• The Korean Journal of Physiology
• /
• 제22권2호
• /
• pp.245-257
• /
• 1988

The present study was undertaken to investigate the effect of clonidine on the response of the dorsal horn cells to intra-arterially administered bradykinin $(BK:40{\mu}g)$ and $K^+(4mg)$ in spinal cats and cats with intact spinal cord. The change in the activities of low threshold (LT), high threshold (HT) and wide dynamic range (WDR) cells induced by BK and $K^+$ were determined before and after treatment of animals with clonidine. Also studied was mechanism of inhibitory action of clonidine on the responses of dorsal horn cells to the chemical algogenics. Number of WDR cell responded to intra-arterially administered BK and $K^+$ was greater in spinal animals than in cats with intact spinal cord. Following administration of BK or $K^+$ no change was observed in the activity of LT cell whereas activity of HT cell increased invariably. The increased response of HT cell to BK and $K^+$ was markedly suppressed by clonidine. On the other hand, such inhibitory actions of clonidine were almost completely blocked by yohimbine. The majority of WDR cells were activated by $K^+$ while response of WDR cells to BK was diverse (excitatory, inhibitory or mixed). These results indicate that clonidine inhibits responses of the dorsal horn cells not only to thermal or mechanical stimulations but also to chemical algogenics, and that the inhibitory action of clonidine is generally mediated through excitation of ${\alpha}_2-adrenoreceptors$.

• Animal Bioscience
• /
• 제36권2호
• /
• pp.315-321
• /
• 2023

Objective: The study was conducted to investigate the dephosphorylation of Pseudomonas aeruginosa flagellin (PA FLA) by sweet potato purple acid phosphatase (PAP) and the effect of the enzyme on the flagellin-mediated inflammatory response in the A549 lung epithelial cell line. Methods: The activity of sweet potato PAP on PA FLA was assayed at different pH (4, 5.5, 7, and 7.5) and temperature (25℃, 37℃, and 55℃) conditions. The release of interleukin-8 (IL-8) and the activation of nuclear factor kappa- light-chain-enhancer of activated B cells (NF-κB) in A549 cells exposed to PA FLA treated with or without sweet potato PAP was measured using IL-8 and NF-κB ELISA kits, respectively. The activation of toll-like receptor 5 (TLR5) in TLR5-overexpressing HEK-293 cells exposed to PA FLA treated with or without sweet potato PAP was determined by the secreted alkaline phosphatase-based assay. Results: The dephosphorylation of PA FLA by sweet potato PAP was favorable at pH 4 and 5.5 and highest at 55℃. PA-FLA treated with the enzyme decreased IL-8 release from A549 cells to about 3.5-fold compared to intact PA FLA at 1,000 ng/mL of substrate. Moreover, PA-FLA dephosphorylated by the enzyme repressed the activation of NF-κB in the cells compared to intact PA FLA. The activation of TLR5 by PA-FLA was highest in TLR-overexpressing HEK293 cells at a substrate concentration of 5,000 ng/mL, whereas PA FLA treated with the enzyme strongly repressed the activation of TLR5. Conclusion: Sweet potato PAP has the potential to be a new alternative agent against the increased antibiotic resistance of P. aeruginosa and may be a new conceptual feed additive to control unwanted inflammatory responses caused by bacterial infections in animal husbandry.

• 대한세포병리학회지
• /
• 제14권1호
• /
• pp.17-21
• /
• 2003

Insular carcinoma of the thyroid(ICT) is an uncommon thyroglobulin-producing neoplasm, intermediate between well differentiated and anaplastic carcinoma. Only a few publications have addressed the fine needle aspiration cytologic(FNAC) findings from ICT. We experienced a case histologically diagnosed as ICT and with preoperative FNAC in a 52 - year - old woman. The FNAC displayed scanty colloid and abundant monomorphic follicular cells presented singly, in small loose aggregates, and in cohesive trabecular or acinar clusters. Intact insulae of tumor cells were also identified. Necrosis and mitosis were rare. Tumor cells showed round and monomorphic nuclei, finely granular chromatin, and inconspicuous nucleoli. When insular structure is identified in thyroid FNAC specimen, ICT should be included in the differential diagnosis. Herein we discuss and review the cytologic criteria for separation of ICT from other thyroid neoplasms.

