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Dried Blood Spots (DBS) revolutionize therapeutic drug monitoring using LC-MS for the precise quantification of cardiovascular drugs (CDs), enabling personalized treatment adapted to patient-specific pharmacokinetics with minimal invasiveness. This study aims to achieve simultaneous quantification of eight CDs in DBS, overcoming physicochemical challenges. A two-step protein precipitation method was used for simple and precise sample preparation. The drugs were analyzed using LC-MS/MS in ESI positive-ion mode, showing high sensitivity and linearity, with a correlation coefficient (r 2 ) exceeding 0.999, after being separated on a reversed-phase chromatography by gradient elution of DW-acetonitrile containing 0.1 % formic acid + 2 mM ammonium formate. The validation results indicate good selectivity, with no observed matrix effect and carry-over. The intra- and inter-day accuracy and precision were within 6 % for most drugs, except for digoxin and deslanoside at low therapeutic levels where the variation was within 20 %. Stability tests confirmed suitable DBS handling and storage conditions, indicating drug stability for at least 30 days at room temperature. The analysis of whole spot has demonstrated remarkable precision and reliability in all target drugs. The analysis of 3 mm internal diameter discs, punched in and out of DBS, presumed to contain 3 µL of blood, showed acceptable accuracy for most drugs, with less polar drugs like digoxin and deslanoside showing lower accuracy, indicating a need for further correction due to non-uniform drug distribution. Consequently, the developed LC-MS/MS method enables the quantification of multiple CDs in a single DBS analysis, while suggesting the potential for accuracy-based analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

This study aims to establish an LC-MS/MS method to simultaneously analyze 11 antiepileptic drugs with a particular focus on maintaining accuracy while reducing the number of isotope-labeled internal standards employed for cost-effectiveness. By applying a water/acetonitrile gradient elution containing 0.1 % formic acid and 2 mM ammonium formate as the mobile phase, optimal sensitivity for the target drugs could be obtained in positive ESI mode in LC-MS/MS. After optimizing various extraction techniques, extraction with 70 % acetonitrile was selected as it provided good recoveries (>93 %) for all targets without matrix effects. Accuracies within 3 % were achieved from the combination of six internal standards, while accuracies of 5 % and 10 % were obtained by reducing the number of internal standards to four and two, respectively, for more economical analysis. The accuracy of the established method was maintained in hyperglycemia, hyperlipidemia, and hyperalbuminemia sera, suggesting that it can be successfully applied to individual serum samples with various properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Study Objective: To investigate the analgesic efficacy of erector spinae plane block (ESPB) in major gynecologic surgery, expressed as cumulative opioid consumption 24 h after surgery., Design: A single-center, patient-assessor blinded, randomized controlled study., Setting: Samsung medical center (tertiary university hospital), between February 2022 to January 2023., Patients: Eighty-eight females undergoing major surgery with long midline incision for gynecologic malignancy., Interventions: Patients were randomly assigned to receive standard systemic analgesia (Control group) or ESPB (ESPB group). ESPB was performed bilaterally at the level of the 9th thoracic vertebra with a mixture of 20 mL of 0.5% ropivacaine and 100 μg of epinephrine., Measurements: The primary outcome was cumulative opioid consumption at 24 h postoperatively. Secondary outcomes included opioid consumption and pain severity during the 72 h after surgery. The variables regarding postoperative recovery and patient-centered outcomes were compared., Main Results: The mean cumulative opioid consumption 24 h after surgery was 35.8 mg in the ESPB group, which was not significantly different from 41.4 mg in the control group (mean difference, 5.5 mg; 95% CI -1.7 to 12.8 mg; P = 0.128). However, patient satisfaction regarding analgesia