• YAKHAK HOEJI
• /
• v.41 no.2
• /
• pp.212-219
• /
• 1997

Multiple forms of extracellular phospholipase $A_2$ have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase $A_2$ activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase $A_2$ activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase $A_2$ determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar $Ca^{2+}$ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase $A_2$ activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase $A_2$ from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase $A_2$, which may neither belong to the 14KDa group II phospholipase $A_2$ family nor cytosolic phospholipase $A_2$.

• Archives of Plastic Surgery
• /
• v.32 no.1
• /
• pp.124-130
• /
• 2005

In the microsurgical era, replantation with microvascular anastomosis is considered as the most superior method in aspects of texture, color, shape in case of nose amputation. There are some reported cases of replantation in nose amputation historically, but most of them are composite graft cases rather than microvascular anastomosis. Only a few cases of successful nasal replantation with microvascular anastomosis have been reported due to the reason that the size of vessels is usually very small and identifying suitable vessels for anastomosis is difficult. Microanastomosis of artery and microanastomosis of vein are ideal in replantation, but identifying suitable veins is often difficult. Without venous anastomosis, resolving the venous congestion remains to be a problem. We can carry out arteriovenous shunt if we can find two arteries in amputee. However, the smaller the size of amputee is, the more difficult it is to find two arteries. Instead of arteriovenous shunt, we can try external venous drainage(frequently swab, pin-prick, stab incision, IV or local heparin injection, dropping, apply of heparin-soaked gauze, use of medical leech). Here, we present three cases of replantation with microscopical arterial anastomosis (one angular artery, two dorsal nasal arteries) and external venous drainage (stab incision, application of medical leech and heparin-soaked gauze) even though the size of amputee may be as small as $1.5{\times}1.0cm$. In all cases, surgical outcomes were excellent in cosmetic and functional aspects. This report describes successful replantation by microvasular anastomosis in case that suitable veins are not found.

• Journal of fish pathology
• /
• v.17 no.2
• /
• pp.131-137
• /
• 2004

In the present study, phosphatidylcholine (PC) hydrolyzing enzyme had been isolated from membrane of flounder liver. PC hydrolyzing enzyme solubilized in 1% Triton X -100 from membrane was partially purified by sequential chromatography on Heparin Sepharose CL-6B and Heparin-5PW columns. The products by membrane-bound hydrolyzing enzyme were identified as phosphatidic acid and choline, but in the presence of primary alcohol, phosphatidylethanol was produced at the expense of phosphatidic acid. These data suggest that membrane-bound enzyme may be a PC-phosphoipase D (PLD) type. The enzyme had pH optimum at below 6.0 and temperature optimum at $37^\circ{C}$. The activity of PC-PLD was dose-dependently increased by $Ca^{2+}$ but not $Mg^{2+}$. The activity of PC-PLD was stimulate by PC, PIP2 and PE.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.7 no.1
• /
• pp.1-5
• /
• 2002

A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinant E. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solid-phase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below $ 30\%$, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5-3 times to about $88\%$. Besides the intrinsic affinity of angiogenin to heparin, the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, from ca. $4.8\%$ in the conventional refolding of the untagged fusion protein to $63.6\%$. The process was highly reproducible. The refolded protein in the column eluate retained RNase bioactivity of angiogenin.

• IMMUNE NETWORK
• /
• v.11 no.2
• /
• pp.123-133
• /
• 2011

Background: Mycobacterium tuberculosis (Mtb) heparin binding hemagglutinin (HBHA) is an Ag known to evoke effective host immune responses during tuberculosis infection. However, the molecular basis of the host immune response to HBHA has not been fully characterized. In this study, we examined the molecular mechanisms by which HBHA can induce the expression of proinflammatory cytokines in macrophages. Methods: HBHA-induced mRNA and protein levels of proinflammatory cytokines were determined in bone marrow-derived macrophages (BMDMs) using RT-PCR and ELISA analysis. The roles of intracellular signaling pathways for NF-${\kappa}B$, PI3-K/Akt, and MAPKs were investigated in macrophage proinflammatory responses after stimulation with HBHA. Results: HBHA robustly activated the expression of mRNA and protein of both TNF-${\alpha}$ and IL-6, and induced phosphorylation of NF-${\kappa}B$, Akt, and MAPKs in BMDMs. Both TNF-${\alpha}$ and IL-6 production by HBHA was regulated by the NF-${\kappa}B$, PI3-K, and MAPK pathways. Furthermore, PI3-K activity was required for the HBHA-induced activation of ERK1/2 and p38 MAPK, but not JNK, pathways. Conclusion: These data suggest that mycobacterial HBHA significantly induces proinflammatory responses through crosstalk between the PI3-K and MAPK pathways in macrophages.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
• /
• v.23 no.2
• /
• pp.291-299
• /
• 1993

This study was designed to evaluate the correlationship between radiologic classifications of cysts in oral region and glycosaminoglycan analysis of cystic fluids using cellulose acetate electrophoresis. The materials for this study consisted of 37 cases-8 periapical cysts, 10 dentigerous cysts, 10 primordial cysts, 2 residual cysts, 3 incisive canal cysts, 2 post-operative maxillary cysts, 1 mucocele on maxillary sinus, & 1 unicystic ameloblastoma-diagnosed as cystic lesions radiologically. The obtained results were as follows: 1. At the stepwise discriminant analysis, four variables-low mobility material, heparin, hyaluronic acid, & dermatan sulfate-were used to define diagnostic model for the odotogenic cyst. The model produced a sensitivity of 100% and a specificity of 85%. 2. The intensities of heparin and chondroitin-4-sulfate were greater in dentigerous cyst than periapical cyst(p<0.05). The intensity of chondroitin-4-sulfate was greater in primordial cyst than in periapical cyst(p<0.05). 3. It showed no statistically significant difference in glycosaminoglycan of the cystic fluids between dentigerous cyst and primordial cyst(p>0.05). 4. On the fluids of the cysts originated from maxillary sinus, there were especially high intensities of heparin and dermatan sulfate, and low intensity of chondroitin-4-sulfate. 5. On the fluids of unicystic ameloblastoma, there were high intensity of dermatan sulfate and low intenity of chondroitin-4-sulfate.

