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In reconstructive surgery, complications post-fibula free flap (FFF) reconstruction, notably peri-implant hyperplasia, are significant yet understudied. This study analyzed peri-implant hyperplastic tissue surrounding FFF, alongside peri-implantitis and foreign body granulation (FBG) tissues from patients treated at the Department of Oral and Maxillofacial Surgery, Seoul National University Dental Hospital. Using light microscopy, pseudoepitheliomatous hyperplasia, anucleate and pyknotic prickle cells, and excessive collagen deposition were observed in FFF hyperplastic tissue. Ultrastructural analyses revealed abnormal structures, including hemidesmosome dilation, bacterial invasion, and endoplasmic reticulum (ER) swelling. In immunohistochemical analysis, unfolded protein-response markers ATF6, PERK, XBP1, inflammatory marker NFκB, necroptosis marker MLKL, apoptosis marker GADD153, autophagy marker LC3, epithelial-mesenchymal transition, and angiogenesis markers were expressed variably in hyperplastic tissue surrounding FFF implants, peri-implantitis, and FBG tissues. NFκB expression was higher in peri-implantitis and FBG tissues compared to hyperplastic tissue surrounding FFF implants. PERK expression exceeded XBP1 significantly in FFF hyperplastic tissue, while expression levels of PERK, XBP1, and ATF6 were not significantly different in peri-implantitis and FBG tissues. These findings provide valuable insights into the interconnected roles of ER stress, necroptosis, apoptosis, and angiogenesis in the pathogenesis of oral pathologies, offering a foundation for innovative strategies in dental implant rehabilitation management and prevention., (© 2024. The Author(s).)

Objectives: The aim of this study was to perform a comparative analysis of the ultrastructural and chemical composition of sialoliths, tonsilloliths, and antroliths and to describe their growth pattern., Materials and Methods: We obtained 19 specimens from 18 patients and classified the specimens into three groups: sialolith (A), tonsillolith (B), and antrolith (C). The peripheral, middle, and core regions of the specimens were examined in detail by histology, micro-computed tomography (micro- CT), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy, and transmission electron microscopy (TEM)., Results: In the micro-CT, group A showed alternating radiodense and radiolucent layers, while group B had a homogeneous structure. Group C specimens revealed a compact homogeneous structure. Histopathologically, group A showed a laminated, teardrop-shaped, globular structure. Group B demonstrated degrees of immature calcification of organic and inorganic materials. In group C, the lesion was not encapsulated and showed a homogeneous lamellar bone structure. SEM revealed that group A showed distinct three layers: a peripheral multilayer zone, intermediate compact zone, and the central nidus area; groups B and C did not show these layers. The main elemental components of sialoliths were O, C, Ca, N, Cu, P, Zn, Si, Zr, F, Na, and Mg. In group B, a small amount of Fe was found in the peripheral region. Group C had a shorter component list: Ca, C, O, P, F, N, Si, Na, and Mg. TEM analysis of group A showed globular structures undergoing intra-vesicular calcification. In group B, bacteria were present in the middle layer. In the outer layer of the group C antrolith, an osteoblastic rimming was observed., Conclusion: Sialoliths had distinct three layers: a peripheral multilayer zone, an intermediate compact zone and the central nidus area, while the tonsillolith and antrolith specimens lacked distinct layers and a core.

According to the previous studies of sialolithiasis reported so far, this study is aimed to identify the biological components of sialolith, which show different ultrastructures and chemical compositions from other stones, cholelith and urolith. Twenty-two specimens obtained from 20 patients were examined histologically, and analyzed with micro-CT, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), and transmission electron microscopy (TEM). All sialoliths (n = 22) observed in this study showed a central nidus, which was filled with organoid matrix admixed with exosome vesicles, loose calcium apatite crystals, and many bacteria. The micro-CT and SEM observation clearly defined a single or multiple central nidus(es) encircled by highly calcified compact zone. The circular compact zone showed a band-like calcification, about 1-3 mm in thickness, and usually located between the central nidus and the peripheral multilayer zone. But some sialoliths (n = 5) showed severe erosion of compact zone by expanding multilayered zone depending on the level of calcification and inflammation in sialolith. By observing TEM images, many exosome vesicles and degraded cytoplasmic organelles were found in the central nidus, and some epithelial cells were also found in the calcified matrix of peripheral multilayer zone. Particularly, EDS analysis indicated the highest Ca/P ratio in the intermediate compact zone (1.77), and followed by the central nidus area (1.39) and the peripheral multilayer zone (0.87). Taken together, these data suggest that the central nidus containing many inflammatory exosomes and degraded cytoplasmic organelles has a potential to induce a band-like calcification of compact zone, and followed by the additional multilayer deposition of exfoliated salivary epithelial cells as well as salivary materials. Thereby, the calcium apatite-based sialolith is gradually growing in its volume size, and eventually obstructs the salivary flow and provides a site for the bacterial infection., (© 2023. The Author(s).)

Background: This study aimed to analyze the accuracy of the output of three-dimensional (3D) customized surgical guides and titanium implants in a rabbit model, and of mandibulectomy, reconstructive surgery, and surgical outcome; additionally, the correlation between surgical accuracy and surgical outcomes, including the differences in surgical outcome according to surgical accuracy, was analyzed., Results: The output of implants was accurately implemented within the error range (- 0.03-0.03 mm), and the surgical accuracy varied depending on the measured area (range - 0.4-1.1 mm). Regarding surgical outcomes, angle between the mandibular lower borders showed the most sensitive results and distance between the lingual cusps of the first molars represented the most accurate outcomes. A significant correlation was noted between surgical accuracy in the anteroposterior length of the upper borders pre- and postoperatively and the angle between the mandibular lower borders (regression coefficient = 0.491, p = 0.028). In the group wherein surgery was performed more accurately, the angle between the mandibular lower borders was reproduced more accurately (p = 0.021). A selective laser melting machine accurately printed the implants as designed. Considering the positive correlation among surgical accuracy in the mandibular upper borders, angle between the mandibular lower borders, and more accurately reproduced angle between the mandibular lower borders, the angle between the mandibular lower borders is considered a good indicator for evaluating the outcomes of reconstructive surgery., Conclusion: To reduce errors in surgical outcomes, it is necessary to devise a positioner for the surgical guide and design a 3D surgical guide to constantly maintain the direction of bone resection. A fixed area considering the concept of three-point fixation should be selected for stable positioning of the implant; in some cases, bilateral cortical bone fixation should be considered. The angle between the mandibular lower borders is a sensitive indicator for evaluating the outcomes of reconstructive surgery., (© 2023. The Author(s).)

Backgruound: A group health service is a system that delegates workplace health management to an entrusted institution. There have been various studies on group health services to date, but recent changes, such as an increase in foreign workers, are rapidly changing industry characteristics., Methods: Satisfaction was assessed using a 27-question survey distributed among 203 workplaces employing health professionals. The survey items consisted of general characteristics, comprehensive satisfaction, requirements for health professionals' work, and satisfaction with work environment management, ergonomic management, and healthcare management. Multiple regression and frequency analyses were performed., Results: The comprehensive satisfaction was 4.08 points on average, out of 5. The comprehensive satisfaction of health professionals in the industry was positively correlated with each factor. Hazardous materials and chemical management (material safety data sheets, MSDSs) were the most common requirements., Conclusion: A low level of satisfaction with work environment management indicates high demand for healthcare management. The working environment should be improved by identifying characteristics of the workplace, examining harmful substances, inspecting equipment, and enhancing worker methods. The shorter the work experience of health professionals, the more dependent they are on group health services. The variables affecting comprehensive satisfaction were the period of work, healthcare management satisfaction, and work environment management satisfaction. Most of the requirements of health professionals in the workplace were practical improvement case presentations, MSDSs, and legal document management.

