• Archives of Plastic Surgery
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• v.39 no.2
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• pp.162-165
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• 2012

The synchondrosis between the sixth and seventh costal cartilage is usually used for the base frame in autogenous ear reconstruction. If the synchondrosis is loose, a variety of modifications can be devised. This report introduces new methods for these problems. In cases of incomplete synchondrosis, only the surface of the base block margin was smoothly tapered without carving for the removal of the conchal deepening. The secure fixation of the two segments (helix and antihelix) to the base block using fine wire sutures gave stability to the unstable basal frame. After confirming that all the segments were assembled in one stable piece, the remaining conchal deepening of the basal framework was removed, and the outer lower portion of the basal cartilage was trimmed along its whole length. A total of 10 consecutive patients with microtia, ranging from 8 to 13 years old, were treated from 2008 to 2009. The follow-up period was 6 months to 2 years. Despite incomplete synchondrosis, the stable frameworks were constructed using the authors' method and aesthetically acceptable results were achieved. The proposed method can provide an easy way to make a stable cartilage framework regardless of the variable conditions of synchondrosis.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1261-1271
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• 2020

Cytochrome c_L (Cytc_L) is an essential protein in the process of methanol oxidation in methylotrophs. It receives an electron from the pyrroloquinoline quinone (PQQ) cofactor of methanol dehydrogenase (MDH) to produce formaldehyde. The direct electron transfer mechanism between Cytc_L and MDH remains unknown due to the lack of structural information. To help gain a better understanding of the mechanism, we determined the first crystal structure of heme c containing Cytc_L from the aquatic methylotrophic bacterium Methylophaga aminisulfidivorans MP^T at 2.13 Å resolution. The crystal structure of Ma-Cytc_L revealed its unique features compared to those of the terrestrial homologues. Apart from Fe in heme, three additional metal ion binding sites for Na⁺, Ca⁺, and Fe²⁺ were found, wherein the ions mostly formed coordination bonds with the amino acid residues on the loop (G93-Y111) that interacts with heme. Therefore, these ions seemed to enhance the stability of heme insertion by increasing the loop's steadiness. The basic N-terminal end, together with helix α4 and loop (G126 to Y136), contributed positive charge to the region. In contrast, the acidic C-terminal end provided a negatively charged surface, yielding several electrostatic contact points with partner proteins for electron transfer. These exceptional features of Ma-Cytc_L, along with the structural information of MDH, led us to hypothesize the need for an adapter protein bridging MDH to Cytc_L within appropriate proximity for electron transfer. With this knowledge in mind, the methanol oxidation complex reconstitution in vitro could be utilized to produce metabolic intermediates at the industry level.

• Applied Biological Chemistry
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• v.51 no.4
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• pp.276-280
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• 2008

To elucidate the effects of ultraviolet (UV) irradiation on molecular properties of ovalbumin, the molecular weight profile, secondary structure and tertiary structure of proteins were examined after irradiation by UV with 254 nm wavelength for 4, 8, 16 and 32 hrs, respectively. UV irradiation of protein solution caused the disruption on the native state of protein molecules. SDS-PAGE and gel permeation chromatography indicated that radiation caused initial fragmentation of polypeptide chains and as a result subsequent aggregation due to cross-linking of protein molecules. Circular dichroism (CD) study showed that UV irradiation caused the change on the secondary structure resulting in decrease of helical structure or compact denature on structure of protein depending on irradiation period. Fluorescence spectroscopy indicated that irradiation quenched the emission intensity excited at 280 nm. These results suggest that UV irradiation affect the molecular properties of ovalbumin and may have potential as a means to change the antigenicity of protein allergen.

• BMB Reports
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• v.38 no.5
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• pp.624-631
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• 2005

Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the main diseases to mankind. It is urgent to discover novel drug targets for appropriate antimicrobial agents against this human pathogen. The shikimate pathway is onsidered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammalian cells. The Mycobacterium tuberculosis aroE-encoded shikimate dehydrogenase was cloned, expressed and purified. Sequence alignment analysis shows that shikimate dehydrogenase of Mycobacterium tuberculosis exhibit the pattern of G-X-(N/S)-V-(T/S)-X-PX-K, which is highly conserved within the shikimate dehydrogenase family. The recombinant shikimate dehydrogenase spectrum determined by CD spectroscopy showed that the percentages for $\alpha$-helix, $\beta$-sheet, $\beta$-turn, and random coil were 29.2%, 9.3%, 32.7%, and 28.8%, respectively. The enzymatic characterization demonstrates that it appears to be fully active at pH from 9.0 to 12, and temperature $63^{\circ}C$. The apparent Michaelis constant for shikimic acid and $NADP^+$ were calculated to be about $29.5\;{\mu}M$ and $63\;{\mu}M$. The recombinant shikimate dehydrogenase catalyzes the substrate in the presence of $NADP^+$ with an enzyme turnover number of $399\;s^{-1}$. Zymological studies suggest that the cloned shikimate dehydrogenase from M. tuberculosis has a pretty activity, and the work should help in the discovery of enzyme inhibitors and further of possible antimicrobial agents against Mycobacterium tuberculosis.

