• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1670-1674
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• 2007

Duffy binding protein (DBP) plays a critical role in Plasmodium vivax invasion of human red blood cells. We previously reported a single-chain antibody fragment (scFv) that was specific to P. vivax DBP (PvDBP). However, the stabilization and the half-life of scFvs have not been studied. Here, we investigated the effect of PEGylated scFvs on their biological activity and stability in vitro. SDS-PAGE analysis showed that three clones (SFDBII-12, -58, and -92) were formed as monomers (about 70 kDa) with PEGylation. Clone SFDBII-58 gave the highest yield of PEGylated scFv. Binding analysis using BIAcore between DBP and scFv showed that both SFDBII-12 and -58 were decreased approximately by two folds at the level of binding affinity to DBP after PEGylation. However, the SFDBII-92 clone still showed a relatively high level of binding affinity ($K_D=1.02{\times}10^{-7}\;M$). Binding inhibition assay showed that PEGylated scFv was still able to competitively bind the PvDBP and playa critical role in inhibiting the interactions between PvDBP protein expressed on the surface of Cos-7 cells and Duffy receptor on the surface of erythrocytes. When both scFvs and their PEGylated counterparts were exposed to trypsin, scFv was completely degraded only after 24 h, whereas 35% of PEGylated scFvs remained intact, maintaining their stability against the proteolytic attack of trypsin until 72 h. Taken together, these results suggest that the PEGylated scFvs retain their stability against proteolytic enzymes in vivo, with no significant loss in their binding affinity to target antigen, DBP.

• Nutrition Research and Practice
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• v.1 no.4
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• pp.298-304
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• 2007

This study was conducted to investigate the hypocholesterolemic effect of simvastatin (30 mg/kg BW) and antioxidant effect of coenzyme Q10 (CoQ10, 15 mg/kg BW) or green tea (5%) on erythrocyte Na leak, platelet aggregation and TBARS production in hypercholesterolemic rats treated with statin. Food efficiency ratio (FER, ADG/ADFI) was decreased in statin group and increased in green tea group, and the difference between these two groups was significant (p<0.05). Plasma total cholesterol was somewhat increased in all groups with statin compared with control. Plasma triglyceride was decreased in statin group and increased in groups of CoQ10 and green tea, and the difference between groups of statin and green tea was significant (p<0.05). Liver total cholesterol was not different between the control and statin group, but was significantly decreased in the group with green tea compared with other groups (p<0.05). Liver triglyceride was decreased in groups of statin and green tea compared with the control, and the difference between groups of the control and green tea was significant (p<0.05). Platelet aggregation of both the initial slope and the maximum was not significantly different, but the group with green tea tended to be higher in initial slope and lower in the maximum. Intracellular Na of group with green tea was significantly higher than the control or statin group (p<0.05). Na leak in intact cells was significantly decreased in the statin group compared with the control (p<0.05). Na leak in AAPH treated cells was also significantly reduced in the statin group compared with groups of the control and CoQ10 (p<0.05). TBARS production in platelet rich plasma was significantly decreased in the groups with CoQ10 and green tea compared with the control and statin groups (p<0.05). TBARS of liver was significantly decreased in the group with green tea compared with the statin group (p<0.05). In the present study, even a high dose of statin did not show a cholesterol lowering effect, therefore depletion of CoQ10 following statin treatment in rats is not clear. More clinical studies are needed for therapeutic use of CoQ10 as an antioxidant in prevention of degenerative diseases independent of statin therapy.

• BMB Reports
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• v.41 no.4
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• pp.287-293
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• 2008

In a yeast two-hybrid screen, we identified the microtubule-destabilizing protein SCG10 as a potential effector protein of $BRI_3$. The association was verified using GST pull-down, Co-IP, and their perinuclear co-localization. The analysis of in vitro microtubule polymerization/depolymerization showed that the binding of $BRI_3$ to SCG10 effectively blocked the ability of SCG10 to induce microtubule disassembly, as determined by turbidimetric assays. In intact PC12 cells, $BRI_3$ exhibited the ability to stabilize the microtubule network and attenuate the microtubule-destabilizing activity of SCG10. Furthermore, co-expression of $BRI_3$ with SCG10 attenuated SCG10-mediated PC12 cell neurite outgrowth induced by NGF. These results identify a novel connection between a neuron-specific BRI protein and the cytoskeletal network, suggesting possible roles of BRI3 in the process of neuronal differentiation.