Background: 4-Hexylresorcinol (4HR) is able to increase angiogenesis. However, its molecular mechanism in the human endothelial cells has not been clarified., Methods: As endothelial cells are important in angiogenesis, we treated the human umbilical vein endothelial cells (HUVECs) with 4HR and investigated protein expressional changes by immunoprecipitation high-performance liquid chromatography (IP-HPLC) using 96 antisera., Results: Here, we found that 4HR upregulated transforming growth factor-β (TGF-β)/SMAD/vascular endothelial growth factor (VEGF) signaling, RAF-B/ERK and p38 signaling, and M2 macrophage polarization pathways. 4HR also increased expression of caspases and subsequent cellular apoptosis. Mechanistically, 4HR increased TGF-β1 production and subsequent activation of SMADs/VEGFs, RAF-B/ERK and p38 signaling, and M2 macrophage polarization., Conclusion: Collectively, 4HR activates TGF-β/SMAD/VEGF signaling in endothelial cells and induced vascular regeneration and remodeling for wound healing., Competing Interests: Competing interestsThe authors declare that the authors have no competing interests as defined by Nature Research or other interests that might be perceived to influence the results and/or discussion reported in this paper., (© The Author(s) 2020.)

Allelic differences in A and B mating-type loci are a prerequisite for the progression of mating in the genus Pleurotus eryngii ; thus, the crossing is hampered by this biological barrier in inbreeding. Molecular markers linked to mating types of P. eryngii KNR2312 were investigated with randomly amplified polymorphic DNA to enhance crossing efficiency. An A4 -linked sequence was identified and used to find the adjacent genomic region with the entire motif of the A locus from a contig sequenced by PacBio. The sequence-characterized amplified region marker 7-2 299 distinguished A4 mating-type monokaryons from KNR2312 and other strains. A BLAST search of flanked sequences revealed that the A4 locus had a general feature consisting of the putative HD1 and HD2 genes. Both putative HD transcription factors contain a homeodomain sequence and a nuclear localization sequence; however, valid dimerization motifs were found only in the HD1 protein. The ACAAT motif, which was reported to have relevance to sex determination, was found in the intergenic region. The SCAR marker could be applicable in the classification of mating types in the P. eryngii breeding program, and the A4 locus could be the basis for a multi-allele detection marker.

Background: Alcoholic ketoacidosis (AKA) is known as a benign disease, but the related mortality reported in Korea is high. Acidosis and alcohol change the immunity profile, and these changes can be identified early using the delta neutrophil index (DNI). We aimed to evaluate the use of DNI and other standard laboratory parameters as predictors of prognosis in AKA patients., Methods: One hundred eighteen males with AKA were evaluated at the Wonju Severance Christian hospital between 2009 and 2014. We performed a retrospective analysis of demographic, clinical, and laboratory parameters data. Receiver operating characteristic curves (ROC) and multivariate Cox regression was used to identify renal survival and mortality., Results: Survival patients had lower initial DNI levels than non-survival patients (4.8±6.4 vs 11.4±12.5, p<0.001). In multivariate-adjusted Cox regression analysis, higher initial increased DNI (HR 1.044, 95% CI 1.003-1.086, p=0.035), and lower initial pH (HR 0.044, 95% CI 0.004-0.452, p=0.008) were risk factors for dialysis during hospitalization. Further, higher initial DNI level (HR 1.037; 95% CI 1.006-1.069; p=0.018), lower initial pH (HR 0.049; 95% CI 0.008-0.312; p=0.001) and lower initial glomerular filtration rate (GFR) (HR 0.981; 95% CI 0.964-0.999; p=0.033) were predictors of mortality. A DNI value of 4.5% was selected as the cut-off value for poor prognosis and Kaplan-Meier plots showed that AKA patients with an initial level DNI ≥4.5% had lower cumulative survival rates than AKA patients with an initial DNI <4.5%., Conclusion: Increased initial serum DNI levels may help to predict renal survival and prognosis in male AKA patients., Competing Interests: Conflict of interest: The material is not published previously, and will not be submitted for publication elsewhere.

Background: The purpose of this study was to evaluate the effects of botulinum toxin A (BTX) injection into the anterior belly of the digastric muscle on a growing rat., Methods: Ten Sprague Dawley rats were used in this study. When the rats were 13 days old, 0.5 units of BTX was injected into the anterior belly of the digastric muscle for the experimental group ( n  = 5). For the control, the same volume of normal saline was injected ( n  = 5). The rats were sacrificed at 60 days old, and the skulls were harvested for micro-computed tomography (μCT) analysis., Results: In anthropometric analysis, the zygomatic arch and mandibular bi-condylar width were significantly lower in the experimental group than those in the control group ( P  = 0.025 and 0.027, respectively). The maxillary point width was significantly higher in the experimental group than that in the control group ( P  = 0.020)., Conclusion: BTX injection into the anterior belly of the digastric muscle had effects on the maxillofacial bony width in growing rats., Competing Interests: This study was approved by the Institutional Animal Care and Use Committee of the Gangneung-Wonju National University, Gangneung, Korea (2016-24).This manuscript does not contain any individual person’s identifier (including individual details, images or videos).The authors declare that the authors have no competing interests as defined by Nature Research, or other interests that might be perceived to influence the results and/or discussion reported in this paper. KSG, KMK, and SH are editorial board members of “Maxillofacial Plastic and Reconstructive Surgery”.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

4-Hexylresorcinol (4HR) is a small organic compound that is used as an additive antiseptic and antioxidant, but its molecular properties have not been clearly elucidated. The present study explored the cellular effects of 4HR on RAW 264.7 cells by immunoprecipitation high-performance liquid chromatography (IP-HPLC) using 216 antisera. 4HR-treated cells showed significant decreases in the expressions of proliferation-related proteins, cMyc/MAX/MAD network, p53/Rb/E2F and Wnt/β-catenin signalings, epigenetic modifications, and protein translation. Furthermore, 4HR suppressed the expressions of growth factors and proteins associated with RAS signaling, NFkB signaling, inflammation, and osteogenesis, but elevated the expressions of proteins associated with p53-mediated and FAS-mediated apoptosis, T-cell immunity, angiogenesis, antioxidant, and oncogenic signaling. In a 4HR adherence assay, TNFα, PKC, osteopontin, and GADD45 were strongly adherent to 4HR-coated beads, whereas IL-6, c-caspase 3, CDK4, and c-caspase 9 were not. Many 4HR adherent proteins were expressed at lower levels in 4HR treated RAW 264.7 cells than in non-treated controls, whereas 4HR non-adherent proteins were expressed at higher levels. These observations suggest 4HR affects the expressions of proteins in an adhesion-dependent manner and that its effects on proteins are characteristic and global in RAW 264.7 cells.

Background: Coffee extract has been investigated by many authors, and many minor components of coffee are known, such as polyphenols, diterpenes (kahweol and cafestol), melanoidins, and trigonelline, to have anti-inflammatory, anti-oxidant, anti-angiogenic, anticancer, chemoprotective, and hepatoprotective effects. Therefore, it is necessary to know its pharmacological effect on hepatocytes which show the most active cellular regeneration in body., Methods: In order to determine whether coffee extract has a beneficial effect on the liver, 20 C57BL/6J mice were intraperitoneally injected once with dialyzed coffee extract (DCE)-2.5 (equivalent to 2.5 cups of coffee a day in man), DCE-5, or DCE-10, or normal saline (control), and then followed by histological observation and IP-HPLC (immunoprecipitation high performance liquid chromatography) over 24 h., Results: Mice treated with DCE-2.5 or DCE-5 showed markedly hypertrophic hepatocytes with eosinophilic cytoplasms, while those treated with DCE-10 showed slightly hypertrophic hepatocytes, which were well aligned in hepatic cords with increased sinusoidal spaces. DCE induced the upregulations of cellular proliferation, growth factor/RAS signaling, cellular protection, p53-mediated apoptosis, angiogenesis, and antioxidant and protection-related proteins, and the downregulations of NFkB signaling proteins, inflammatory proteins, and oncogenic proteins in mouse livers. These protein expression changes induced by DCE were usually limited to the range ± 10%, suggesting murine hepatocytes were safely reactive to DCE within the threshold of physiological homeostasis. DCE-2.5 and DCE-5 induced relatively mild dose-dependent changes in protein expressions for cellular regeneration and de novo angiogenesis as compared with non-treated controls, whereas DCE-10 induced fluctuations in protein expressions., Conclusion: These observations suggested that DCE-2.5 and DCE-5 were safer and more beneficial to murine hepatocytes than DCE-10. It was also found that murine hepatocytes treated with DCE showed mild p53-mediated apoptosis, followed by cellular proliferation and growth devoid of fibrosis signaling (as determined by IP-HPLC), and subsequently progressed to rapid cellular regeneration and wound healing in the absence of any inflammatory reaction based on histologic observations., Competing Interests: This study was approved by the Institutional Animal Care and Use Committee of the Gangneung-Wonju National University, Gangneung, South Korea (GWNU-2017-16).Not applicableThe authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