• Journal of Microbiology and Biotechnology
• /
• 제19권8호
• /
• pp.836-844
• /
• 2009

A denitrification bacterium was isolated from riverbed soil and identified as Ochrobactrum sp., whose specific enzymes for denitrification metabolism were biochemically assayed or confirmed with specific coding genes. The denitrification activity of strain G3-1 was proportional to glucose/nitrate balance, which was consistent with the theoretical balance (0.5). The modified graphite felt cathode with neutral red, which functions as a solid electron mediator, enhanced the electron transfer from electrode to bacterial cell. The porous carbon anode was coated with a ceramic membrane and cellulose acetate film in order to permit the penetration of water molecules from the catholyte to the outside through anode, which functions as an air anode. A non-compartmented electrochemical bioreactor (NCEB) comprised of a solid electron mediator and an air anode was employed for cultivation of G3-1 cells. The intact G3-1 cells were immobilized in the solid electron mediator, by which denitrification activity was greatly increased at the lower glucose/nitrate balance than the theoretical balance (0.5). Metabolic stability of the intact G3-1 cells immobilized in the solid electron mediator was extended to 20 days, even at a glucose/nitrate balance of 0.1.

• Journal of Microbiology and Biotechnology
• /
• 제25권12호
• /
• pp.2034-2042
• /
• 2015

A one-pot process of enzymatic synthesis of deoxythymidine-5'-triphosphate (5'-dTTP) employing whole cells of recombinant Escherichia coli coexpressing thymidylate kinase (TMKase) and acetate kinase (ACKase) was developed. Genes tmk and ack from E. coli were cloned and inserted into pET28a(+), and then transduced into E. coli BL21 (DE3) to form recombinant strain pTA in which TMKase and ACKase were simultaneously overexpressed. It was found that the relative residual specific activities of TMKase and ACKase, in pTA pretreated with 20 mM ethylene diamine tetraacetic acid (EDTA) at 25℃ for 30 min, were 94% and 96%, respectively. The yield of 5'-dTTP reached above 94% from 5 mM deoxythymidine 5'-monophosphate (5'-dTMP) and 15 mM acetyl phosphate catalyzed with intact cells of pTA pretreated with EDTA. The process was so effective that only 0.125 mM adenosine-5'-triphosphate was sufficient to deliver the phosphate group from acetyl phosphate to dTMP and dTDP.

• KSBB Journal
• /
• 제9권2호
• /
• pp.122-126
• /
• 1994

A Aureobasidium pullulans 균체를 이용하여 malt-ose로부터 isomalto-oligosaccharides를 생산하였다. 저농도의 maltose를 기질로 이용할 경우에는 가수분해활성이 transglucosylation활성보다 매우 강하여 isomalto-oligosaccharides 합성수율이 매우 낮았으나 400g/ P 이상의 고농도기질에서는 isomalto-oligosaccharides 합성이 유리하였다. isomaltomalto-oligosaccharides 생산의 반응 최적조건은 pH 4.5, 온도 $65^{\circ}C$, 균체농도는 기질 그램당 10unit이었다. 이때 생산된 isomaltooligosa-ccharides의 수율은 최고 48.35%였다. 반응초기에 maltose로부터 panose가 선택적으로 생산되었고, maltose가 더 이상 기질로 이용되지 못하는 반응시점에서 panose를 기질로 이용하여 isomaltotriose가 생산되었다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권2호
• /
• pp.245-251
• /
• 2006

The aim of this study was to investigate the effects of caponization on the bone characteristics, biomechanical property and histology in Taiwan country chickens fed to market age of 26 wks. Male Taiwan country chickens $D{\times}L_2$ were caponized or sham-operated at 8 wks of age, and selected healthy sham-operated and completely caponized chickens (prominent degenerated comb) were selected at 16 wks old and fed to 26 wks old for the trials. Fifteen intact male chickens (Intact), sham-operated chickens (Sham) and caponized chickens (Capon) were assigned for trial 1, and sixteen Intact and Capon were assigned for trial 2. Results in trial 1 showed that the abdominal fat and relative abdominal fat weights of Capon were significantly heavier than Intact and Sham (p<0.05), while the tibia weight and relative weight were the lightest (p<0.05). The tibia breaking strength, bending moment and stress of Capon were the poorest among groups (p<0.05). The trial 2 produced the similar observation that Capon were significantly lighter than Intact (p<0.05) in the tibia weight, relative tibia weight and their biomechanical properties. On histological determinations, Capon showed a thinner cartilage end and fewer chondrocytes (about 50%) and trabecular, and bigger marrow cavity; while decreased hemopoietic cells number with increased adipocytes than Intact observed by H&E stain and at low magnification. At high magnification, Capon showed a decrease in the chondrocyte size by 33 to 50%, with smaller nucleus located near the cell membrane, and exhibited monocellular form chondrocytes. Capon also showed a less strongly acidic sulfated mucosubstance with weaker dyeing property within cartilage zone, and smaller chondrocytes size by Alcian blue stain.

