Anti-fibrotic and anti-stricture effects of biodegradable biliary stents braided with dexamethasone-impregnated sheath/core structured monofilaments.

Endoscopic biliary stent insertion has been widely used for the treatment of benign biliary stricture (BBS). Thus, the development of stent materials in the perspectives of structure, mechanical properties, and biocompatibility has been also studied. However, conventional metal and plastic stents have several disadvantages, such as repeated procedures to remove or exchange them, dislodgment, restenosis, biocompatibility, and poor mechanical properties. Sustainable effectiveness, attenuation and prevention of fibrosis, and biocompatibility are key factors for the clinical application of stents to BBS treatment. In addition, loading drugs could show synergistic effects with stents' own performance. We developed a dexamethasone-eluting biodegradable stent (DBS) consisting of a sheath/core structure with outstanding mechanical properties and sustained release of dexamethasone, which maintained its functions in a BBS duct over 12 weeks in a swine model. The insertion of our DBS not only expanded BBS areas but also healed secondary ulcers as a result of the attenuation of fibrosis. After 16 weeks from the insertion, BBS areas were totally improved, and the DBS was degraded and thoroughly disappeared without re-intervention for stent removal. Our DBS would be an effective clinical tool for non-vascular diseases. STATEMENT OF SIGNIFICANCE: This study describes the insertion of a drug-eluting biodegradable stent (DBS) into the bile duct. The sheath/core structure of DBS confers substantial durability and a sustained drug release profile. Drug released from the DBS exhibited anti-fibrotic effects without inflammatory responses in both in vitro and in vivo experiments. The DBS maintained its function over 12 weeks after insertion into the common bile duct, expanding benign biliary stricture (BBS) and reducing inflammation to heal secondary ulcers in a swine BBS model. After 16 weeks from the DBS insertion, the DBS thoroughly disappeared without re-intervention for stent removal, resulting in totally improved BBS areas. Our findings not only spotlight the understanding of the sheath/core structure of the biodegradable stent, but also pave the way for the further application for non-vascular diseases.

Mesenchymal stem cell-derived secretomes-enriched alginate/ extracellular matrix hydrogel patch accelerates skin wound healing.

BACKGROUND: The secretomes of mesenchymal stem cells (MSCs) have great therapeutic potential and thereby their efficient delivery into the target site is of particular interest. Here, we propose a new strategy of hMSCs-derived secretomes delivery for advanced wound healing upon harnessing the working principle of extracellular matrix (ECM)-growth factors interaction in vivo. METHODS: We prepared an alginate hydrogel based wound patch, where it contains both human MSC-derived secretomes and ECM. The ECM was obtained from the decellularization of in vitro cultured human lung fibroblasts. The alginate solution was blended with ECM suspension, crosslinked, air-dried, then rehydrated with the secretomes contained in the concentrated conditioned media (CCM) as a highly saturated form of conditioned media (CM). We tested four different groups, with or without the ECM to investigate not only the role of ECM but the therapeutic effect of secretomes. RESULTS: The secretomes reserved many, diverse bioactive factors, such as VEGF, HGF, IGFBPs, IL-6, and IL-8. Alginate/ECM/CCM (AEC) patch could hold significantly larger amount of secretomes and release them longer than the other groups. Our AEC patch was the most effective in stimulating not only cell migration and proliferation but the collagen synthesis of dermal fibroblasts in vitro. Moreover, the AEC patch-treated full-thickness skin wounds disclosed significantly better wound healing indications: cell recruitment, neovascularization, epidermis thickness, keratinocyte migration, and mature collagen deposition, as assessed via histology (H&E, Herovici staining) and immunofluorescence, respectively. In particular, our AEC patch enabled a phenotype shift of myofibroblast into fibroblast over time and led to mature blood vessel formation at 14 day. CONCLUSIONS: We believe that ECM certainly contributed to generate a secretomes-enriched milieu via ECM-secretomes interactions and thereby such secretomes could be delivered more efficiently, exerting significant therapeutic impact either individually or collectively during wound healing process.