RAW 264.7 cells and HUVECs were compared to evaluate the effects of dialyzed coffee extract (DCE) and artificial coffee (AC). Immunoprecipitation high performance liquid chromatography (IP-HPLC) showed DCE-2.5- (equivalent to 2.5 cups of coffee a day) and DCE-5-induced protein expression that was beneficial to human health, i.e., they led to significant increases in proliferation-, immunity-, cellular protection-, antioxidant signaling-, and osteogenesis-related proteins but decreases in inflammation-, NFkB signaling-, cellular apoptosis-, and oncogenic signaling-related proteins in RAW 264.7 cells, and slight decreases in angiogenesis-related proteins in HUVECs. These protein expression changes were less frequently observed for DCE-10 treatment, while AC treatment induced very different changes in protein expression. We suggest that the favorable cellular effects of DCE were derived from minor coffee elements that were absent in AC, and that the reduced effects of DCE-10 compared with those of DCE-2.5 or DCE-5 might have been caused by greater adverse reactions to caffeine and chlorogenic acid in DCE-10 than DCE-2.5 or DCE-5. IP-HPLC results suggested that minor coffee elements in DCE might play beneficial roles in the global protein expression of proliferation-, immunity-, anti-inflammation-, cell protection-, antioxidant-, anti-apoptosis-, anti-oncogenesis-, and osteogenesis-related proteins in RAW 264.7 cells and enhance anti-angiogenic signaling in HUVECs.

Background: Calcium pyrophosphate dihydrate deposition disease (CPDD) is a rare disease in the temporomandibular joint (TMJ) space. It forms a calcified crystal mass and induces a limitation of joint movement., Case Presentation: The calcified mass in our case was occupied in the left TMJ area and extended to the infratemporal and middle cranial fossa. For a complete excision of this mass, we performed a vertical ramus osteotomy and resected the mass around the mandibular condyle. The calcified mass in the infratemporal fossa was carefully excised, and the segmented mandible was anatomically repositioned. Scanning electronic microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDS) microanalysis was performed to evaluate the calcified mass. The result of SEM/EDS showed that the crystal mass was completely composed of calcium pyrophosphate dihydrate. This result strongly suggested that the calcified mass was CPDD in the TMJ area., Conclusions: CPDD in the TMJ is a rare disease and is difficult to differentially diagnose from other neoplasms. A histological examination and quantitative microanalysis are required to confirm the diagnosis. In our patient, CPDD in the TMJ was successfully removed via the extracorporeal approach. SEM/EDS microanalysis was used for the differential diagnosis., Competing Interests: This case report was approved by the institutional review board (IRB) of Gangneung-Wonju National University Dental Hospital (2017-018).The patient consented to the publication of this case report.All authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Background: The objective of this study was to evaluate the influence of masticatory muscle injection of botulinum toxin type A (BTX-A) on the growth of the mandibular bone in vivo., Methods: Eleven Sprague-Dawley rats were used, and BTX-A ( n  = 6) or saline ( n  = 5) was injected at 13 days of age. All injections were given to the right masseter muscle, and the BTX-A dose was 0.5 units. All of the rats were euthanized at 60 days of age. The skulls of the rats were separated and fixed with 10% formalin for micro-computed tomography (micro-CT) analysis., Results: The anthropometric analysis found that the ramus heights and bigonial widths of the BTX-A-injected group were significantly smaller than those of the saline-injected group ( P  < 0.05), and the mandibular plane angle of the BTX-A-injected group was significantly greater than in the saline-injected group ( P  < 0.001). In the BTX-A-injected group, the ramus heights II and III and the mandibular plane angles I and II showed significant differences between the injected and non-injected sides ( P  < 0.05). The BTX-A-injected side of the mandible in the masseter group showed significantly lower mandibular bone growth compared with the non-injected side., Conclusion: BTX-A injection into the masseter muscle influences mandibular bone growth., Competing Interests: This study was approved by Gangneung-Wonju National University. The animal experiment was approved by the Institutional Animal Care and Use Committee of Gangneung-Wonju National University, Gangneung, Korea (IACUC GWNU-2016-24). All experiments were performed in accordance with the relevant guidelines and regulations.Not applicable.Authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

A patient had a right mandibular defect due to resection of an ameloblastoma. Previously, the defect had been reconstructed by an iliac bone graft, and subsequently, a titanium mesh with xenograft was used. However, it was not successfully reconstructed. For the recovery of mandible continuity and rehabilitation of jaw movement, we manufactured a customized 3-dimensional titanium implant by computer-aided design and manufacturing and electron beam melting technology. This implant was designed to have a porous body structure and lingual plate. The customized implant was accurately inserted in the bony defect. As a result, the patient showed a normal range of mouth opening and jaw movement. New bone migration was observed in the porous structure of the implant. Although there was a slight plate exposure and lack of alveolar bone formation, the customized 3D titanium implant successfully reconstructed the mandibular discontinuous defect and recovered jaw movement.

To understand the osteogenic effect of the middle layer of the silk cocoon, sericin was examined for its cellular effects associated with tumor necrosis factor-α (TNF-α) signaling in this study. The fragmented sericin proteins in the silk mat were evaluated for the TNF-α expression level in murine macrophages. The concentration of protein released from silk mats was higher in the outermost and the innermost layers than in the middle layers, and the protein released from the silk mat was identified as sericin. The level of TNF-α in murine macrophages was dependent on the applied concentration of sericin, and the expression of genes associated with osteogenesis in osteoblast-like cells was dependent on the applied concentration of TNF-α. In animal experiments, silk mats from the middle layers led to a higher regenerated bone volume than silk mats from the innermost layer or the outermost layer. If TNF-α protein was incorporated into the silk mats from the middle layers, bone regeneration was suppressed compared with unloaded silk mats from the middle layers. Accordingly, silk mats from the silk cocoon can be considered to be a fragmented sericin-secreting carrier, and the level of sericin secretion is associated with TNF-α induction and bone regeneration.