• Applied Microscopy
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• v.35 no.4
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• pp.24-31
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• 2005

The preservative condition and ultrastructure on the mummified hair collected from newly found female mummy in Paju, were investigated by using scanning and transmission electron microscopes. The female mummy was found in september, 2002 during the traditional reburial process for the buried ones. The hair of 16th century mummy showed very intact appearances during observation with electron microscope. And the structures of the cortex, medulla and cuticle were well preserved. The cuticle layer was easily discernable, which are composed of six to seven cuticular cells. Each cuticular cells surrounded and thus seperated from its neighbors by intercellular membrane complex. In the cortex, many macrofibrils and some melanin granules between them were observed. We observed well preserved rod form macrofibrils running parallel along the direction of hair shaft. Especially, melanin granules were aggregated in the cortex which was adjacent to the cuticlu layer. As to the cause for the well-preservation of 440 year old hair sample, the presence of surface coat on the hair, which are composed of various materials. As calcium was included in the surface coat in Electron Dispersive X-ray Spectroscopy (EDS), the hardening process of the surface coat by calcium might inhibit the water or microorganism infiltration into the hair.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3333-3338
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• 2015

Background: This study was conducted to analyze positron emission tomography (PET) / computed tomography (CT) and magnetic resonance imaging (MRI) performance with oropharyngeal non-Hodgkin's lymphoma (ONHL).Materials and Methods: The complete image data of 30 ONHL cases were analyzed, all patients were performed PET / CT and MRI examination before the treatment, with the time interval of these two inspections not exceeding 14 days. The distribution, morphology, MRI signal characteristics, enhancement feature, standardized uptake value (SUV) max value and lymph node metastasis way of the lesions were analyzed. Results: Among the 30 cases, 23 cases were derived from the B-cell (76.7%), 5 cases were derived from the peripheral T cells (16.7%) and 2 cases were derived from the NK/T cells (6.7%). 19 cases exhibited the palatine tonsil involvement (63.3%). As for the lesion appearance, 10 cases appeared as mass, 8 cases were the diffused type and 12 cases were the mixed type. 25 cases exhibited the SUVmax value of PET / CT primary lesions as 11 or more (83.3%). MRI showed that all patients exhibited various degrees of parapharyngeal side-compressed narrowing, but MRI still exhibited the high-signal fat, and the oropharyngeal mucosa was intact. 25 cases were associated with the neck lymph node metastasis, among who 22 cases had no necrosis in the metastatic lymph nodes, while the rest 3 cases exhibited the central necrosis in the metastatic lymph nodes. Conclusions: PET / CT and MRI have important value in diagnosing and determining the lesion extent of ONHL.

• Clinical and Experimental Pediatrics
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• v.52 no.5
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• pp.622-626
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• 2009

Kikuchi-Fujimoto disease was initially described as a self-limiting histiocytic necrotizing lymphadenitis in Japan in 1972, and is predominantly observed in women under the age of 30 year and in Asian populations. The pathogenesis is still poorly understood but is thought to include infections, and autoimmune and neoplastic diseases. The most common clinical manifestations are fever and painless cervical lymphadenitis. Diagnosis is based on the histopathological findings, characterized by focal necrosis in the paracortical region with abundant karyorrhexis, aggregates of atypical mononuclear cells around the zone of necrosis, absence of neutrophils and plasma cells, and usually intact lymph node capsule. There is no specific therapy for the condition, and aseptic meningitis can occur as one of the complications. Here, we report the case of a patient with Kikuchi-Fujimoto disease accompanied with aseptic meningitis, which may be confused as a case of tuberculous meningitis and lymphadenitis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.3
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• pp.401-406
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• 2000