• Molecules and Cells
• /
• 제26권2호
• /
• pp.113-120
• /
• 2008

Laser light microscopy enables observation of various simultaneously occurring events in living cells. This capability is important for monitoring the spatiotemporal patterns of the molecular interactions underlying such events. Two-photon excited fluorescence microscopy (two-photon microscopy), a technology based on multiphoton excitation, is one of the most promising candidates for such imaging. The advantages of two-photon microscopy have spurred wider adoption of the method, especially in neurological studies. Multicolor excitation capability, one advantage of two-photon microscopy, has enabled the quantification of spatiotemporal patterns of $[Ca^{2+}]_i$ and single episodes of fusion pore openings during exocytosis. In pancreatic acinar cells, we have successfully demonstrated the existence of "sequential compound exocytosis" for the first time, a process which has subsequently been identified in a wide variety of secretory cells including exocrine, endocrine and blood cells. Our newly developed method, the two-photon extracellular polar-tracer imaging-based quantification (TEPIQ) method, can be used for determining fusion pores and the diameters of vesicles smaller than the diffraction-limited resolution. Furthermore, two-photon microscopy has the demonstrated capability of obtaining cross-sectional images from deep layers within nearly intact tissue samples over long observation times with excellent spatial resolution. Recently, we have successfully observed a neuron located deeper than 0.9 mm from the brain cortex surface in an anesthetized mouse. This microscopy also enables the monitoring of long-term changes in neural or glial cells in a living mouse. This minireview describes both the current and anticipated capabilities of two-photon microscopy, based on a discussion of previous publications and recently obtained data.

• 한국식품과학회지
• /
• 제29권5호
• /
• pp.1021-1027
• /
• 1997

Dicyma sp. YCH-37이 생산하는 효모세포벽 용해효소의 성질을 검토한 결과, 각종 환원제와 금속이온에 대체로 안정하였고, guanidine-HCl을 제외한 여러 화학수식제에 대해서도 안정하였다. 배양시간, 전처리 및 배양조건에 따른 영향을 검토한 결과, 정지기 및 사멸기에 있는 효모보다는 대수증식기의 효모, 그리고 생효모에 비해 열처리된 효모가 더 잘 용균되었다. Butanol, acetone 등의 유기용매로 처리된 효모가 그렇지 않은 효모보다 용균도가 좋았으며, 0.5 M ammonium sulfate가 함유된 Yeast extract-Malt extract 배지에서 생육한 효모, 그리고 진탕배양한 효모보다 정치배양한 효모가 용균효소에 의해 더 잘 용균되었다. SDS, Triton X-100, ${\beta}-mercaptoethanol$, potassium chloride, sodium sulfite 등의 화학수식제를 효소반응액에 첨가하였을 때 기질 효모는 더 잘 용균되었다.

• 식물조직배양학회지
• /
• 제21권5호
• /
• pp.309-314
• /
• 1994

간편하면서도 효율적인 단자엽 식물의 형질전환 기법을 개발하기 위하여 배발생 세포를 직접 electroporation하여 DNA를 도입하는 실험을 벼와 잔디에서 실시하였다. 잔디는 수정 후 3주된 미숙배에서 캘러스를 유도하였으며, 2.4-D가 1 mg/L 함유된 액체 MS배지로 옮겨 진탕배양한 것을 electroporation 실험에 이용하였다. 벼는 20 mm 정도의 미숙화서 유래의 캘러스를 액체 N$_{6}$배지(1 mg/L 2.4-D 함유)에서 진탕배양하여 획득한 세포주를 사용하였다. 액체 진탕배양한 세포괴를 GUS expression vector인 pGA1074 (30 $\mu\textrm{g}$/ml)와 함께 MS 액체 배지에서 Electroporation하였다. 세포벽과 세포막을 통한 세포로의 DNA 전이는 GUS 유전자의 발현 여부 및 정도에 따라 결정하였다. 400 volt, 800 $\mu$F capacitance로 electroporation 처리된 벼와 잔디의 세포괴들은 200 ${\mu}\ell$ (packed cell volume)의 세포괴 당 25 unit (1 unit=파란색을 띤 독립된 세포군) 이상의 빈도로 GUS 활성을 나타내었다. 반면에 무처리 세포주 및 처리한 비배발생 세포주에서는 GUS 발현이 일어나지 않음을 반복적으로 확인차였다. 따라서 electroporation에 의한 벼와 잔디의 형질전환실험에서 원형질체 대신 intact한 배발생 세포가 이용될 수 있음을 의미한다