Epigallocatechin-3-Gallate as a Novel Vaccine Adjuvant.

Vaccine adjuvants from natural resources have been utilized for enhancing vaccine efficacy against infectious diseases. This study examined the potential use of catechins, polyphenolic materials derived from green tea, as adjuvants for subunit and inactivated vaccines. Previously, catechins have been documented to have irreversible virucidal function, with the possible applicability in the inactivated viral vaccine platform. In a mouse model, the coadministration of epigallocatechin-3-gallate (EGCG) with influenza hemagglutinin (HA) antigens induced high levels of neutralizing antibodies, comparable to that induced by alum, providing complete protection against the lethal challenge. Adjuvant effects were observed for all types of HA antigens, including recombinant full-length HA and HA1 globular domain, and egg-derived inactivated split influenza vaccines. The combination of alum and EGCG further increased neutralizing (NT) antibody titers with the corresponding hemagglutination inhibition (HI) titers, demonstrating a dose-sparing effect. Remarkably, EGCG induced immunoglobulin isotype switching from IgG1 to IgG2a (approximately >64-700 fold increase), exerting a more balanced TH1/TH2 response compared to alum. The upregulation of IgG2a correlated with significant enhancement of antibody-dependent cellular cytotoxicity (ADCC) function (approximately 14 fold increase), providing a potent effector-mediated protection in addition to NT and HI. As the first report on a novel class of vaccine adjuvants with built-in virucidal activities, the results of this study will help improve the efficacy and safety of vaccines for pandemic preparedness.

Comparative Safety Profiles of Sedatives Commonly Used in Clinical Practice: A 10-Year Nationwide Pharmacovigilance Study in Korea.

This study aims to compare the prevalence and seriousness of adverse events (AEs) among sedatives used in critically ill patients or patients undergoing invasive procedures and to identify factors associated with serious AEs. Retrospective cross-sectional analysis of sedative-related AEs voluntarily reported to the Korea Adverse Event Reporting System from 2008 to 2017 was performed. All AEs were grouped using preferred terms and System Organ Classes per the World Health Organization-Adverse Reaction Terminology. Logistic regression was performed to identify factors associated with serious events. Among 95,188 AEs, including 3132 (3.3%) serious events, the most common etiologic sedative was fentanyl (58.8%), followed by pethidine (25.9%). Gastrointestinal disorders (54.2%) were the most frequent AEs. The most common serious AE was heart rate/rhythm disorders (33.1%). Serious AEs were significantly associated with male sex; pediatrics; etiologic sedative with etomidate at the highest risk, followed by dexmedetomidine, ketamine, and propofol; polypharmacy; combined sedative use; and concurrent use of corticosteroids, aspirin, neuromuscular blockers, and antihistamines (reporting odds ratio > 1, p < 0.001 for all). Sedative-induced AEs are most frequently reported with fentanyl, primarily manifesting as gastrointestinal disorders. Etomidate is associated with the highest risk of serious AEs, with the most common serious events being heart rate/rhythm disorders.

Influence of shoulder coverage difference of abutment on stress distribution and screw stability in tissue-level internal connection implants: A finite element analysis and in vitro study.

STATEMENT OF PROBLEM: Tissue-level internal connection implants are widely used, but the difference in abutment screw stability because of the shoulder coverage formed by the contact between the shoulder of the implant collar and the abutment remains unclear. PURPOSE: The purpose of this finite element analysis (FEA) and in vitro study was to investigate stress distribution and abutment screw stability as per the difference in shoulder coverage of the abutment in tissue-level internal connection implants. MATERIAL AND METHODS: Abutments were designed in 3 groups as per the shoulder coverage of the implant collar, yielding complete coverage (complete group), half coverage (half group), no coverage (no group) groups. In the FEA, a tightening torque of 30.0 Ncm was applied to the abutment screw, a force of 250 N was applied to the crown at a 30-degree angle, and the von Mises stresses and the stress distribution patterns were evaluated. In the in vitro study, the groups were tested (n=12). A total of 200 000 cyclic loads were applied at 250 N, 14 Hz, and at a 30-degree angle. Removal torque values and scanning electron microscopy (SEM) images were assessed. Removal torque values were analyzed by ANOVA and paired t tests. RESULTS: The maximum von Mises stress of the abutment screw was the lowest in the complete group, slightly higher in the half group, and highest in the no group. High stresses were concentrated in 1 location in the implant abutment connection area of the no group. The removal torque values after loading were significantly lower in the no group than in the complete group (P=.047). The SEM images revealed concentrated structural loss and wear in 1 location of the no group. CONCLUSIONS: FEA and in vitro studies confirmed that the shoulder coverage of the abutment in the tissue-level internal connection implant helped improve screw stability. Cyclic loading reduced the removal torque of the abutment screw.