Background: Oral squamous cell carcinoma (OSCC) is one of the most dangerous cancers in the body, producing serious complications with individual behaviors. Many different pathogenetic factors are involved in the carcinogenesis of OSCC. Cancer cells derived from oral keratinocytes can produce different carcinogenic signaling pathways through differences in protein expression, but their protein expression profiles cannot be easily explored with ordinary detection methods., Methods: The present study compared the protein expression profiles between two different types of OSCCs, which were analyzed through immunoprecipitation high-performance liquid chromatography (IP-HPLC)., Results: Two types of squamous cell carcinoma (SCC) occurred in a mandibular (SCC-1) and maxillary gingiva (SCC-2), but their clinical features and progression were quite different from each other. SCC-1 showed a large gingival ulceration with severe halitosis and extensive bony destruction, while SCC-2 showed a relatively small papillary gingival swelling but rapidly grew to form a large submucosal mass, followed by early cervical lymph node metastasis. In the histological observation, SCC-1 was relatively well differentiated with a severe inflammatory reaction, while SCC-2 showed severely infiltrative growth of each cancer islets accompanied with a mild inflammatory reaction. IP-HPLC analysis revealed contrary protein expression profiles analyzed by 72 different oncogenic proteins. SCC-1 showed more cellular apoptosis and invasive growth than SCC-2 through increased expression of caspases, MMPs, p53 signaling, FAS signaling, TGF-β1 signaling, and angiogenesis factors, while SCC-2 showed more cellular growth and survival than SCC-1 through the increased expression of proliferating factors, RAS signaling, eIF5A signaling, WNT signaling, and survivin., Conclusions: The increased trends of cellular apoptosis and invasiveness in the protein expression profiles of SCC-1 were implicative of its extensive gingival ulceration and bony destruction, while the increased trends of cellular proliferation and survival in the protein profile of SCC-2 were implicative of its rapid growing tumor mass and early lymph node metastasis. These analyses of the essential oncogenic protein expression profiles in OSCC provide important information for genetic counseling or customized gene therapy in cancer treatment. Therefore, protein expression profile analysis through IP-HPLC is helpful not only for the molecular genetic diagnosis of cancer but also in identifying target molecules for customized gene therapy in near future.

Purpose: The objective of this study was to evaluate whether silk fibroin (SF) incorporated into 4-hexylresorcinol (4HR) could increase botulinum toxin-A (BTX-A) activity., Material and Methods: In total, 30 rats were used for this study. The animals were divided into 6 groups according to the injected materials (SA: saline only; SF; 4HR; B2: 2 units of BTX-A; B2 + SF + 4HR: combination of B2, SF, and 4HR; B5: 5 units of BTX-A). Serial sonography was used for the evaluation of muscle thickness after injection. Immunohistochemical staining was used for the evaluation of myosin type II (myo2) and Bcl-2 protein expression., Results: The relative thickness of the masseter muscle in B2 group was 66.14% ± 4.55% to the preinjection level; in B2 + SF + 4HR group was 54.59% ± 4.83%, and in B5 group was 56.19% ± 8.28%. Any BTX-injected group showed significantly lower value of the relative muscle thickness compared to SA, SF, or 4HR group (P < 0.001 for all). The difference of relative muscle thickness between B2 group and B2 + SF + 4HR group was statistically significant (P < 0.001). The intensity of myo2 immunostaining in B5, B2, and B2 + SF + 4HR group was significantly higher than those in the other groups (P < 0.05)., Conclusions: When 2 units of BTX was incorporated to SF and 4HR, combination formula showed similar activity to those of 5 units of BTX.

RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae . The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii . The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii .

A material for ridge preservation should have dimensional stability to resist bio-degradation. This study was designed to compare bio-degradation of ridge preservation materials. Collagen plug was used as a positive control. Untreated, ethanol-treated, and 4-hexylresorcinol (4HR)-treated silk plugs were used for the experimental group. Each material underwent a scanning electron microscopic exam and a Fourier transform infrared (FT-IR) spectroscopic exam. Bio-degradation was evaluated by analyzing cylindrical bony defects in rabbit tibias. There were no prominent differences in microstructure among the silk plug groups. FT-IR exam demonstrated that the ethanol- and 4HR-treated silk plug groups had enhanced β-sheet structure. All silk plug groups exhibited significantly higher residual graft than the collagen plug group 4 weeks postoperative ( p  < 0.05). In conclusion, silk fibroin-based ridge preservation material was less bio-degradable than a collagen plug until at least 4 weeks after grafting., Competing Interests: The authors have no conflicts of interest.This animal experiment was approved by the Institutional Animal Care and Use Committee of the Gangneung-Wonju National University (GWNU-2016-23).

Many congenital and acquired defects occur in the maxillofacial area. The buccal fat pad flap (BFP) is a simple and reliable flap for the treatment of many of these defects because of its rich blood supply and location, which is close to the location of various intraoral defects. In this article, we have reviewed BFP and the associated anatomical background, surgical techniques, and clinical applications. The surgical procedure is simple and has shown a high success rate in various clinical applications (approximately 90%), including the closure of oroantral fistula, correction of congenital defect, treatment of jaw bone necrosis, and reconstruction of tumor defects. The control of etiologic factors, size of defect, anatomical location of defect, and general condition of patient could influence the prognosis after grafting. In conclusion, BFP is a reliable flap that can be applied to various clinical situations.

Silk suture material is primarily composed of silk fibroin and regarded as a non-resorbable material. It is slowly degraded by proteolysis when it is implanted into the body. 4-Hexylresorcinol (4HR) is a well-known antiseptic. In this study, the biodegradability of 4HR-incorporated silk sutures were compared to that of untreated silk sutures and polyglactin 910 sutures, a commercially available resorbable suture. 4HR-incorporated silk sutures exhibited anti-microbial properties. Matrix metalloproteinase (MMP) can digest a wide spectrum of proteins. 4HR increased MMP-2, -3, and -9 expression in RAW264.7 cells. MMP-2, -3, and -9 were able to digest not only silk fibroin but also silk sutures. Consequently, 59.5% of the 4HR-incorporated silk suture material remained at 11 weeks after grafting, which was similar to that of polyglactin 910 degradation (56.4% remained). The residual amount of bare silk suture material at 11 weeks after grafting was 91.5%. The expression levels of MMP-2, -3 and -9 were high in the 4HR-incorporated silk suture-implanted site 12 weeks after implantation. In conclusion, 4HR-treated silk sutures exhibited anti-microbial properties and a similar level of bio-degradation to polyglactin 910 sutures and induced higher expression of MMP-2, -3, and -9 in macrophages.

Mucoepidermoid carcinoma (MEC) is the most common type of malignant neoplasm in the minor salivary gland. The hard palate is a frequently involved site of MEC. The treatment of low-grade MEC on the hard palate is wide local resection with a tumor-free margin. In the present case, the maxillary defect was reconstructed using a buccal fat pad (BFP) flap, followed by application of 4-hexylresorcinol (4HR) ointment for 2 weeks. The grafted BFP successfully covered the tumor resection defect without tension and demonstrated complete re-epithelialization without any complications., Competing Interests: No potential conflict of interest relevant to this article was reported.

Objective: The objective of this study was to evaluate (1) the effect of 4-hexylresorcinol (4HR) application on TNF-α expression in RAW264.7 cells and (2) the effect of 4HR ointment on burn wound healing in a rat burn wound model., Materials and Methods: RAW264.7 cells were cultured in the presence of different concentrations of 4HR, and the attached cells were sampled for western blot analysis at each time point. Animal studies were conducted on male Wistar rats (n=24). Three treatment groups were evaluated in this study: (1) treatment with ointment alone (negative control), (2) treatment with ointment containing 0.2wt% 4HR, and (3) treatment with ointment containing 2wt% 4HR. For the assessment of wound healing, digital photographs were taken at 1, 5, and 14 days. Animals were sacrificed at 1, 5, and 14 days for histological examination or ELISA analysis., Results: Western blot analysis showed that TNF-α protein levels were reduced following treatment with 4HR at concentrations between 1 and 10μg/mL. In animal studies, the average denuded area 14 days after burn was smaller in the 2wt% 4HR treatment group compared with the control group (P=0.022). Immunohistochemical analysis revealed lower TNF-α expression in the 2wt% 4HR treatment group compared with the control group 14 days after burn (P=0.027)., Conclusion: 4HR reduced TNF-α expression in RAW264.7 cells and in the burn wounds of rats. Burn wounds dressed with ointment containing 4HR also exhibited rapid epithelization and collagen regeneration., (Copyright © 2016 Elsevier Ltd and ISBI. All rights reserved.)

