• The Korean Journal of Pain
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• 제21권2호
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• pp.112-118
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• 2008

Background: This study was conducted to quantitatively analyze proteins associated with the calcitonin gene-related peptide (CGRP) in cerebrospinal fluid (CSF) that was obtained from a rat model of chronic neuropathic pain following administration of intrathecal $CGRP_{8-37}$. Methods: Male Sprague-Dawley rats (100-150 g, 5-6 wks) were divided into two groups, sham controls and neuropathic pain models. At the time of operation for neuropathic pain model, an intrathecal catheter was threaded through the intrathecal space. At 1 or 2 wks after the operation (maximum pain state), a test dose of 1, 5, 10, or 50 nM of $CGRP_{8-37}$ was injected into the intrathecal catheter and the CSF was then aspirated. Conventional proteomics to evaluate the CSF were then performed using high resolution 2-D, gel electrophoresis followed by computational image analysis and protein identification by mass spectrometry. Results: Treatment with $CGRP_{8-37}$ effectively alleviated mechanical allodynia in a dose dependent manner. The most effective response was obtained when a dose of 50 nM was administered, but significant differences were obtained following administration of only 5 nM $CGRP_{8-37}$. Furthermore, the results of the proteomic analysis were consistent with the experimental results. Specially we detected 30 differentially expressed spots in 7 images when 2-D gel electrophoresis was conducted. The intensity of 6 of these spots (spot number: 20 and 26-30) was found decrease the $CGRP_{8-37}$ dose increased; therefore, these spots were evaluated by mass spectrometry. This analysis identified 2 different proteins, CGRP (spot numbers: 26-30) and neurotensin-related peptide (spot number: 20). Conclusions: The results of this study suggest that CGRP plays a role in chronic central neuropathic pain and is a major target of chronic neuropathic pain management.

• The Plant Pathology Journal
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• 제34권6호
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• pp.499-505
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• 2018

Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.

• 대한약침학회지
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• 제20권2호
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• pp.119-126
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• 2017

Objectives: Semecarpus anacardium Linn. is a plant well-known for its antimicrobial, antidiabetic and anti-arthritic properties in the Ayurvedic and Siddha system of medicine. This has prompted the screening of this plant for antibacterial activity. The main aims of this study were to isolate compounds from the plant's seeds and to evaluate their antibacterial effects on clinical bacterial test strains. Methods: The n-butanolic concentrate of the seed extract was subjected to thin layer chromatography (TLC) and repeated silica gel column chromatography followed by elution with various solvents. The compound was identified based on observed spectral (IR, $^1H$ NMR, $^{13}C$ NMR and high-resolution mass spectrometry) data. The well diffusion method was employed to evaluate the antibacterial activities of the isolated acyclic isoprenoid compound (final concentration: $5-15{\mu}g/mL$) on four test bacterial strains, namely, Staphylococcus aureus (MTCC 96), Bacillus cereus (MTCC 430), Escherichia coli (MTCC 1689) and Acinetobacter baumannii (MTCC 9829). Results: Extensive spectroscopic studies showed the structure of the isolated compound to be an acyclic isoprenoid ($C_{21}H_{32}O$). Moreover, the isoprenoid showed a remarkable inhibition of bacterial growth at a concentration of $15{\mu}g/mL$ compared to the two other doses tested (5 and $10{\mu}g/mL$) and to tetracycline, a commercially available antibiotic that was used as a reference drug. Conclusion: The isolation of an antimicrobial compound from Semecarpus anacardium seeds validates the use of this plant in the treatment of infections. The isolated compound found to be active in this study could be useful for the development of new antimicrobial drugs.

• 전자공학회논문지SC
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• 제40권3호
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• pp.172-180
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• 2003

본 연구에서는 자기공명영상(MRI)에서 수소 원자핵의 공명주파수(PRF) 방법을 기반으로 인체 종아리 근육 외부의 열원에 의해 근육 내부로 열원이 전달되는 과정을 비침습적으로 관찰하는 방법을 제시한다. 열전달과정을 온도 변화로 측정하였는데 온도 영상의 안정성 및 보정 실험은 phantom을 이용하였고 온도의 변화는 phantom과 인체 모두에서 측정하였다. Phantom 실험은 agarose gel을 중탕하여 약 50℃까지 가열시킨 후 1시간의 냉각과정 동안 매 3분마다 데이터를 획득하였다. 인체 실험에서는 지원자의 종아리(the calf)에 hot pack을 이용하여 열을 전달하였다. Hot pack을 발열시키기 전에 기준 데이터를 1번 획득하고, 발열시킨 후부터 매 2분마다 30분 동안 데이터를 획득하였다. 획득된 영상 데이터는 위상차 영상으로 재구성된 다음 각 ROI에서의 평균 위상차를 관측하였다. 온도를 34.2∼50.2℃의 범위에서 변화시켰을 때 phantom의 위상차는 온도 변화에 대해 선형적으로 변하였다. 이 범위에서 측정된 온도의 해상도는 0.0457 radian/℃(0.0038 ppm/℃)였다. 인체 실험에서는 각 영상에서 hot pack과 가까운 위치의 평균 위상차가 hot pack과 먼 위치의 평균 위상차보다 작은 값을 나타냈다 이를 통해 같은 영상 단면에서도 열원(heat source)과의 거리에 따라서 온도 변화가 다르게 나타나는 것을 관찰할 수 있었다. 본 연구를 통해 PRF방법을 이용하여 MRI에서도 비침습적으로 인체 내부의 열전달과정을 관측하였고 이로서 온열치료 시 MRI가 임상적 이용 가능성이 있음을 확인하였다.

