by EFS and inhibited contraction caused by exogenous ATP($1{\times}10^{-4}{\sim}1{\times}10^{-3}M$). These results suggests that NANC relaxation of the longitudinal muscles of porcine gastric fundus mainly mediated by NO and the rebound contraction is related to NO and other neurotransmitters.

• 대한수의학회지
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• 제38권4호
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• pp.730-736
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• 1998

To elucidate whether or not innervation of purinergic nerve and functional action of $P_{2X}$-, $P_{2Y}$-purinoceptor, the neurogenic effects of perivascular nerve stimulation were investigated using by physiograph recording system in isolated coronary artery of pig. 1. The contractile responses induced by perivascular nerve stimulation (20V, 0.5msec, 20sec) were the frequency (2~64Hz) dependent manner. 2. The neurogenic contractile responses induced by perivascular nerve stimulation were inhibited by the pretreatment with either ATP or adenosine ($10^{-7}{\sim}10^{-4}M$). 3. The neurogenic contractile responses induced by perivascular nerve stimulation (20V, 16Hz, 0.5msec, 20sec) were increased by the pretreatment with reactive blue 2, but were not affected by the pretreatment with 8-phenyltheopylline ($10^{-5}M$). 4. The neurogenic contractile responses induced by perivascular nerve stimulation (20V, 16Hz, 0.5msec, 20sec) were inhibited by the desensitization of the P2X-purinoceptor using by treatment of $10^{-5}M$, -methylene ATP as 3 times over again. The accomplished present study on isolated coronary artery of pig suggest that purinergic nerve is innervated and that the neurogenic contractile response was mediated by activation of $P_{2X}$-purinoceptor and the neurogenic relaxative response was mediated by activation of both $P_1$ and $P_{2Y}$-purinoceptor.

• The Korean Journal of Physiology and Pharmacology
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• 제14권5호
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• pp.311-316
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• 2010

It is well-known that electrical field stimulation (EFS)-induced contraction is mediated by a cholinergic mechanism and other neurotransmitters. NO, ATP, calcitonin gene-related peptide (CGRP), and substance P are released by EFS. To investigate the purinergic mechanism involved in the EFS-induced contraction, purinegic receptors antagonists were used. Suramine, a non-selective P2 receptor antagonist, reduced the contraction induced by EFS. NF023 ($10^{-7}{\sim}10^{-4}M$), a selective P2X antagonist, inhibited the contraction evoked by EFS. Reactive blue ($10^{-6}{\sim}10^{-4}M$), selective P2Y antagonist, also blocked the contraction in a dose-dependent manner. In addition, P2X agonist ${\alpha}$,${\beta}$-methylene 5'-adenosine triphosphate (${\alpha}{\beta}MeATP$, $10^{-7}{\sim}10^{-5}M$) potentiated EFS-induced contraction in a dose-dependent manner. P2Y agonist adenosine 5'-[${\beta}$-thio]diphosphate trilithium salt ($ADP{\beta}S$, $10^{-7}{\sim}10^{-5}M$) also potentiated EFS-induced contractions in a dose-dependent manner. Ecto-ATPase activator apyrase (5 and 10 U/ml) reduced EFS-induced contractions. Inversely, 6-N,$N$-diethyl-D-${\beta}$,${\gamma}$- dibromomethylene 5'-triphosphate triammonium (ARL 67156, $10^{-4}M$) increased EFS-induced contraction. These data suggest that endogenous ATP plays a role in EFS-induced contractions which are mediated through both P2X-receptors and P2Y-receptors stimulation in cat esophageal smooth muscle.