was significantly higher in the ESPB group compared with the control group at 24 h postoperative (median difference, -1; 95% CI -3 to 0; P = 0.038). There were no significant differences in the variables associated with postoperative recovery., Conclusion: ESPB did not reduce opioid consumption during the 24 h postoperative but attenuated pain intensity during the early period after surgery., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflicts of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Lincomycin (LCM) is an antibiotic used to treat severe bacterial infections in livestock and companion animals. In this study, we aimed to investigate the oral bioavailability of LCM with PK data after IV and PO administration and to compare differences in drug residue patterns in eggs. To ensure food safety, an additional study on egg residue was conducted using 3 different commercial LCM drugs. For bioavailability study, laying hens were divided into oral and intravenous (n = 8/group) groups and received single dose (10 mg/kg) of LCM. The limits of quantification for LCM were 0.729 μg/mL and 0.009 mg/kg in plasma and eggs, respectively. The oral group exhibited a significantly lower average serum drug concentration than the IV group, with a bioavailability of 2.6%. Furthermore, the egg residue profiles confirmed reduced systemic drug exposure after oral administration. For the commercial LCM drug egg residue experiment, laying hens were divided into low- and high-dose groups (n = 12/group) for each drug and treated with the recommended dosage and administration method for each respective drug. The eggs were collected and analyzed until 14 d after the last drug treatment. Despite differences in the LCM content and formulation among commercial drugs, all the tested commercial drugs showed average concentrations below the MRL in eggs within approximately 3 d after the last drug treatment. In this study, we have confirmed that LCM has a low oral absorption rate in laying hens, and this was consistent with the findings from the egg residue profiles. Further studies are requested to elucidate the exact reasons for evidently low oral drug absorption in laying hens., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

We describe the simultaneous quantification of six antiviral drugs in serum based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The target drugs-hydroxychloroquine, chloroquine, favipiravir, umifenovir, ritonavir, and lopinavir-were extracted and purified from serum with 75 % v/v methanol as the precipitant reagent. The six analytes were clearly separated within 15 min using gradient elution and mixed-mode stationary phase. The measurement accuracy and precision were assured by adopting isotopes as internal standards. The optimized measurement procedure was strictly validated in linearity, sensitivity, accuracy, and precision. To confirm the robustness of the method in matrix, the method was additionally applied to various types of serum, namely hyperlipidemic and hyperglycemic serum. The method was then applied to assess the stability of the drugs in serum in order to set sample handling and storage guides for laboratory testing. Lastly, the method was implemented in different LC-MS systems to confirm its applicability across similar equipment commonly used in clinical testing laboratories. The overall results show that the optimized protocol is suitable for the accurate, simultaneous quantification of the six antiviral drugs in serum, and it is anticipated to satisfactorily serve as a reference protocol for the analysis of a wide range of other antiviral drugs for drug monitoring with various purposes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