Regulation of mRNA export through API5 and nuclear FGF2 interaction.

API5 (APoptosis Inhibitor 5) and nuclear FGF2 (Fibroblast Growth Factor 2) are upregulated in various human cancers and are correlated with poor prognosis. Although their physical interaction has been identified, the function related to the resulting complex is unknown. Here, we determined the crystal structure of the API5-FGF2 complex and identified critical residues driving the protein interaction. These findings provided a structural basis for the nuclear localization of the FGF2 isoform lacking a canonical nuclear localization signal and identified a cryptic nuclear localization sequence in FGF2. The interaction between API5 and FGF2 was important for mRNA nuclear export through both the TREX and eIF4E/LRPPRC mRNA export complexes, thus regulating the export of bulk mRNA and specific mRNAs containing eIF4E sensitivity elements, such as c-MYC and cyclin D1. These data show the newly identified molecular function of API5 and nuclear FGF2, and provide a clue to understanding the dynamic regulation of mRNA export.

Effects of the plane of nutrition during the latter grower and entire finisher phases on grow-finish pig performance in summer.

The present study was undertaken to investigate the effects of the plane of nutrition (PN) for growing-finishing pigs on growth performance and meat quality in summer. One hundred and two barrows and 102 gilts weighing approximately 44 kg were placed on a high-, medium-, or low-plane grower diet (HPG, MPG, or LPG) with ME and lysine concentrations ranging from 3.33 to 3.40 Mcal/kg and 0.93% to 1.15%, respectively, for 29 days in 6 replicates (pens) in total. Pigs from each grower pen were divided into two finisher pens and provided with a high-plane finisher diet (HPF) containing 3.40 Mcal ME and 9.5 g lysine/kg and a low-plane finisher diet (LPF; 3.25 Mcal ME and 8 g lysine/kg), respectively, up to approximately 110 kg, and slaughtered. Growth performance of the pigs, including average daily gain (ADG), average daily feed intake (ADFI), and gain:feed ratio, was not influenced by the grower-phase PN during any of the grower phase, a 31-d finisher phase I, and ensuing phase II. However, both the ADG and gain:feed ratio were greater (p < 0.05) for the HPF group than for the LPF group during the finisher phase I (748 vs. 653 g with SEM = 13 g and 0.333 vs. 0.299 with SEM = 0.008, respectively). The ADG, but not gain:feed ratio, was greater for the HPF group vs. LPF during the finisher phase II (673 vs. 623 g with SEM = 15 g for ADG and 0.322 vs. 0.323 with SEM = 0.005 for the gain:feed ratio). The carcass backfat thickness (BFT) was greater for the LPF group vs. HPF within the pigs which had been placed on LPG during the grower phase, but not within the pigs from the HPG or MPG group. Physicochemical characteristics of the longissimus dorsi muscle (LM) and sensory quality attributes of fresh and cooked LM were not influenced by either the grower-phase or finisher-phase PN. In conclusion, high PN is necessary for finishing pigs during the hot season to minimize the reduced rate of weight gain and also to prevent the increase of BFT that could result from low PN.

Enamel wear and aging of translucent zirconias: In vitro and clinical studies.