• Journal of Microbiology and Biotechnology
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• 제24권4호
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• pp.545-555
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• 2014

It has been previously demonstrated that laccases exhibit great potential for use in several industrial and environmental applications. In this paper, two laccase isoenzyme genes, lccB and lccC, were cloned and expressed in Pichia pastoris GS115. The sequence analysis indicated that the lccB and lccC genes consisted of 1,563 and 1,584 bp, and their open reading frames encoded 520 and 527 amino acids, respectively. They had 72.7% degree of identity in nucleotides and 86.7% in amino acids. The expression levels of LccB and LccC were up to 32,479 and 34,231 U/l, respectively. The recombinant laccases were purified by ultrafiltration and $(NH_4)_2SO_4$ precipitation, showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimal pH and temperature for LccB were 2.0 and $55^{\circ}C$ with 2,2'-azinobis-[ 3-ethylbenzthiazolinesulfonic acid (ABTS) as a substrate, whereas LccC exhibited optimal pH and temperature at 3.0 and $60^{\circ}C$. The apparent kinetic parameters of LccB were 0.43 mM for ABTS with a $V_{max}$ value of 51.28 U/mg, and the Km and $V_{max}$ values for LccC were 0.29 mM and 62.89 U/mg. The recombinant laccases were able to decolorize five types of dyes. Acid Violet 43 (100 g/ml) was completely decolorized by LccB or LccC (2 U/ml), and the decolorization of Reactive Blue KN-R (100 g/ml) was 91.6% by LccC (2 U/ml). Thus, the study characterizes useful laccase isoenzymes from T. versicolor that have the capability of being incorporated into the treatment of similar azo and anthraquinone dyes from dyeing industries.

• Nutrition Research and Practice
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• 제10권4호
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• pp.418-423
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• 2016

BACKGROUND/OBJECTIVES: Spirulina, a blue-green alga, is widely produced and commercialized as a dietary supplement with bio- and immune-modulatory functions. We have previously shown that spirulina had favorable effects on lipid profiles, immune functions, and antioxidant capacity in healthy Korean elderly. Despite favorable effect of spirulina supplementation, some sub-populations have shown a poor response to supplementation. Obesity is a factor related to poor-response. Therefore, the purpose of this study was to determine the immuno-modulation, antioxidant capacity, and lipid-lowering effect of spirulina in obese and non-obese Korean elderly. SUBJECTS/METHODS: The subjects were 78 elderly aged 60-87 years. In a randomized double blind, placebo-controlled study, subjects were fed either placebo or spirulina daily, at 8 g for 12 weeks. Subjects were divided into the non-obese group and the obese group based on body mass index (BMI) criteria for Asians suggested by the International Obesity Task Force: $BMI<25kg/m^2$ (non-obese) and $BMI{\geq}25kg/m^2$ (obese). RESULTS: In the non-obese group, spirulina supplementation showed a significant lowering effect on plasma concentration of total cholesterol and LDL-cholesterol, a significant increase in interleukin (IL)-2 concentration (P < 0.01) and a significant increment (P < 0.05) in IL-2/IL-6 ratio, and a significant increase in total antioxidant status level and a significant decrease in thiobarbituric acid reactive substances level. However, these effects were not observed in the obese group. CONCLUSION: These results demonstrated that blood lipid lowering and immune and antioxidant improving response for spirulina supplement was affected by obesity in Korean elderly.

• Natural Product Sciences
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• 제9권4호
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• pp.249-255
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• 2003

Zizypus is one of the herbs widely used in Korea and China due to CNS calming effect. The present study aims to investigate the effect of the methanol extract of Zizyphi Jujube Semen (ZJS) on kainic acid (KA)-induced neurotoxicity in cultured rat cerebellar granule neuron. ZJS, over a concentration range of 0.05 to $5\;{\mu]g/ml$, inhibited KA $(500\;{\mu}M)-induced$ neuronal cell death, which was measured by a trypan blue exclusion test and a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. Pretreatment of ZJS $(0.5\;{\mu}g/ml)$ inhibited KA$(50\;{\mu}M)$-induced elevation of cytosolic calcium concentration $([Ca^{2+}]_c)$, which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). ZJS $(0.5\;{\mu}g/ml)$ inhibited glutamate release into medium induced by KA $(500\;{\mu}M)$, which was measured by HPLC. These results suggest that ZJS prevents KA-induced neuronal cell damage in vitro.