The levamisole maximum residue limit for edible fat, kidney, and muscle of chickens is 0.01 mg/kg. However, no maximum residue limit has been established for eggs. In the present study, the pharmacokinetic profile and levamisole residue in the eggs from laying hens were investigated using ultra-performance liquid chromatography-tandem mass spectrometry. A single dose of levamisole (30 mg/kg) was administered via the intramuscular or oral route, and an additional egg residue study was performed with 300 or 600 mg/kg commercial LEV drug (30 or 60 mg/kg as levamisole) orally. The limit of quantification was 0.0056 μg/mL and 0.0015 mg/kg for plasma and eggs, respectively. The plasma concentration was below the limit of quantification 10 and 12 h after intramuscular and oral administration, respectively. The half-life of the absorption phase was comparable between the intramuscular and oral routes, which was approximately 1 h, and the mean maximum concentration value was significantly higher in intramuscular (2.29 ± 0.30 μg/mL) than in oral (1.45 ± 0.38 μg/mL) route. The relative oral bioavailability after intramuscular administration was 92.3%. In the egg residue study, dose-dependent area under concentration and maximum concentration were observed after single oral administration of 30 and 60 mg/kg egg residue, and the calculated withdrawal period for both 30 and 60 mg/kg groups based on the positive list system standard (0.01 mg/kg) was 7 d after the treatment., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Therapeutic drug monitoring (TDM) of cardiovascular drugs is essential to improve treatment efficacy and minimize toxicity because of the usage of multiple drugs with a very limited therapeutic range and the high pharmacokinetic variation in patients. We developed and validated a reliable and economical liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the determination of seven cardiovascular drugs-procainamide, lidocaine, quinidine, deslanoside, digoxin, atorvastatin, and digitoxin-for clinical usage. Serum samples were prepared by simple protein precipitation with an organic solvent consisting of acetonitrile and methanol (2:1 v/v) and analyzed under optimized LC-MS/MS conditions. The chromatographic separations were accomplished within 15 min on a reversed-phase C18 column with a gradient elution of aqueous solvent and acetonitrile while maintaining 0.1 (v/v) % formic acid and 2 mM ammonium formate. The optimized MS/MS conditions in ESI-positive mode offered sufficient sensitivity for the seven cardiovascular drugs (LOQs between 0.5 and 1 ng/mL). This method was fully validated including linearity, selectivity, accuracy, precision, carry-over, and matrix effects. Additionally, stability under several conditions was tested to determine how to handle the standard solutions and serum samples. The seven cardiovascular drugs, simultaneously, were precisely and accurately analyzed in intra- and inter-day assays (RSD < 6 % and recovery between 96.3 and 102.8 %) using only two isotope-labeled internal standards (lidocaine-(diethyl-d10) and digoxin-21, 21, 22-d3). The presented method also showed good accuracy in analyzing the seven drugs in hyperlipidemia, hyperalbuminemia, and hyperglycemia serum, allowing it to be recommended as a common and routine analysis method for cardiovascular drugs in clinical practice., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Bisphenol F (BPF, 4,4'-methylenediphenol) has recently been selected as an alternative to bisphenol A (BPA), which is used in the manufacturing of polycarbonates and epoxy resins. This study aimed to investigate the general, and reproductive/developmental effects of BPF. Therefore, BPF at dose levels of 0, 1, 5, 20, and 100 mg/kg/day was administered daily by oral gavage to Sprague-Dawley rats during the pre-mating, mating, gestation, and early lactation periods, and reproductive and developmental toxicities including general systemic toxicities were investigated. A decrease in body weight and food consumption was observed in the female rats treated with BPF at 20 and 100 mg/kg/day during the pre-mating and gestation periods. Additionally, gamma glutamyl transpeptidase levels were increased in the female rats administered 100 mg/kg/day. At 100 mg/kg/day, ovarian weight decreased and vaginal mucification increased according to a necropsy and histopathological examination, respectively. Moreover, the number of implantation sites and litter size decreased at 100 mg/kg/day. However, no significant BPF-related changes were observed in the male rats. Based on the results of this study, the no-observed-adverse-effect levels (NOAELs) of BPF for general systemic and reproductive effects were 5 and 20 mg/kg/day, respectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The incidence of thyroid cancer (TC) has increased considerably