The purpose of this study was to compare the early or late expression levels of p65, Bcl-2, and type II myosin and the frequency of TUNEL-positive nuclei in the rat masseter muscle after injection of different concentrations of botulinum toxin-A (BTX-A). We injected either 5 U or 10 U of BTX-A into both masseter muscles of the rat. As a control group, the same volume of saline was injected. After 2 or 12 weeks, the animals were sacrificed. Subsequently, a biopsy and immunohistochemical staining of the samples were performed using a p65, Bcl-2, or type II myosin antibody. Additionally, a TUNEL assay and transmission electron microscopic analysis were performed. The expression of p65, Bcl-2, and type II myosin increased significantly with increasing concentrations of BTX-A at 2 weeks after BTX-A injection (P < 0.05). The number of TUNEL-positive nuclei was also significantly increased in the BTX-A-treated groups in comparison to the saline-treated control at 2 weeks after BTX-A injection (P < 0.05). Elevated expression of Bcl-2 was also observed in 10-unit BTX-A-treated group at 12 weeks after injection (P < 0.05). At 12 weeks after injection, the number of enlarged mitochondria was increased, and many mitochondria displayed aberrations in cristae morphology after BTX-A injection. In conclusion, BTX-A injection into the masseter muscle increased the expression level of p65, Bcl-2, and type II myosin at an early stage. The morphological changes of mitochondria were more evident at 12 weeks after injection.

Background: Most of cleft lip and palate patients have the esthetic and functional problems of midfacial deficiencies due to innate developmental tendency and scar tissues from repeated operations. In these cases, maxillary protraction is required for the harmonious facial esthetics and functional occlusion., Case Presentation: A 7-year old boy had been diagnosed as severe maxillary constriction due to unilateral complete cleft lip and palate. The author tried to correct the secondary deformity by early distraction osteogenesis with the aim of avoiding marked psychological impact from peers of elementary school. From 1999 to 2006, repeated treatments, which consisted of Le Fort I osteotomy and face mask distraction, and complementary maxillary protraction using miniplates were performed including orthodontics. But, final facial profile was not satisfactory, which needs compromising surgery., Conclusions: The result of this study suggests that if early distraction treatment is performed before facial skeletal growth is completed, an orthognathic surgery or additional distraction may be needed later. Maxillofacial plastic and reconstructive surgeons should notify this point when they plan early distraction treatment for cleft maxillary deformity.

Background: Imperata cylindrica is being considered as a biomass candidate for bioethanol. This work aimed to evaluate a mild alkali pretreatment effect on the Imperata recalcitrant structure. Therefore, varied concentrations of NaOH (0, 7.5, 15, 20, and 25 g L(-1) ) were applied as treatments to Imperata at 105 °C for 10 min., Results: Scanning electron microscopy, atomic force microscopy, and Fourier transform infrared spectroscopy studies revealed that 20 to 25 g L(-1) NaOH-treated Imperata exposed amorphous cellulose on surface granules composed of lignin, waxes, and partly hemicelluloses were abolished due to the comprehensive disruption of the linkages between lignin and carbohydrates. The cellulose crystalline index was increased with 7.5 to 20 g L(-1) NaOH treatments and reduced with a 25 g L(-1) NaOH treatment. In fact, the cellulose content in solids increased with the increasing NaOH concentration and was estimated to be 720 and 740 g kg(-1) for the 20 and 25 g L(-1) NaOH treatments, respectively. The yield of the reducing sugar was obtained 805 and 813 mg g(-1) from 20 and 25 g L(-1) NaOH-treated Imperata, respectively., Conclusion: Considering the cost of pretreatment, the 20 g L(-1) NaOH treatment is judged to be effective for disrupting Imperata recalcitrance in this pretreatment regime., (© 2015 Society of Chemical Industry.)

Background: One of the most important members of the genus Brassica, cabbage, requires a relatively high level of calcium for normal growth (Plant Cell Environ 7: 397-405, 1984; Plant Physiol 60: 854-856, 1977). Localized Ca(2+) deficiency in cabbage leaves causes tip-burn, bringing about serious economic losses (Euphytica 9:203-208, 1960; Ann Bot 43:363-372, 1979; Sci Hortic 14:131-138, 1981). Although it has been known that the occurrence of tip-burn is related to Ca(2+) deficiency, there is limited information on the underlying mechanisms of tip-burn or the relationship between Ca(2+) and tip-burn incidence. To obtain more information on the genetic control of tip-burn symptoms, we focused on the identification of genes differentially expressed in response to increasing intracellular Ca(2+) and K(+) concentrations in B. oleracea lines derived from tip-burn susceptible, tip-burn resistant cabbages (B. oleracea var. capitata), and kale (B. oleracea var. acephala)., Results: We compared the levels of major macronutrient cations, including Ca(2+) and K(+), in three leaf segments, the leaf apex (LA), middle of leaf (LM), and leaf base (LB), of tip-burn susceptible, tip-burn resistant cabbages, and kale. Ca(2+) and K(+) concentrations were highest in kale, followed by tip-burn resistant and then tip-burn susceptible cabbages. These cations generally accumulated to a greater extent in the LB than in the LA. Transcriptome analysis identified 58,096 loci as putative non-redundant genes in the three leaf segments of the three B. oleracea lines and showed significant changes in expression of 27,876 loci based on Ca(2+) and K(+) levels. Among these, 1844 loci were identified as tip-burn related phenotype-specific genes. Tip-burn resistant cabbage and kale-specific genes were largely related to stress and transport activity based on GO annotation. Tip-burn resistant cabbage and kale plants showed phenotypes clearly indicative of heat-shock, freezing, and drought stress tolerance compared to tip-burn susceptible cabbages, demonstrating a correlation between intracellular Ca(2+) and K(+) concentrations and tolerance of abiotic stress with differential gene expression. We selected 165 genes that were up- or down-regulated in response to increasing Ca(2+) and K(+) concentrations in the three leaf segments of the three plant lines. Gene ontology enrichment analysis indicated that these genes participated in regulatory metabolic processes or stress responses., Conclusions: Our results indicate that the genes involved in regulatory metabolic processes or stress responses were differentially expressed in response to increasing Ca(2+) and K(+) concentrations in the B. oleracea leaf. Our transcriptome data and the genes identified may serve as a starting point for understanding the mechanisms underlying essential macronutrient deficiencies in plants, as well as the features of tip-burn in cabbage and other Brassica species.

Background: Silk cocoon is composed of multiple layers. The natural silk cocoon containing all layers was cut as a rectangular shape as defined as total group. The inner and outermost layers were removed from the total group and the remained mat was defined as the middle group. The objectives of this study was to compare the total group with the middle group as a barrier membrane for the guided bone regeneration., Methods: The effects of these materials on the cellular proliferation and alkaline phosphatase (ALP) expression of MG63 cells were explored. For comparing bone regeneration ability, bilateral bone defects were created in calvarial areas in ten adult New Zealand white rabbits. The defects were covered with silk membranes of the middle group, with silk membrane of the total group used as the control on the contralateral side. The defects were allowed to heal for 4 and 8 weeks. Micro-computerized tomography (μCT) and histological examination were performed., Results: The middle group exhibited a higher MTT value 48 and 72 h after treatment compared to the total group. ALP expression was also higher in the middle group. The results of μCT and histologic examination showed that new bone formation was significantly higher in the middle group compared to the total group 8 weeks postoperatively ( P  < 0.05)., Conclusions: In conclusion, the middle layer of the silk cocoon supports guided bone regeneration better than unprocessed silk cocoon.

