• The Journal of Korean Physical Therapy
• /
• v.17 no.3
• /
• pp.369-376
• /
• 2005

The purpose of the study was for investigating the effect of therapeutic ultrasound irradiation on HSP70 expression in knee of degraded rat articular cartilage. Knee of ten Sprague-Dawley male rats were immobilized for 4 weeks and divided at random into the control and continuous ultrasound applicated group. The continuous ultrasound applicated group was irradiated with frequency 1MHz, intensity $1W/cm^2$ for 5 minutes. The control group was sham sonication. The immunoreactivity of HSP70 was increased in degraded articular cartilage after untrasound irradiation. These results suggest that therapeutic ultrasound can enhance HSP70 expression in degraded articular cartilage.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.30 no.5
• /
• pp.1000-1003
• /
• 2001

A yeast hydrolysate SCP-20 with anti-stress and mitigative effects for premenstrual syndromes (PMS) was composed with crude protein (57.5%) and carbohydrates (28.1%). For the investigation of anti-stress effect in the immobilization stressed rats, the weights of spleen, thyroid and kidney in the group of SCP-20 sdministration were significantly different from those in the group not given SCP-20. Total power (TP), suppression indicator of stress, was obtained from frequency of heart rate variability. TP was changed by the administration of SCP-20 suggesting that SCP-20 has the autonomous nervous control effect. In premenstrual assessment form(PAF), the administration of SCP-20 reduced the intensity of somatic symptoms, emotional symptoms, and behavioral symptoms.

• KSBB Journal
• /
• v.21 no.4
• /
• pp.279-285
• /
• 2006

We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.

• Childhood Kidney Diseases
• /
• v.3 no.1
• /
• pp.20-26
• /
• 1999

Purpose : Several modulatory factors for renal peripheral benzodiazepine receptor (PBR) has been reported, but their physiological significance remains elusive. Tissue-specific, stress-induced down-regulation of renal PBR coupled with the pharmacological stimulation of these effects by angiotensin II suggested that physiological significance of renal PBR may be related to the pathophysiology of stress-induced hypertension. The boderline hypertensive rat (BHR) has been used extensively to study the interaction of environmental factors, such as stress and blood pressure. The BHR is the first-generation progeny of a cross between the spontaneously hypertensive rat and the control Wistar-Kyoto rat. The pathogenesis of stress induced hypertension in this model is not demonstrated well. Methods In this study, BHR (male, 150-200 g) and Sprague-Dawley (SD, male, 150-200 g) rats were treated by repeated immobilization to induce anxiety. We used plus-maze performance to observe the level of anxiety by measuring percent open crosses and percent time in open. Results : Percent open crosses and percent time in open in BHR were lower than in SD rats (P<0.05). Receptor densities of renal PBR in BHRs were significantly lower than those of SDs (P<0.05). We also observed that the renal PBR was upregulated in the repeatedly stressed (immobilization, 2 hours daily, for 2 weeks) rats, both in the BHR and SD. However, the density of renal PBR in the stressed BHR was still lower than that of stressed SD. Renal PBR has been suggested to be an important organs which Is responsible for the production of cholesterol-derived products during stress. Conrlusion : From these results, it can be summarized that the lowed density of renal PBR may be involved in the pathogeneis of stress-induced hypertension.

• Korean Chemical Engineering Research
• /
• v.44 no.1
• /
• pp.1-9
• /
• 2006

Biodevices composed of biomolecular layer by mimicking the natural functions of cells and the interaction mechanisms of the constituted biomolecules have been developed in various industrial fields such as medical diagnosis, drug screening, electronic device, bioprocess, and environmental pollution detection. To construct biodevices such as bioelectronic devices (biomolecular diode, bio-information storage device and bioelectroluminescence device), protein chip, DNA chip, and cell chip, biomolecules including DNA, protein, and cells have been used. Fusion technology consisting of immobilization technology of biomolecules, micro/nano-scale patterning, detection technology, and MEMs technology has been used to construct the biodevices. Recently, nanotechnology has been applied to construct nano-biodevices. In this paper, the current technology status of biodevice including its fabrication technology and applications is described and the future development direction is proposed.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.5
• /
• pp.783-790
• /
• 2007

The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prismtype dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