in the last few decades. Environmental factors, including plasticizers, are recognized as potential risks leading to thyroid cancer in humans. In this study, we used a transcriptome-metabolome-wide association study to find the unidentified carcinogenic mechanism of di-2-ethylhexyl phthalate (DEHP) in thyroid and biomarkers for non-invasive diagnosis. Rats were treated with different doses of DEHP (0, 0.3, 3, 30, 150 mg DEHP/kg bw/day) for 13 weeks. Then, the thyroids were processed for Ki67 staining and RNA-seq. Also, 17-h urine samples were collected for high-resolution metabolomics analysis. After a high dose of DEHP exposure, the terminal body weights and the thyroid and parathyroid glands weights were not altered. However, the liver weights and numbers of Ki67-positive cells were increased. Further, multivariate statistical analysis revealed that metabolic shifts were considerably altered above 30 mg DEHP/kg bw/day. In RNA-seq analysis, some cancer-related genes were altered, including 18 upregulated and 9 downregulated transcripts. These cancer transcripts and whole metabolome data were integrated to uncover thyroid cancer-related metabolic pathways, which revealed that cancer-related transcripts had a network structure linked to eicosanoids such as leukotriene D4 and prostaglandin. In brief, our study demonstrated that DEHP can induce thyroid hyperplasia through the eicosanoid-associated pathway, providing further insight into the mechanism of DEHP-associated thyroid cancer., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: The appropriate size of GlideScope® blade for tracheal intubation in neonates and premature infants has not been established. We evaluated the impact of the size of the GlideScope® blade on the time taken for intubation in infants weighing 2.5-3.6 kg., Methods: Sixty infants weighing 2.5-3.6 kg were randomly assigned to use of the size 1 blade (n=30) or the size 2 blade (n=30). The primary outcome was the time taken to intubate. Components related to the laryngoscopic view which could affect the duration of the intubating process were also analysed., Results: The time required for tracheal intubation was shorter with the size 2 blade than with the size 1 blade (16 [14-20] s vs 22 [18-25] s, P=0.002; median difference=-5; 95% confidence interval, -7 to -2). The rate at which the tip of the tracheal tube was located at the centre of the laryngeal inlet was higher with the size 2 blade than with the size 1 blade (83% vs 40%, P<0.001). Correlation analysis indicated that the time required to find the tip of the tube was related to how far the lower border of the arytenoid cartilages was located from the mid-horizontal line of the monitor (r=0.28, P=0.033)., Conclusion: Use of the size 2 blade significantly reduced the time required to intubate the trachea, compared with the size 1 GlideScope® blade in infants., Clinical Trial Registration: KCT 0003867., (Copyright © 2022 British Journal of Anaesthesia. Published by Elsevier Ltd. All rights reserved.)

Sore throat lozenges, which are over-the-counter drugs, contain 2,4-dichlorobenzyl alcohol (DCBA) as the primary ingredient. However, comprehensive data on the prenatal developmental toxicity of DCBA is limited. Therefore, this study was conducted to determine the effects of DCBA on pregnant rats and prenatal development. Sprague-Dawley rats were administered different doses of DCBA (0, 25, 100, 400, and 800 mg/kg/day) daily via an oral gavage from gestation day (GD) 6-19. Thereafter, all the live dams were sacrificed on GD 20, and caesarean sections were conducted. Live fetuses and their placenta were weighed and then examined for external, visceral, and skeletal malformations and variations. Based on the results obtained, dams at 800 mg/kg/day showed systemic toxicities, including a decrease in body weight and food consumption, and liver changes. Additionally, this treatment induced decreases in fetal and placental weights, as well as the increased incidence of retarded ossifications and full supernumery rib, and the decreased number of ossification centers. Therefore, based on these findings, the no-observed-adverse-effect level of DCBA was determined to be 400 mg/kg/day for dams and prenatal development., (Copyright © 2022. Published by Elsevier Inc.)