STATEMENT OF PROBLEM: Zirconia is a widely used restorative material. However, phase transformation on clinical application of zirconia has not yet been studied. PURPOSE: The purpose of this study was to evaluate the wear, surface roughness, and aging associated with polished translucent zirconia in both in vitro and clinical experiments. MATERIAL AND METHODS: In vitro experiments were performed with Rainbow and Katana zirconia blocks and natural tooth enamel as the control. They were subjected to 100 000 loading cycles with a maxillary premolar antagonist. All specimens were analyzed for wear, and the zirconia specimens were evaluated for surface roughness and monoclinic phase (m-phase) transformation by X-ray diffractometry before and after cyclic loading. The clinical study included participants who required single-crown implant-supported restorations replacing the first or second molar. The participants received Rainbow or Katana zirconia prostheses (n=15, each). For wear analysis, impressions of each prosthesis, antagonist, and adjacent tooth were made at 1 week and 6 months after crown delivery. The occlusal relationship of the crowns in maximum intercuspation was evaluated by using the T-Scan 8 occlusal diagnostic system. The degree of transformation of zirconia to the m-phase was measured by using X-ray diffractometry of the crowns after 6 months of use. RESULTS: Zirconia induced significantly greater enamel wear than the natural tooth control. Katana specimens exhibited significantly greater wear and surface roughness than the Rainbow specimens. The degrees of antagonistic wear and zirconia phase transformation in the clinical experiment were significantly greater than those in the in vitro experiment. The Katana groups showed significantly higher m-phase levels than the Rainbow groups. CONCLUSIONS: Phase transformation of zirconia occurs within 6 months of clinical use, and the wear and degrees of phase transformation varied according to the zirconia product used.

The reproducibility and validity verification for body composition measuring devices using bioelectrical impedance analysis in Korean adults.

The purpose of this study is to examine the validity and reproducibility of impedance body fat measurement devices measuring the body composition of Korean male and female adults using three bioelectrical impedance analyzers. We compared two methods for evaluating body composition: dual energy X-ray absorptiometry (DEXA), and bioelectrical impedance analysis (BIA). Subjects were 200 healthy adult Korean males and females whose mean±standard deviation (range) age, standing height, body weight, and body mass index (BMI) were 44.1±14.5 years, 172.8±7.4 cm, 76.0±12.8 kg, and 25.4±3.3 kg/m2, and 44.5±14.7 years, 158.7±5.8 cm, 58.3±8.3 kg, and 23.2±3.0 kg/m2, respectively. As a result, first of all, the reproducibility of the bioelectrical impedance analyzer had very high coefficients at r=0.998, r=0.997 between men and women, respectively. The correlation coefficients among three comparisons for lean body mass (LBM) were provided the following coefficients: r=0.951 for DEXA vs. ACCUNIQ BC720, r=0.950 for DEXA vs. ACCUNIQ BC360, and r=0.946 for DEXA vs. ACCUNIQ BC380 in men. In the results for women, they also had the very high following coefficients: r=0.956 for DEXA vs. ACCUNIQ BC720, r=0.946 for DEXA vs. ACCUNIQ BC360, and r=0.957 for DEXA vs. ACCUNIQ BC380 in LBM. In conclusion, this research showed a higher correlation in terms of accuracy compared to existing BIA-based body composition measurement techniques, and the accuracy of LBM was improved with high correlation coefficients through the algorithm that was improved using the multifrequency BIA method in the ACCUNIQ BC products.

Association between Cigarette Smoking Frequency and Health Factors among Korean Adults.

BACKGROUND: Recently, there has been a trend that cigarette smoking rate in Asian and Africa adults has increased while the age group to start smoking has decreased gradually. This study aimed to investigate the relationships between lifetime smoking and hypertension, diabetes, obesity, waist measure, fasting blood pressure and food consumption, in order to look into health status depending on smoking status in Koreans. METHODS: Totally, 1075 men and 697 women with no disease participated in this study, in which one-way ANOVA was conducted by using SPSS version 18.0 for statistical process. The level of statistical significance was 0.05. RESULTS: As a result of analysis on relationship between lifetime smoking and hypertension, obesity and diabetes, statistically significant differences were revealed.Lifetime smoking was found to be significantly associated with increased waist measure, higher level of fasting blood sugar, and more ingestion of nutrients (carbohydrate, fat, and protein). CONCLUSION: Increased amount of lifetime cigarette smoking was shown to negatively influence various health factors, which might become to be a drive to cause diseases. Therefore, method to improve health factors must be sought for via education and campaign to control an amount of cigarette smoking in Korean adults.