Background: The purpose of this study was to evaluate the change of food intake after different dosages of botulinum toxin A (BTX) injection in the animal model. Additionally, the dimensional and histological change at 14 days after BTX injection was also evaluated., Methods: The comparative study was performed using the BTX injection model in rats ( n  = 5 for each group). Group 1 was the saline-injected group. Group 2 was the 5-unit BTX-injection group to each masseter muscle. Group 3 was the 10-unit BTX-injection group to each masseter muscle. Food intake rates and body weight were checked daily before and after BTX injection until 10 days. All animals were sacrificed at 14 days after BTX injection, and the specimens underwent hematoxylin and eosin stain and immunohistochemical staining for myosin type II (MYH2)., Results: The recovery of food intake in groups 2 and 3 decreased significantly compared with group 1 from day 2 to day 7 and day 9 after injection ( p  < 0.05). The BTX-treated masseter muscles were significantly smaller than those in group 1 ( p  = 0.015). The immunohistochemical findings demonstrated that the expression of MYH2 was significantly higher in group 3 compared to groups 1 and 2 ( p  < 0.001)., Conclusions: BTX injection to the masseter muscle in rats demonstrated short food-intake-rate reduction with recovery until 10 days after injection. The thickness of the masseter muscle and MYH2 expression were significantly changed according to the injected dose of BTX.

Gorham disease is an idiopathic massive osteolysis, and maxillofacial involvement is rare. This report describes a case of a 12-year-old boy with severe progressive osteolysis in the mandible, hyoid bone, mastoid process, and cervical spine. Radiation therapy and interferon-α therapy were followed by bisphosphonate therapy. The patient died of respiratory failure. To describe the clinicopathologic features of Gorham disease of the jaws with an emphasis on the fatal types, 64 cases in the literature were reviewed (female-to-male ratio, 1:1.78; average age, 33.02 ± 19.38 yr). Most lesions were located only in the mandible or in other locations in combination with the mandible, except for 3 cases. During follow-up, there were 7 cases of disease-specific death, resulting in a mortality rate of 10.94%. The main causes of death were chylothorax, rib fractures secondary to osteolysis, or spinal fractures. Although most patients received surgical treatment (43.75%), the type of treatment was not related to prognosis., (Copyright © 2015 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

After cleft lip repair, many patients suffer from nasolabial fistulas, asymmetrical nasal floor, or an indistinct nostril sill, as well as intraoral wound dehiscence and subsequent scar contracture of surgical wounds leading to vestibular stenosis. For successful primary nasolabial repair of complete cleft deformity of the primary palate, cleft surgeons need special care in reconstructing the sound nasal floor. Especially when the cleft gap is wide or when any type of nasoalveolar molding therapy was not performed, three-dimensional reconstruction of the nasal floor is critical for a balanced nasal shape. In this study, the author describes an effective method for reconstructing a double-layered nasal floor using two mucosal flaps from both sides of the fissured upper lip. This is a report of six patients with unilateral or bilateral complete cleft of the primary palate with a detailed description of the surgical technique and a literature review.

Hepatoblastoma usually occurs in children under the age of 2 years, with very few cases reported in adults. We experienced a case of adult hepatoblastoma in a 36-year-old female with chronic hepatitis B. She had experienced sudden onset abdominal pain. Her serum alpha-fetoprotein level was markedly elevated, and abdominal CT showed a 9-cm mass with internal hemorrhage in the right hepatic lobe with hemoperitoneum, so an emergency hepatic central bisectionectomy was performed. The initial histologic examination revealed that the mass mimicked combined hepatocellular carcinoma and cholangiocarcinoma with spindle-cell metaplasia of the cholangiocarcinoma element. Follow-up abdominal CT performed 3 months later showed a 5.5-cm metastatic mass in the left subphrenic area. Laparoscopic splenectomy with mass excision was performed, and hepatoblastoma was confirmed histologically. A histologic re-examination of previously obtained surgical specimens also confirmed the presence of hepatoblastoma. Metastatic hepatoblastoma was found at multiple sites of the abdomen during follow-up, and so chemotherapy with cisplatin, 5-fluorouracil (5-FU), and vincristine was applied, followed by carboplatin and doxorubicin. Despite surgery and postoperative chemotherapy, she died 12 months after symptom onset.

One complication related to local anesthesia in the dental clinic is a broken needle. Although rare, a broken needle may be difficult to retrieve. Dental radiographs and 3D CT have been used in the past to confirm the location of a broken needle. We present the case of a broken needle, which was successfully removed using a careful, microscopic approach.

Purpose: The purpose of this study was to evaluate the coverage of the pedicled buccal fat pad flap (PBFP) and the long-term results of this treatment in patients with medication-related osteonecrosis of the jaw (MRONJ)., Patients and Methods: Ten patients (2 men and 8 women; average age, 72.9 yr old) diagnosed with MRONJ were selected. Patients were treated with a PBFP. Data from patients regarding MRONJ stage, defect size, bone exposure after surgery, operation time, admission period, duration of antibiotic therapy, recurrence of disease, and postoperative complications were analyzed retrospectively., Results: Six patients were diagnosed with MRONJ stage 2, and 4 patients were diagnosed with MRONJ stage 3. The maximum defect in the study was 62 × 18 mm. Among the 10 patients, there was only 1 bony exposure, which occurred on postoperative day 2 after receiving the PBFP. This exposure might have been due to an incomplete resection of the affected bone. There were no severe donor site morbidities, and all patients showed satisfactory healing without incident., Conclusions: According to this evaluation, the PBFP effectively covered a relatively large surgical defect. Complications were minimal, and there was no recurrence of bony exposure during follow-up. In conclusion, using the PBFP was a reliable treatment option for the management of denuded bone in patients with MRONJ., (Copyright © 2015 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

To characterize of the pathogenicity gene from the soft rot pathogen Pantoea sp. PPE7 in Pleurotus eryngii, we constructed over 10,000 kanamycin-resistant transposon mutants of Pantoea sp. strain PPE7 by transposon mutagenesis. One mutant, Pantoea sp. NPPE9535, did not cause a soft rot disease on Pleurotus eryngii was confirmed by the pathogenicity test. The transposon was inserted into the wzc gene and the disruption of the wzc gene resulted in the reduction of polysaccharide production and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. Analysis of the hydropathic profile of this protein indicated that it is composed of two main domains: an N-terminal domain including two transmembrane α-helices and a C-terminal cytoplasmic domain consisting of a tyrosine-rich region. Comparative analysis indicated that the amino acid sequence of Wzc is similar to that of a number of proteins involved in the synthesis or export of polysaccharides in other bacterial species. Purified GST-Wzc was found to affect the phosphorylation of tyrosine residue in vivo. These results showed that the wzc gene might play an important role in the virulence of Pantoea sp. strain PPE7 in P. eryngii., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Purpose: This study evaluated woven silk textile for burn wound dressing materials in an animal model., Methods: Ten rats were used in this experiment. Full-thickness 2×2 cm burn wounds were created on the back of the rats under anesthesia. In the experimental group, the wounds were treated with three different dressing materials from woven silk textile. In the control group, natural healing without any dressing material was set as control. The wound surface area was measured at five days, seven days, and 14 days. Wound healing was evaluated by histologic analysis., Results: There were no statistically significant differences among groups at five days post injury. The mean defect size at seven days was largest in Group 3 (462.87 mm(2)), and smallest in Group 1 (410.89 mm(2)), not a significant difference (P =0.341). The mean defect size at 14 days was smallest at the Group 3 (308.28 mm(2)) and largest in the control group (388.18 mm(2)), not a significant difference (P =0.190). The denuded area was smaller in Group 1 (84.57 mm(2)) and Group 2 (82.50 mm(2)) compared with the control group (195.93 mm(2)), not statistically significant differences (P =0.066, 0.062). The difference between Group 3 and control was also not statistically significant (P =0.136). In histologic analysis, the experimental groups re-epithelialized more than control groups. No evidence was found of severe inflammation., Conclusion: The healing of burn wounds was faster with silk weave textile more than the control group. There was no atypical inflammation with silk dressing materials. In conclusion, silk dressing materials could be used to treat burn wounds.