• Korean Chemical Engineering Research
• /
• v.48 no.2
• /
• pp.178-184
• /
• 2010

The aim of this work is the fabrication of polyelectrolyte microcapsules composed of biocompatible polymers such as chitosan, heparin and alginate, to encapsulate the fluorescein isothiocyanate(FITC)-albumin, and to investigate the protein release behavior therefrom. Polyelectrolyte capsules with 4-layer structures could be prepared with biocompatible materials by oppositely charged adsorption using melamin-foramide as a template. Transmission electron microscope(TEM), scanning electron microscope(SEM) and optical microscope confirmed hollow capsule structures. Protein release before and after encapsulation was monitored with a UV-Vis spectrometer. Microcapsules have different behaviors depending on the kind of polyelectrolyte polymers, chitosan-heparin capsules or chitosan-alginate capsules. In conclusion, the polyelectrolyte multilayer shells can be switched between an open and closed state by means of tuning the pH value.

• Journal of Industrial and Engineering Chemistry
• /
• v.67
• /
• pp.244-254
• /
• 2018

Surface modified poly ${\text\tiny{L}}$-lactic acid (PLLA) samples with hydroxyapatite (HA), heparin and bone morphogenetic protein-2 (BMP-2) mediated by polydopamine (pDA) coating (PLLA/pDA/HA/Hep/BMP-2) were prepared, and their effects on the enhancements of bone formation and osseointegration were evaluated in vitro and in vivo as compared to PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. The changes in surface chemical compositions, morphologies and wettabilities were observed by X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM) and water contact angle measurements. Pre-coating of HA particles with pDA provided uniform and homogeneous anchoring of particles to PLLA surface. In addition, the strong ionic interaction between heparin and pDA led PLLA surface readily heparinized for loading of BMP-2. In vitro experiments revealed that the levels of alkaline phosphatase (ALP) activity, calcium deposition, and osteocalcin (OCN) gene expression were higher in MG-63 human osteosarcoma cell lines grown on PLLA/pDA/HA/Hep/BMP-2 than on control PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. In vivo studies using micro-computed tomography (micro-CT) also showed that PLLA/pDA/HA/Hep/BMP-2 screw exhibited greatest value of bone volume (BV) and bone volume/tissue volume (BV/TV) among samples. Histological evaluations with H&E and Von Kossa staining demonstrated that a combination of HA and BMP-2 contributed to the strong osseointegration.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.25 no.3
• /
• pp.180-193
• /
• 2022

Pediatric inflammatory bowel disease (PIBD) is a multisystem disorder characterized by intestinal and extraintestinal manifestations and complications. Cerebrovascular events (CVE) are rare extraintestinal complications in patients with PIBD. Statistics show that 3.3% patients with PIBD and 1.3-6.4% adult patients with inflammatory bowel disease (IBD) experience CVE during the course of the disease. Therefore, this study aimed to review the records of children with IBD who developed CVE during the course of the disease. We retrospectively reviewed 62 cases of PIBD complicated by CVE. The mean patient age at the time of thrombotic events was 12.48±4.13 years. The incidence of ulcerative colitis was significantly higher than that of Crohn's disease (43 [70.5%] vs. 13 [21.3%] patients). Most patients (87.93%) were in the active phase of IBD at the time of CVE. The mean time interval between the onset of IBD and CVE was 20.84 weeks. Overall, 11 (26.83%) patients showed neurological symptoms of CVE at disease onset. The most frequent symptom on admission was persistent and severe headaches (67.85%). The most common site of cerebral venous thrombosis was the transverse sinuses (n=23, 53.48%). The right middle cerebral artery (n=3, 33.34%) was the predominant site of cerebral arterial infarction. Overall, 41 (69.49%) patients who were mostly administered unfractionated heparin or low-molecular-weight heparin (56.09%) recovered completely. Patients with IBD are at a risk of thromboembolism. CVE may be the most common type of thromboembolism. Based on these findings, the most common risk factor for CVE is IBD flares. In patients with CVE, anticoagulant therapy with heparin, followed by warfarin, is necessary.

• Journal of Digestive Cancer Research
• /
• v.1 no.2
• /
• pp.108-110
• /
• 2013

The relationship between malignancy and venous thromboembolism(VTE) has been well established. About 20% of all VTE cases are associated with cancer and thrombotic events are the second leading cause of death in cancer patients after death from cancer itself. Effective prophylaxis and treatment will reduce morbidity and may decrease overall mortality. We report a case of VTE in a patient with advanced gastric cancer who treated with low-molecular weighted heparin (LMWH). A 49-year-old man with heartburn was admitted to our hospital. On the endoscopic and radiologic imaging, the patient was diagnosed as an advanced gastric cancer with perigastric infiltration and liver metastasis. During the combination chemotherapy, he had pain and swelling of left lower leg. Doppler ultrasonography showed left posterior tibial venous thrombosis and pulmonary embolism CT showed thromboembolism in subsegmental pulmonary artery branch in right lower lobe. He was treated with LMWH, Dalteparin once daily via subcutaneous injection, and his symptoms was subsided.