In development of the processed food, it is important not only to make the food delicious but to enhance its storage span and thermal stability without change of the food quality in color, which greatly affects the tastes of customers. Protopectinase (PPase) from Bacillus subtilis EK11 hydrolyses or dissolves protopectin in the middle lamella of plant tissues with the resultant separation of plant cells from each other, called enzymatic maceration. With the PPase, Kiwifruit was enzymatically macerated to separate cells to primary cell wall without damage. Yields of kiwifruit treated with PPase and mechanical maceration were 82% and 60%, respectively. Total and reducing sugars, crude protein and fat in the enzymatic maceration were well preserved as in the mechanical maceration. Importantly, over 95% of vitamin C, which is the most unstable component in application of the mechanical maceration, remained with intact form for one day after the enzymatic treatment. When the suspensions of kiwifruit macerated with both treatments had been stored at $4^{\circ}C for 6 days, the suspension of kiwifruit mechanically macerated was decolorized. whereas decolorization was not found in the enzymatically macerated kiwifruit. Moreover, the mechanically macerated kiwifruit was greatly deteriorated after heat treatment at $100^{\circ}C for 60 min ; the cell suspension of the exzymatically separated kiwifruit appeared to be stable, indicating the thermal stability. Thus, the PPase treatment could be a better choice for preparation of the highly valuable and functional processed food of kiwifruit as well as for prolonging the preservation period of the processed kiwifruit.

• Journal of Veterinary Clinics
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• v.26 no.5
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• pp.408-412
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• 2009

The Artemisia capillaris THUNB is a perennial herb that belongs to the family Compositae spp. and probably the most common plant among the various herbal folk remedies being used in the treatment of abdominal pain, hepatitis, chronic liver disease, jaundice and coughing in Korea. This experiment was conducted to investigate the effects of Artemisia capillaris extracts on the amounts of splenocytes-derived cytokine ($TNF-{\alpha},\;IL-1{\beta}$ and IL-10). In in vivo experimental tests using 210 ICR mice with Hepa-1c1c7 or sarcoma 180 cancer line, splenocytes derived cytokine contents were significantly (p < 0.05) reduced in the Hepa-1c1c7 and Sarcoma 180 inoculated vehicle controls, HP and SP, compared to those of the intact vehicle control on both the $28^{th}$ day and the $42^{nd}$ day, respectively. However, these decreases of $TNF-{\alpha},\;IL-1{\beta}$ and IL-10 levels induced by tumor inoculations were significantly (p < 0.01, p < 0.05) inhibited by mACH (Artemisia capillaris methanol extracts) treatment regardless of the type of experiments and tumor cells inoculated. The results suggest that Artemisia capillaris methanol extracts have prominent anti-inflammation effects on the cancer cell lines Hepa-1c1c7 and Sarcoma 180.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.4
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• pp.237-243
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• 2002

Objective : The aim of this study was to evaluate the influence of different media on blastulation, mean cell number, percentage of inner cell mass (ICM) of total cells and ICM : trophectoderm (TE) ratio in mice. Materials and methods: A total 552 two cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection (mated just after hCG injection) and cultured in MEM (n=276) or TCM (n=276) supplemented with 20% hFF. The grading of blastocysts from zona-intact (ZiB) to -escape (hatching and hatched, ZeB) was performed at 72 hours after culture. Total, TE and ICM cell numbers were analyzed by differential staining of blastocyst. Statistical analysis was performed using t-test with SigmaPlot-2001. P-values < 0.05 were accepted as statistically significant. Results: The blastulation rate in MEM ($64.9{\pm}4.95%$) was significantly higher (p=0.0031) than that in TCM ($57.2{\pm}5.22%$). No differences were found in the number of ZiB and ZeB between MEM ($31.9{\pm}2.62$, $33.0{\pm}4.58%$), and TCM ($27.2{\pm}4.28$, $30.1{\pm}4.58%$). A total 314 blastocysts (MEM=166; TCM=148) were stained differentially. Mean cell number of blastocysts was significantly higher (p=0.0002) in TCM ($73.1{\pm}3.3$) than in MEM ($61.7{\pm}2.5$). Differential staining was successfully performed in 155 blastocysts (MEM=77; TCM=78). The percentage of ICM was significantly higher in MEM than in TCM ($20.9{\pm}1.3$ vs. $17.1{\pm}1.2%$, p=0.0281). The ICM : TE ratio was higher in TCM than in MEM (1 : $4.85{\pm}0.68$ vs. 1 : $3.78{\pm}0.78$, NS). Conclusion: These results show that MEM increase the blastocyst formation and percentage of ICM of total cells comparing with TCM in mice.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.3
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• pp.249-255
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• 2015

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that $Ca^{2+}$ may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced $[Ca^{2+}]_i$ increase with nonspecific $Ca^{2+}$ channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive $[Ca^{2+}]_i$ elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a $Ca^{2+}$ chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that $Ca^{2+}$ may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream $Ca^{2+}$ regulation of the WNK-OSR1 pathway in intact cells.