Alopecia is defined as hair loss in a part of the head due to various causes, such as drugs, stress and autoimmune disorders. Various therapeutic agents have been suggested depending on the cause of the condition and patient sex, and age. Minoxidil (MXD) is commonly used topically to treat alopecia, but its low absorption rate limits widespread use. To overcome the low absorption, we suggest microneedles (MNs) as controlled drug delivery systems that release MXD. We used hyaluronic acid (HA) to construct MN, as it is biocompatible and safe. We examined the effect of HA on the hair dermal papilla (HDP) cells that control the development of hair follicles. HA enhanced proliferation, migration, and aggregation of HDP cell by increasing cell-cell adhesion and decreasing cell substratum. These effects were mediated by the cluster of differentiation (CD)-44 and phosphorylation of serine‑threonine kinase (Akt). In chemotherapy-induced alopecia mice, topical application of HA tended to decrease chemotherapy-induced hair loss. Although the amount of MXD administered by HA-MNs was 10% of topical treatment, the MXD-containing HA-MNs (MXD-HA-MNs) showed better effects on the growth of hair than topical application of MXD. In summary, our results demonstrated that HA reduces hair loss in alopecia mice, and that delivery of MXD and HA using MXD-HA-MNs maximizes therapeutic effects and minimize the side effects of MXD for the treatment of alopecia. STATEMENT OF SIGNIFICANCE: (1) Significance, This work reports a new approach for treatment of alopecia using a dissolving microneedle (MN) prepared with hyaluronic acid (HA). The HA provided a better environment for cellular functions in the hair dermal papilla cells. The HA-MNs containing minoxidil (MXD) exhibited a significant reduction of hair loss, although amount of MXD contained in them was only 10% of topically applied MXD., (2) Scientific impact, This is the first report demonstrating the direct anti-alopecia effects of HA administrated in a transdermal route and the feasibility of novel therapeutics using MXD-containing HA-MNs. We believe that our work will excite interdisciplinary readers of Acta Biomaterialia, those who are interested in the natural polymers, drug delivery, and alopecia., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

Serial blood sampling for toxicokinetics is generally conducted in regulatory embryo-fetal development (EFD) studies in rats. EFD studies are designed to detect the potential adverse effects of pharmaceuticals on pregnant females and their fetuses; this information is useful for understanding the relationships between systemic exposure levels and toxicity profiles. However, additional satellite pregnant females are needed for toxicokinetics because comprehensive information regarding the potential impact of serial blood sampling on pregnant females is scarce. Here, in this study, we investigated the potential impact of serial blood sampling in pregnant female rats using a typical EFD study design. Additionally, we investigated the additional endpoints (clinical pathology, organ weights, and histopathology) that were deemed likely to be sensitive to blood sampling. Results indicated that serial blood sampling in pregnant females induced physiological adaptive changes and did not affect the general endpoints in EFD studies. Nevertheless, inclusion of satellite groups in EFD studies may be a more prudent approach considering the physiological changes in pregnant females and potential off-target effects of candidate pharmaceuticals. These results provide background information on the impact of serial blood sampling in pregnant females and will be useful to design the regulatory EFD studies., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Biocides are widely used for their effective antiseptic and disinfectant properties, including polyhexamethylene guanidine phosphate (PHMG-P), which is also used as a biocide as it selectively disrupts bacterial cell membrane. It is used to clean humidifiers commonly used in the dry winter season in South Korea, which exposes people to PHMG-P inhalation. However, comprehensive toxicological data on PHMG-P inhalation exposure, including in pregnant women, and the potential occurrence of lung disease is lacking. Therefore, in this study, we investigated PHMG-P inhalation exposure-induced toxicities in pregnant rats and prenatal development of their conceptus. Pregnant rats were exposed to PHMG-P via inhalation at target concentrations of 0, 0.14, 1.60, and 3.20 mg/m 3 from implantation to nearly parturition (from gestation day 6-20) and then analyzed for relevant abnormalities. Results showed systemic toxicities in the pregnant rats including respiratory function abnormalities, decreased body weight gain, and decreased food consumption at ≥1.60 mg/m 3 . Prenatal development toxicities, including decreased fetal weight with ossification retardations of fetal bones, were observed at 3.20 mg/m 3 . These results will contribute to clarifying the PHMG-P inhalation exposure-induced toxicities during pregnancy and support its risk assessment in