Correction: Crystal Structure of Human Myotubularin-Related Protein 1 Provides Insight into the Structural Basis of Substrate Specificity.

[This corrects the article DOI: 10.1371/journal.pone.0152611.].

Harnessing an RNA-mediated chaperone for the assembly of influenza hemagglutinin in an immunologically relevant conformation.

A novel protein-folding function of RNA has been recognized, which can outperform previously known molecular chaperone proteins. The RNA as a molecular chaperone (chaperna) activity is intrinsic to some ribozymes and is operational during viral infections. Our purpose was to test whether influenza hemagglutinin (HA) can be assembled in a soluble, trimeric, and immunologically activating conformation by means of an RNA molecular chaperone (chaperna) activity. An RNA-interacting domain (RID) from the host being immunized was selected as a docking tag for RNA binding, which served as a transducer for the chaperna function for de novo folding and trimeric assembly of RID-HA1. Mutations that affect tRNA binding greatly increased the soluble aggregation defective in trimer assembly, suggesting that RNA interaction critically controls the kinetic network in the folding/assembly pathway. Immunization of mice resulted in strong hemagglutination inhibition and high titers of a neutralizing antibody, providing sterile protection against a lethal challenge and confirming the immunologically relevant HA conformation. The results may be translated into a rapid response to a new influenza pandemic. The harnessing of the novel chaperna described herein with immunologically tailored antigen-folding functions should serve as a robust prophylactic and diagnostic tool for viral infections.-Yang, S. W., Jang, Y. H., Kwon, S. B., Lee, Y. J., Chae, W., Byun, Y. H., Kim, P., Park, C., Lee, Y. J., Kim, C. K., Kim, Y. S., Choi, S. I., Seong, B. L. Harnessing an RNA-mediated chaperone for the assembly of influenza hemagglutinin in an immunologically relevant conformation.

Correlation between sagittal condylar guidance angles obtained using radiographic and protrusive occlusal record methods.

PURPOSE: This study compared the SCGAs measured in three types of radiographic images (panoramic, CBCT panoramic-section, and CBCT cross-section images) with values measured using the protrusive occlusal record. MATERIALS AND METHODS: SCGAs were measured in 20 patients on a semi-adjustable articulator using the protrusive interocclusal record. Panoramic and CBCT images were obtained. SCGAs were measured on CBCT images in panoramic and cross sections. In all of the radiographic images, SCGAs were measured using the Frankfort horizontal reference line and the mean curvature line. The most-superior and most-inferior points of the curvatures were identified to determine the mean curvature line. Each measurement was performed twice by two operators independently. The data were analyzed by the t-test, Pearson's correlation test, and Cronbach's α using SPSS (α=.05). RESULTS: The mean right and left SCGAs were as follows: protrusive occlusal record (30.1 and 30.2 degrees, respectively), panoramic (38.9 and 38.7 degrees), CBCT panoramic sections (35.4 and 36.8 degrees), and CBCT cross sections (35.3 and 36.1 degrees). The SCGAs differed significantly among the groups. The Pearson coefficients for the correlations with the protrusive occlusal record measurements on the left and right sides were as follows: panoramic (0.834 and 0.791, respectively), CBCT panoramic-section (0.918 and 0.837), and CBCT cross-section (0.918 and 0.845) images. CONCLUSION: Strong correlations were found between SCGAs obtained using radiographic images and the protrusive occlusal record.

Crystal Structure of Human Myotubularin-Related Protein 1 Provides Insight into the Structural Basis of Substrate Specificity.