The objective of this study was to compare the effectiveness of silk membranes (SMs) of different thicknesses for guided bone regeneration. Two kinds of SMs were prepared (SM1: 0.01 mm thickness, SM2: 0.5 mm thickness). Before use in animal experiments, scanning electron microscope images were taken to examine the gross morphology of each membrane. Ten New Zealand white rabbits were used for this study. Bilateral round-shaped defects were created in the parietal bone (diameter: 8.0 mm) and each defect was covered with SM1 or SM2. Animals were killed at 4 weeks and 8 weeks. Bone regeneration was analyzed in each specimen by micro-computed tomography (μ-CT) and histological analysis. In the μ-CT analysis, the average amount of newly formed bone in the SM2 group was greater than that in the SM1 group. There was a significant difference at 4 weeks after surgery (P = 0.004). In the histological analysis, the amount of formed lamellar bone was much greater in the SM2 group than in the SM1 group at 8 weeks after surgery (P = 0.021). In conclusion, the thick SM was much more effective for bone regeneration of bone defects than the thin SM.

The radial forearm free flap (RFFF) is a thin and pliable tissue with many advantages for tongue reconstruction. However, tongues reconstructed with RFFF occasionally need revision surgery because inadequate defect measurement at primary surgery can lead to bulkiness and limited movement of reconstructed tongue. In this case, the patient underwent partial glossectomy and RFFF reconstruction for treatment of tongue cancer five years prior. We could not make a lower denture for the patient, because the alveolo-lingual sulcus of tongue was almost lost. So we performed vestibuloplasty with a modified Kazanjian method on the lingual vestibule of the mandibular right posterior area, and defatting surgery to debulk the flap. After surgery, we observed that the color and texture of the revised tongue changed to become similar with adjacent tissue. The patient obtained a more functional and esthetic outcome. Accordingly, we present a case report with a review of relevant literature.

The molecular karyotype of Hypsizygus marmoreus was explored by contour-clamped homogeneous electric field gel electrophoresis. Eleven chromosomal bands were separated from the dikaryotic mycelia of H. marmoreus (strain Hm 3-10), and the chromosomes ranged in size from 1.9 to 5.8 Mb. The total genome size of the strain was estimated to be 36.3 Mb. The chromosome numbers were also confirmed by telomere fingerprinting, and 22 telomeric bands were identified. This result suggests that 11 chromosomes exist in Hm 3-10. The marker sequences for each chromosome were determined and were applied to identify each chromosome. Karyotyping and Southern blot analysis revealed that the size of chromosomes in the basidiospores were greatly different from those of parental dikaryon Hm 3-10 cells., (© 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)

Chlorpyrifos is an organophosphate pesticide that has adverse effect on animals and plants. We isolated endophytic bacterial strain, Pseudomonas sp. BF1-3, from balloon flower root which can hydrolyze chlorpyrifos. A gene (ophB) encoding a protein involved in chlorpyrifos degradation from this strain was cloned into Escherichia coli DH5α for confirming enzyme activity. After sequencing, total 1024bp nucleotide sequences were found in the open reading frame of ophB. The chlorpyrifos degradation patterns by E. coli DH5α (ophB) were observed. During incubation in minimal salt (M9) medium supplemented with chlorpyrifos (100mgL(-1)), the E. coli DH5α harboring ophB degraded about 97% initial chlorpyrifos (100mgL(-1)) and accumulated 86mgL(-1) 3,5,6-trichloro-2-pyridinol (TCP) within 9 days. In addition, optical density (OD) of E. coli DH5α (ophB) culture at 600nm was increased from 0.172 to 1.118 within 2 days of inoculation in the chlorpyrifos supplemented M9 medium. The estimated molecular weight of purified OphB protein was determined to be 31.4kDa by SDS-PAGE. The OphB enzyme was most active at pH 8 and an optimal temperature around 35°C. These results indicate that endophytic bacteria are supposed to be useful for biological control of environments contaminated with pesticides., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Pleurotus eryngii has recently become a major cultivated mushroom; it uses tetrapolar heterothallism as a part of its reproductive process. Sexual development progresses only when the A and B mating types are compatible. Such mating incompatibility occasionally limits the efficiency of breeding programs in which crossing within loci-shared strains or backcrossing strategies are employed. Therefore, understanding the mating system in edible mushroom fungi will help provide a short cut in the development of new strains. We isolated and identified pheromone and receptor genes in the B3 locus of P. eryngii and performed a functional analysis of the genes in the mating process by transformation. A genomic DNA library was constructed to map the entire mating-type locus. The B3 locus was found to contain four pheromone precursor genes and four receptor genes. Remarkably, receptor PESTE3.3.1 has just 34 amino acid residues in its C-terminal cytoplasmic region; therefore, it seems likely to be a receptor-like gene. Real-time quantitative RT-PCR (real-time qRT-PCR) revealed that most pheromone and receptor genes showed significantly higher expression in monokaryotic cells than dikaryotic cells. The pheromone genes PEphb3.1 and PEphb3.3 and the receptor gene PESTE3.3.1 were transformed into P5 (A3B4). The transformants were mated with a tester strain (A4B4), and the progeny showed clamp connections and a normal fruiting body, which indicates the proposed role of these genes in mating and fruiting processes. This result also confirms that PESTE3.3.1 is a receptor gene. In this study, we identified pheromone and receptor genes in the B3 locus of P. eryngii and found that some of those genes appear to play a role in the mating and fruiting processes. These results might help elucidate the mechanism of fruiting differentiation and improve breeding efficiency.

Background: The Brassicaceae family is an exemplary model for studying plant polyploidy. The Brassicaceae knowledge-base includes the well-annotated Arabidopsis thaliana reference sequence; well-established evidence for three rounds of whole genome duplication (WGD); and the conservation of genomic structure, with 24 conserved genomic blocks (GBs). The recently released Brassica rapa draft genome provides an ideal opportunity to update our knowledge of the conserved genomic structures in Brassica, and to study evolutionary innovations of the mesohexaploid plant, B. rapa., Results: Three chronological B. rapa genomes (recent, young, and old) were reconstructed with sequence divergences, revealing a trace of recursive WGD events. A total of 636 fast evolving genes were unevenly distributed throughout the recent and young genomes. The representative Gene Ontology (GO) terms for these genes were 'stress response' and 'development' both through a change in protein modification or signaling, rather than by enhancing signal recognition. In retention patterns analysis, 98% of B. rapa genes were retained as collinear gene pairs; 77% of those were singly-retained in recent or young genomes resulting from death of the ancestral copies, while others were multi-retained as long retention genes. GO enrichments indicated that single retention genes mainly function in the interpretation of genetic information, whereas, multi-retention genes were biased toward signal response, especially regarding development and defense. In the recent genome, 13,302, 5,790, and 20 gene pairs were multi-retained following Brassica whole genome triplication (WGT) events with 2, 3, and 4 homoeologous copies, respectively. Enriched GO-slim terms from B. rapa homomoelogues imply that a major effect of the B. rapa WGT may have been to acquire environmental adaptability or to change the course of development. These homoeologues seem to more frequently undergo subfunctionalization with spatial expression patterns compared with other possible events including nonfunctionalization and neofunctionalization., Conclusion: We refined Brassicaceae GB information using the latest genomic resources, and distinguished three chronologically ordered B. rapa genomes. B. rapa genes were categorized into fast evolving, single- and multi-retention genes, and long retention genes by their substitution rates and retention patterns. Representative functions of the categorized genes were elucidated, providing better understanding of B. rapa evolution and the Brassica genus.