• Journal of Microbiology
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• 제45권6호
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• pp.499-504
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• 2007

A new antagonistic strain of actinomycete, designated AP19-2, was isolated from the feces of giant pandas inhabiting the Foping National Nature Reserve in China. Cultural characteristic studies strongly suggested that this strain is a member of the genus Streptomyces. The nucleotide sequence of the 16S rRNA gene of strain AP19-2 evidenced profound similarity (97-99 %) with other Streptomyces strains. Two pure active molecules were isolated from a fermentation broth of Streptomyces sp. strain AP19-2 via extraction, concentration, silica gel G column chromatography, and HPLC. The chemical structures of the two related compounds (referred to as chromomycin $A_2$ and chromomycin $A_3$) were established on the basis of their Infrared spectra (IR), High Resolution Electrospray Ionization Mass Spectrometry (HR-ESI-MS), and Nuclear Magnetic Resonance (NMR) data, and by comparison with published data.

• 대한소아치과학회지
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• 제24권1호
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• pp.65-81
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• 1997

For the purpose of evaluating the appropriateness of AP-PCR as a facile, rapid and reproducible method for genotyping Streptococcus mutans, and selecting the discriminant primer for it, a DNA fingerprinting was performed on the microorganisms isolated from caries-free children and children with rampant caries, respectively. In the course of selecting appropriate primer for S. mutans genotyping, we chose S2 primer from 6 different primers which shows highest resolution on the agarose gel as well. Nineteen kinds of fingerprint patterns were observed in caries-free children and children with rampant caries which were produced by combination of 7 different fragments. Interestingly, the number of types observed in caries-free children was greater than that in children with rampant caries. And we observed Type 2 was predominant in children with rampant caries (about 80%) and relatively even distribution of each types in caries-free children. Furthermore, it was appeared that the major types in normal control were not or rarely found in children with rampant caries. In conclusion, we could establish simple, rapid and highly reproducible AP-PCR method for genotyping S. mutans. We also found differences in distribution of S. mutans between normal and patient, which suggested that cariogenicity is also dependent on qualitative aspects which is caused by the difference in genotypes of S. mutans in oral cavity.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권4호
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• pp.293-300
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• 2001

A polyrotaxane-biotin conjugate was synthesized and its interaction with streptavidin measured using surface plasmon resonance(SPR) detection. A biodegradable polyrotaxane in which ca, 22 molecules of ${\alpha}$-cyclodextrina(${\alpha}$-CDs) were threaded onto a poly(ethylene oxide) chain(M$\sub$n:4,000) capped with benzyloxycarbonyl-L-phenylalanine was conjugated with a biotin hydorazide and 2-aminoethanol after activing the hydroxyl groups of ${\alpha}$-CDs in the polyrotaxane using N, N'-carbonyldiimidazole. The results of the high-resolution $^1$H-nyclear lmagnetic resonance($^1$H-NMR)spectra and gel permeation chromatography of the conjugate showed that ca, 11 biotin molecules were actually introduced to the polyrotaxane scaffold. An SPR analysis showed that the binding curves of the biotin molecules in the conjugate on the streptavidin-deposited surface changed in a concentration dependent manner, indicating that the biotin in the conjugate was ac-tually recognized by streptavidin. The association equilibrium constant(K$\sub$a/) of the interaction be-tween the conjugate and steptavidin tetramer was of the order 10$\^$7/. These results suggest that polyrotaxane is useful for scaffolds as a polymeric ligand in biomedical fields.

• Food Science and Biotechnology
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• 제18권4호
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• pp.823-832
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• 2009

In recent years, special concerns have been raised about the safety assessment of foods and food ingredients derived from genetically modified organisms (GMOs). A growing number of countries establish regulations and laws for GMOs in order to allow consumers an informed choice. In this case, a lot of methods have been developed for the detection of GMOs. However, the reproducibility among methods and laboratories is still a problem. Consequently, it is still in great demand for more effective methods. In comparison with the gel electrophoresis, the capillary electrophoresis (CE) technology has some unique advantages, such as high resolution efficiency and less time consumption. Therefore, some CE-based methods have been developed for the detection of GMOs in recent years. All kinds of CE detection methods, such as ultraviolet (UV), laser induced fluorescence (LIF), and chemiluminescence (CL) detection, have been used for GMOs detection. Microchip capillary electrophoresis (MCE) methods have also been used for GMOs detection and they have shown some unique advantages.

• Journal of Microbiology and Biotechnology
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• 제16권11호
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• pp.1832-1836
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• 2006

Mesorhizobium sp. LUK, which utilizes 3-amino-3-phenylpropionic acid as the sole source of nitrogen with high enantioselectivity (E(S)>100), was isolated using enrichment culture. The enzyme involved in the utilization of (S)-3-amino-3-phenylpropionic acid was confirmed to be a transaminase and was purified by 235-folds with a specific activity of 0.72 U/mg. The molecular weight of the purified protein was ca. 47 kDa and the active enzyme was determined as a dimer on gel filtration chromatography. The N-terminal sequence was obtained from the purified protein. Spontaneous decarboxylation of produced ${\beta}-keto$ acids was observed during the chiral resolution of 3-amino-3-phenylpropionic acid.

• Journal of Nutrition and Health
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• 제29권9호
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• pp.971-980
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• 1996

The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.