Myotubularin-related protein 1 (MTMR1) is a phosphatase that belongs to the tyrosine/dual-specificity phosphatase superfamily. MTMR1 has been shown to use phosphatidylinositol 3-monophosphate (PI(3)P) and/or phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2) as substrates. Here, we determined the crystal structure of human MTMR1. The refined model consists of the Pleckstrin homology (PH)-GRAM and phosphatase (PTP) domains. The overall structure was highly similar to the previously reported MTMR2 structure. Interestingly, two phosphate molecules were coordinated by strictly conserved residues located in the C(X)5R motif of the active site. Additionally, our biochemical studies confirmed the substrate specificity of MTMR1 for PI(3)P and PI(3,5)P2 over other phosphatidylinositol phosphates. Our structural and enzymatic analyses provide insight into the catalytic mechanism and biochemical properties of MTMR1.

Genetic analysis of attenuation markers of cold-adapted X-31 influenza live vaccine donor strain.

Cold-adapted live attenuated influenza vaccines (CAIVs) have been considered as a safe prophylactic measure to prevent influenza virus infections. The safety of a CAIV depends largely on genetic markers that confer specific attenuation phenotypes. Previous studies with other CAIVs reported that polymerase genes were primarily responsible for the attenuation. Here, we analyzed the genetic mutations and their phenotypic contribution in the X-31 ca strain, a recently developed alternative CAIV donor strain. During the cold-adaptation of its parental X-31 virus, various numbers of sequence changes were accumulated in all six internal genes. Phenotypic analysis with single-gene and multiple-gene reassortant viruses suggests that NP gene makes the largest contribution to the cold-adapted (ca) and temperature-sensitive (ts) characters, while the remaining other internal genes also impart attenuation characters with varying degrees. A balanced contribution of all internal genes to the attenuation suggests that X-31 ca could serve as an ideal master donor strain for CAIVs preventing influenza epidemics and pandemics.

Crystallization and preliminary X-ray crystallographic analysis of human myotubularin-related protein 1.

Myotubularin-related protein 1 is a phosphatase that dephosphorylates phospholipids such as phosphatidylinositol 3-phosphate or phosphatidylinositol 3,5-bisphosphate. In this study, human MTMR1 was overexpressed in Escherichia coli, purified and crystallized at 277 K using polyethylene glycol 20,000 as a precipitant. Diffraction data were collected to 2.0 Šresolution using synchrotron radiation. The crystals belonged to space group P1, with unit-cell parameters a = 67.219, b = 96.587, c = 97.581 Å, α = 87.597, ß = 86.072, γ = 77.327°. Assuming the presence of four molecules in the asymmetric unit, the calculated Matthews coefficient value was 2.61 Å(3) Da(-1) and the corresponding solvent content was 52.9%.

Assessment of professional baseball players aerobic exercise performance depending on their positions.

This study reports the average and SD of professional baseball players' cardiorespiratory endurance, maximum oxygen consumption, oxygen consumption during anaerobic threshold, maximum oxygen consumption of anaerobic threshold %, maximum heart rate, and heart rate of anaerobic threshold. We also report the comparison between pitchers and fielders. Considering the total number of results, percentile was used and results were classified into 5 grades. One professional baseball players' organization with more than 14 years of experience participated in this study. First, we observed that the average pitchers' V[Combining Dot Above]O2max was 53.64 ml·kg·min, whereas the average fielders' was 52.30 ml·kg·min. These values were lower than other sports players. Second, in case of the V[Combining Dot Above]O2AT, pitchers showed 39.35 ml·kg·min and fielders showed 39.96 ml·kg·min. %V[Combining Dot Above]O2AT showed a significant difference of 71.13% between pitchers and fielders-pitchers, whereas fielders showed 73.89% (p < 0.01). Third, maximal heart rates were measured at 188.69 b·min (pitchers) and 187.79 b·min (fielders). These were lower than college baseball players and higher than other sports players. In conclusion, both professional baseball pitchers and fielders should be aware of the necessity of systematic cardiorespiratory endurance data analysis. Moreover, baseball teams, athletic trainers, and coaches should also be aware of the importance of cardiorespiratory endurance.