Purpose: This study evaluated powdered burn wound dressing materials from wild silkworm fibroin in an animal model., Methods: Fifteen rats were used in this experiment. Full-thickness 2×2 cm burn wounds were created on the back of rats under anesthesia. In the two experimental groups, the wounds were treated with two different dressing materials made from silkworm fibroin. In the Control Group, natural healing without any dressing material was set as control. The wound surface area was measured at five days, seven days and 14 days. Wound healing was evaluated by histologic analysis., Results: By gross observation, there were no infections or severe inflammations through 14 days post-injury. The differences among groups were statistically significant at seven days and 14 days, postoperatively (P <0.037 and 0.001, respectively). By post hoc test, the defect size was significantly smaller in experimental Group 1 compared with the Control Group and experimental Group 2 at seven days postoperatively (P =0.022 and 0.029, respectively). The difference between Group 1 and Group 2 was statistically significant at 14 days postoperatively (P <0.001). Group 1 and control also differed significantly (P =0.002). Group 1 showed a smaller residual scar than the Control Group and Group 2 at 14 days post-injury. Histologic analysis showed more re-epithelization in Groups 1 and 2 than in the Control Groups., Conclusion: Burn wound healing was accelerated with silk fibroin spun by wild silkworm Antheraea pernyi. There was no atypical inflammation with silk dressing materials. In conclusion, silk dressing materials can be used for treatment of burn wound.

Gotgam chamoe (GgC), a native oriental melon in Korea, is known to possess the aroma of a dried persimmon, an agronomic relevance for melon breeding program. The volatile compounds and the transcript levels of aromatic compound genes in cultivar (Ohbokggul chamoe [OC]) and GgC were profiled. A total of 62 volatile compounds were identified and quantified. Twenty-eight volatile compounds were specific to either the OC or the GgC. The amounts of volatile alcohol, saturated hydrocarbon, and unsaturated hydrocarbon compounds were 2.2, 2.7, and 1.1 times higher in OC, respectively. The amounts of ketone volatiles were 1.2 times higher in GgC, whereas the total amounts of esters were similar. In the shikimate pathway, transcriptional patterns with the fruit parts were different between the two chamoes for CmDAHPS, CmDHD/SDH, and CmEPSPS. The expression levels of all six genes investigated, especially CmCS, were highest in the peel of both chamoes compared to the other parts. The transcript levels of the aromatic amino acid biosynthesis genes demonstrate that phenylalanine and tyrosine are present more in edible parts of the chamoe, while tryptophan may be accumulated low in the chamoe. In addition, phenylalanine and tryptophan are synthesized more in GgC than the OC.

In 2009-2010, unusual symptoms were observed on Pleurotus eryngii grown in mushroom farms in Gyeongnam Province, Republic of Korea. One of the main symptoms was a cobweb-like growth of fungal mycelia over the surface of the mushroom. The colonies on the surface rapidly overwhelmed the mushrooms and developed several spores within 3-4 days. The colonized surface turned pale brown or yellow. The fruit body eventually turned dark brown and became rancid. Koch's postulates were completed by spraying and spotting using isolated strains. The phylogenetic tree obtained from the internal transcribed spacer sequence analysis showed that the isolated fungal pathogen corresponded to Cladobotryum mycophilum (99.5%). In the fungicide sensitivity tests, the ED50 values for the isolate with respect to benomyl and carbendazim were from 0.29 to 0.31 ppm. Benzimidazole fungicides were most effective against C. mycophilum, a causal agent of cobweb disease in P. eryngii.

Purpose: The objective of this study was to compare peri-implant bone formation among uncoated (UC), hydroxyapatite (HA), collagen plus HA (CH), and collagen, HA, plus bone morphogenetic protein-2 (BMP-2) implant groups., Materials and Methods: Implants in the UC group had acid-etched surfaces. The surface coating was applied using the aerosol deposition method. The coated surfaces were examined by scanning electron microscopy, x-ray diffraction (XRD), and Fourier-transformed infrared absorption analysis. Subsequently, 6 implants from each group (total, 24 implants) were installed in the tibias of rabbits. The animals were sacrificed at 6 weeks after implant installation. Peri-implant bone formation and bone-to-implant contact (BIC) were measured in histologic sections. Significant differences among groups were evaluated using analysis of variance., Results: Based on the measured XRD patterns, there was a characteristic HA phase (International Centre for Diffraction Data [ICDD], 086-0740) coated on the titanium (ICDD, 089-3725). Subsequent coating processes for collagen and BMP-2 did not display additional diffraction peaks, but maintained the diffraction patterns of the HA-coated titanium. The presence of collagen was verified by infrared absorption analysis. When comparing these modifications with UC surfaces, only the CH coating displayed significantly greater peri-implant bone formation and BIC (P = .003 and P < .001, respectively). Adding BMP-2 to the implant surface did not produce any advantage compared with the CH coating., Conclusions: In this study, the CH group displayed significantly greater new bone formation and BIC than the other groups. There was no significant difference among the other groups., (Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.)

The effect of dilute sodium hydroxide (NaOH) on reed straw structural change at 105 degreeC temperature was evaluated in this study. Various concentrations of NaOH (1% to 2.5%) were used for pretreatment of reed straw at 105 degreeC for 10min. Scanning electron microscopy, atomic force microscopy and Fourier transform infrared spectroscopy studies showed that 2% and 2.5% NaOH pretreated sample exposed more cellulose fibers compared with other treatments. The cellulose crystalline index was increased by the 1% to 2.0% NaOH treatments and slightly lowered by the 2.5% NaOH treatment due to destructing cellulose fibres. Two per cent NaOH pretreatment caused 69.9% lignin removal, whereas 2.5% NaOH pretreatment removed 72.4% lignin. Besides, reed straw, when pretreated at 2% and 2.5% NaOH, resulted 56.4% and 60.5% hemicellulose removal, respectively. However, the difference in removal of lignin and hemicellulose between 2% and 2.5% NaOH treated reed straw was very marginal. In addition, very negligible increase of cellulose level was estimated, amounting 78.8% and 76.6% in 2.5% and 2% NaOH-treated sample, respectively. Moreover, after 72 h, reducing sugar yield was 81.2% and 83.3% using enzyme loading of 15 FPU (g dry biomass)-' and 30 IU (g dry biomass)- and xylanase 4 FXU (g dry biomass)-1 from 2% and 2.5% NaOH pretreated reed straw, respectively. Reducing sugar yield was increased very marginally when NaOH concentration increased from 2% to 2.5% for reed straw pretreatment. Therefore, 2% NaOH is supposed to be effective for reed straw pretreatment at this mentioned condition.

Background: The cyclosporin A (CsA) nephrotoxicity limits its usefulness as an immunosuppression. We studied the administration of both nitroprusside and phosphodiesterase-5 inhibitor (udenafil) in order to determine whether these agents could ameliorate the renal injury in CsA nephrotoxicity., Methods: 30 8-week-old SD rats were divided into 5 groups: the control (1), SQ with 15 mg/kg CsA (Group 2), CsA along with 5 mg/kg IP nitroprusside (Group 3), CsA with 10 mg/kg PO udenafil (Group 4), and CsA with udenafil and nitroprusside (Group 5)., Results: Group showed an increase in creatinine compared o the control group. Group 5 showed a decrease in creatinine compared to Group 2. In TUNEL, Group 2 increased apoptosis in proximal tubules compared to control. Group 5 showed a decrease in apoptosis compared to Groups 2, 3, and 4. In IHC, the eNOS in Group 2 was stronger than in the controls. Groups 3, 4, and 5 showed decreased staining intensity compared to Group 2. In IHC, the VEGF in Groups 2, 3, and 4 increased compared to the controls. The eNOS protein expression was increased in both Groups 3 and 5 compared to the controls. The VEGF protein expression was increased in Groups 3 and 5 compared to Group 2. The eNOS mRNA was decreased in Group 2 compared to the control group and tended to increase in Groups 3, 4, and 5 compared to Group 2. The VEGF mRNA was increased in Group 2 and tended to increase more in Groups 3 and 5., Conclusion: The udenafil and nitroprusside ameliorated renal injury in rat model of CsA nephrotoxicity. The mechanism appears to be associated with decreasing tubular apoptosis by decreasing eNOS and increasing VEGF.