• BMB Reports
• /
• v.36 no.3
• /
• pp.299-304
• /
• 2003

For deciphering the cyclic guanosine monophosphate (cGMP) signaling pathway, we employed chemical proteomics to identify the novel target molecules of cGMP. We used cGMP that was immobilized onto agarose beads with linkers directed at three different positions of cGMP. We performed a pull-down assay using the beads as baits on tissue lysates and identified 9 proteins by MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight) mass spectrometry. Some of the identified proteins were previously known cGMP targets, including cGMP-dependent protein kinase and cGMP-stimulated phosphodiesterase. Surprisingly, some of the co-precipitated proteins were never formerly reported to associate with the cGMP signaling pathway. The competition binding assays showed that the interactions are not by nonspecific binding to either the linker or bead itself, but by specific binding to cGMP. Furthermore, we observed that the interactions are highly specific to cGMP against other nucleotides, such as cyclic adenosine monophosphate (cAMP) and 5'-GMP, which are structurally similar to cGMP. As one of the identified targets, MAPK1 was confirmed by immunoblotting with an anti-MAPK1 antibody. For further proof, we observed that the membrane-permeable cGMP (8-bromo cyclic GMP) stimulated mitogen-activated protein kinase 1 signaling in the treated cells. Our present study suggests that chemical proteomics can be a very useful and powerful technique for identifying the target proteins of small bioactive molecules.

• Tuberculosis and Respiratory Diseases
• /
• v.68 no.3
• /
• pp.140-145
• /
• 2010

Background: Pulmonary embolism (PE) is a common clinical problem in the West that is associated with substantial morbidity and mortality. The diagnostic modality has been changed since 2001. This study retrospectively reviewed the PE mortality with the aim of identifying the risk factors associated with mortality since the multidetector computed tomography (MDCT) was introduced. Methods: We analyzed 105 patients with acute PE proven by multidetector CT or ventilation perfusion scan. The primary outcome measure was the all-cause mortality at 3 months. The prognostic effect of the baseline factors on survival was assessed by multivariate analysis. Results: The main risk factors were prolonged immobilization, stroke, cancer and obesity. Forty nine percent of patients had 3 or more risk factors. The overall mortality at 3 months was 18.1%. Multivariate analysis revealed low diastolic blood pressure and the existence of cancer to be independent factors significantly associated with mortality. Forty two PE patients were examined for the coagulation inhibitors. Four of these patients had a protein C deficiency (9.5%), and 11 had a protein S deficiency (26%). Conclusion: PE is an important clinical problem with a high mortality rate. Close monitoring may be necessary in patients with the risk factors.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.11
• /
• pp.2037-2043
• /
• 2017

The surface protein hemagglutinin (HA) mediates the attachment of influenza virus to host cells containing sialic acid and thus facilitates viral infection. Therefore, HA is considered as a good target for the development of diagnostic tools for influenza virus. Previously, we reported the isolation of single-stranded aptamers that can distinguish influenza subtype H1 from H5. In this study, we describe a method for the selective electrical detection of H1 using the isolated aptamer as a molecular probe. After immobilization of the aptamer on Si wafer, enzyme-linked immunosorbent assay (ELISA) and field emission scanning electron microscopy (FE-SEM) showed that the immobilized aptamer bound specifically to the H1 subtype but not to the H5 subtype. Assessment by cyclic voltammetry (CV) also demonstrated that the immobilized aptamer on the indium thin oxide-coated surface was specifically bound to the H1 subtype only, which was consistent with the ELISA and FE-SEM results. Further measurement of CV using various amounts of H1 subtype provided the detection limit of the immobilized aptamer, which showed that a nanomolar scale of target protein was sufficient to produce the signal. These results indicated that the selected aptamer can be an effective probe for distinguishing the subtypes of influenza viruses by monitoring current changes.

• Applied Chemistry for Engineering
• /
• v.1 no.2
• /
• pp.107-115
• /
• 1990

New insulin-releasing system on the basis of the redox reaction of glucose was synthesized by immobilizing insulin through a disulfide bond(5, 5'-dithiobis(2-nitrobenzoic acid) to polymer membrane(poly(methyl methacrylate)) and enzyme(glucose oxidase). The disulfide bonds were cleaved upon oxidation of glucose with glucose dehydrogenase and glucose oxidase, releasing insulin from the membrane and enzyme. Sensitivity to glucose concentration was enhanced by coimmobilization of enzyme cofactors(nicotinamide adenin dinucleotide and flavin adenin dinucleotide) acting as electron mediator(for the membrane device), and further enhanced by direct immobilization of insulin on glucose oxidase(for the protein device). Both systems were specific to glucose, and the released insulin was indistinguishable from native insulin. The biological activity of released insulin was 81% of native insulin.