• BMB Reports
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• v.40 no.3
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• pp.412-418
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• 2007

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is an unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes. In the present work, the entire encoding region for Oryza sativa PHGPx was expressed in Escherichia coli M15, and the purified fusion protein showed a single band with 21.0 kD and pI = 8.5 on SDS- and IFE-PAGE, respectively. Judging from CD and fluorescence spectroscopy, this protein is considered to have a well-ordered structure with 12.2% $\alpha$-helix, 30.7%$\beta$-sheet, 18.5% $\delta$-turn, and 38.5% random coil. The optimum pH and temperature of the enzyme activity were pH 9.3 and 27$^{\circ}C$. The enzyme exhibited the highest affinity and catalytical efficiency to phospholipid hydroperoxide employing GSH or Trx as electron donor. Moreover, the protein displayed higher GSH-dependent activity towards t-Butyl-OOH and $H_2O_2$. These results show that OsPHGPx is an enzyme with broad specificity for hydroperoxide substrates and yielded significant insight into the physicochemical properties and the dynamics of OsPHGPx.

• Biomedical Science Letters
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• v.21 no.3
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• pp.152-159
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• 2015

Der p 2, which is a major allergen of house dust mite, plays an important role in the pathogenesis of allergic disease. There is controversy regarding whether Der p 2 binds to Toll-like receptor 4 (TLR4), and its inflammatory effect has not yet been elucidated. In the current study, we examined the interaction of Der p 2 with TLR4 and the effect of Der p 2 on cytokine release in THP-1 cells and lymphocytes. Among house dust mite extracts, recombinant TLR4 protein interacted with Der p 2. The overall structure of Der p 2 is characteristic of the immunoglobulin superfamily and contains ten ${\beta}-strands$, forming a ${\beta}-cup$ fold with two anti-parallel ${\beta}-sheets$, and a short 310 helix. The two sheets can be separated, further allowing the formation of a large internal pocket, which is narrow and suitable for binding large flat molecules such as lipid-like molecules. Der p 2 caused increased secretion of IL-6, IL-8, and MCP-1, which are neutrophil survival factors, in human monocytic THP-1 cells in a time-dependent manner. Der p 2 also induces the release of cytokines in normal and allergic lymphocytes. Supernatant after treatment with Der p 2 inhibited neutrophil apoptosis. In coculture of lymphocytes with neutrophils, Der p 2 inhibited spontaneous apoptosis of allergic neutrophils. In summary, Der p 2 binds to TLR4 and induces an inflammatory response such as cytokine secretion in immune cells. These findings may enable elucidation of allergy pathogenesis by specific allergen of house dust mite.

• Archives of Plastic Surgery
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• v.40 no.3
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• pp.192-197
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• 2013

Background To construct a sophisticated three-dimensional framework, numerous modifications have been reported in the literature. However, most surgeons have paid little attention to the anatomical configuration of the concha and more to its deepness and hollowness, leading to unsatisfactory outcomes. Methods For a configuration of the concha that is definitely anatomical, the author further developed and employed the conchal bowl element, which has been used by several surgeons although the results have not been published elsewhere. The author constructed the conchal bowl element in one of three patterns according to the amount of available cartilages: one block, two-pieces, or a cymba bowl element only. A total of 20 patients underwent auricular reconstruction using a costal cartilage framework between 2009 and 2012. The 8 earliest reconstructions were performed without a conchal bowl element and the latter 12 with a conchal bowl element. The patients were followed up for more than 1 year. The aesthetic results were scored by evaluating characteristics involving the stability of the crus helicis, the conchal definition, and the smoothness of the helical curve. Results The ears reconstructed early without a conchal bowl element showed a shallow and one or two incompletely separated concha with an obliterated cymba conchal space. They also did not have a realistic or smooth curve of the helix because of an unstable crus helicis. However, ears reconstructed later with the concha bowl element showed a definite crus helicis, deep cymba conchal space, and smooth helical curve. Conclusions The construction of the conchal bowl element is simple, not time-consuming procedure. It is suggested that the conchal bowl element must be constructed and attached to the main framework for natural configuration of the reconstructed ear.

• Archives of Plastic Surgery
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• v.39 no.4
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• pp.323-328
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• 2012

Background Among the various methods for correcting nasal deformity, the composite graft is suitable for the inner and outer reconstruction of the nose in a single stage. In this article, we present our technique for reconstructing the ala and columella using the auricular chondrocutaneous composite graft. Methods From 2004 to 2011, 15 cases of alar and 2 cases of columellar reconstruction employing the chondrocutaneous composite graft were studied, all followed up for 3 to 24 months (average, 13.5 months). All of the patients were reviewed retrospectively for the demographics, graft size, selection of the donor site and outcomes including morbidity and complications. Results The reasons for the deformity were burn scar (n=7), traumatic scar (n=4), smallpox scar (n=4), basal cell carcinoma defect (n=1), and scar contracture (n=1) from implant induced infection. In 5 cases of nostril stricture and 6 cases of alar defect and notching, composite grafts from the helix were used ($8.9{\times}12.5$ mm). In 4 cases of retracted ala, grafts from the posterior surface of the concha were matched ($5{\times}15$ mm). For the reconstruction of the columella, we harvested the graft from the posterior scapha ($9{\times}13.5$ mm). Except one case with partial necrosis and delayed healing due to smoking, the grafts were successful in all of the cases and there was no deformity of the donor site. Conclusions An alar and columellar defect can be reconstructed successfully with a relatively large composite graft without donor site morbidity. The selection of the donor site should be individualized according to the 3-dimensional configuration of the defect.

• Proceedings of the Korea Crystallographic Association Conference
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• 2002.11a
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• pp.4-4
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• 2002

PDZ domains bind to short segments within target proteins in a sequence-specific fashion. GRIP/ABP family proteins contain six to seven PDZ domains and interact via its sixth PDZ domain (class Ⅱ) with the C-termini of various proteins, including liprin-α. In addition the PDZ456 domain mediates the formation of homo- and heteromultimers of GRIP proteins. To better understand the structural basis of peptide recognition by a class Ⅱ PDZ domain and DZ-mediated multimerization, we determined the crystal structures of the GRIPI PDZ6 domain, alone and in complex with a synthetic C-terminal octapeptide of human liprin-α, at resolutions of 1.5 Å and 1.8 Å, respectively. Remarkably, unlike other class Ⅱ PDZ domains, Ile736 at αB5 rather than conserved Leu732 at αB1 makes a direct hydrophobic contact with the side chain of the Tyr at the -2 position of the ligand. Moreover, the peptide-bound structure of PDZ6 shows a slight reorientation of helix αB, indicating that the second hydrophobic pocket undergoes a conformational adaptation to accommodate the bulkiness of the Tyr's side chain, and forms an antiparallel dimer through an interface located at a site distal to the peptide-binding groove. This configuration may enable formation of GRIP multimers and efficient clustering of GRIP-binding proteins.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.8
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• pp.1186-1194
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• 2019

Objective: The influence of broiler carcass scalding conditions on chicken breast meat quality parameters was investigated. Methods: Two hundred and seventy Cobb broiler chickens from 42 to 48 days old were slaughtered according to the standard industry practice and scalded in five temperature/time combinations-$T_1$, $54^{\circ}C/210s$; $T_2$, $55^{\circ}C/180s$; $T_3$, $56^{\circ}C/150s$; $T_4$, $57^{\circ}C/120s$; $T_5$, $58^{\circ}C/90s$. Results: Scalding temperature increase resulted in higher values of external and ventral lightness and in protein functionality reduction-determined by emulsification capacity and protein denaturation-in chicken breast fillets 24 h post-mortem. Protein secondary structures had conformational changes, with a decrease of the ${\alpha}$-helix and an increase of the ${\beta}$-sheet and ${\beta}$-turn proportions, mainly in $T_1$ and $T_5$ samples, determined by Fourier-transform infrared spectroscopy in an attenuated reflectance mode analysis. The chemical composition, pH, water holding capacity and Warner-Bratzler shear force did not differ among the treatments. In the fatty acid profile, the 18:1n-9 was lower in $T_5$, which suggested that the high scalding-temperature could have caused the lipid oxidation. The values of the polyunsaturated fatty acids (PUFA), such as 22:2, 20:4n-6, and 22:6n-3, were highest in the $T_5$, thus being related to the phospholipid cellular membrane collapse in this experimental condition and subsequent release of these PUFA. Conclusion: Intermediate scalding-parameters avoided the negative changes in the chicken meat quality.