humans., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Monosaccharide composition of biological samples can reflect an individual's health status. Monitoring the concentration of individual monosaccharides in human serum requires a technique for the simultaneous analysis of multiple monosaccharide molecules. Furthermore, certified reference materials (CRMs) for overall monosaccharide composition of human serum are required in order to validate the performance of clinical laboratory instruments. In the present study, we present a novel method for the simultaneous analysis of numerous monosaccharide molecules without the need for derivatization or post-column treatment. We utilized ultra-high-performance liquid chromatography (UHPLC)-quadrupole/orbitrap mass spectrometry incorporating a hydrophilic interaction chromatography (HILIC) column. We optimized the precursor ions, product ions, mobile phase composition and gradient program, flow rate, and column temperature. Seven monosaccharides (D-Ribose, L-Arabinose, D-Xylose, D-Fructose, D-Mannose, D-Galactose and D-Glucose) were able to be separated and quantified. We validated the method and the seven molecules showed favorable limits of detection and quantification, recovery rates, carry-over effects, intra- and inter-day accuracy and precision, resolution, and measurement uncertainty. We analyzed human serum samples using the method. To avoid ion suppression and D-d 2 -Glucose peak interference, compounds present at concentrations outside of the calibration range were analyzed from diluted samples. Quantification of serum samples corroborated some previous clinical research, in that increased D-Glucose concentration was associated with increased concentrations of D-Mannose and D-Ribose. We also validated the CRMs, and expect these to have utility as standards for serum monosaccharide profiling, thus contributing to public health., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: IPNB is very rare disease and most previous studies on IPNB were case series with a small number due to low incidence. The aim of this study is to validate previously known clinicopathologic features of intraductal papillary neoplasm of bile duct (IPNB) based on the first largest multicenter cohort., Methods: Among 587 patients previously diagnosed with IPNB and similar diseases from each center in Korea, 387 were included in this study after central pathologic review. We also reviewed all preoperative image data., Results: Of 387 patients, 176 (45.5%) had invasive carcinoma and 21 (6.0%) lymph node metastasis. The 5-year overall survival was 80.9% for all patients, 88.8% for IPNB with mucosal dysplasia, and 70.5% for IPNB with invasive carcinoma. According to the "Jang & Kim's modified anatomical classification," 265 (68.5%) were intrahepatic, 103 (26.6%) extrahepatic, and 16 (4.1%) diffuse type. Multivariate analysis revealed that tumor invasiveness was a unique predictor for survival analysis. (p = 0.047 [hazard ratio = 2.116, 95% confidence interval 1.010-4.433])., Conclusions: This is the first Korean multicenter study on IPNB through central pathologic and radiologic review process. Although IPNB showed good long-term prognosis, relatively aggressive features were also found in invasive carcinoma and extrahepatic/diffuse type., (Copyright © 2019 International Hepato-Pancreato-Biliary Association Inc. Published by Elsevier Ltd. All rights reserved.)

Objective: To identify the association between the use of acetylcholinesterase inhibitors (AChEIs) and risk of osteoporotic fractures in older persons with Alzheimer's disease (AD)., Design, Setting, and Participants: A nested case-control study was conducted using the Korean National Health Insurance Service-National Elderly Cohort database. Patients with AD who were newly diagnosed with osteoporotic fractures were identified as cases. Up to 3 controls were matched with cases according to age, sex, and duration of follow-up., Methods: Participants were considered as exposed to AChEIs if they had been prescribed at least 1 AChEI during a period of 2 years before the index date. A conditional logistic regression was performed to estimate the adjusted odds ratios with 95% confidence intervals for the association between the use of AChEIs and osteoporotic fractures in patients with AD. We also examined the impact of dose, duration of treatment, and timing of exposure on the estimates of the association between the use of AChEIs and risk of osteoporotic fractures., Results: The study cohort comprised 45,006 patients diagnosed with AD, of which 9470 patients, including 2385 cases and 7085 controls, were available for the study. The mean ages (standard deviations) were 78.6 (6.9) years in the cases and 80.0 (6.9) years in the controls. Adjusted odds ratios for the association between the use of AChEIs and osteoporotic fractures in patients with AD was 1.18 (95% confidence interval 1.07-1.31)., Conclusions and Implications: Our data indicated that the use of AChEIs was not associated with a reduced risk of osteoporotic fractures in patients with AD; in contrast, their use was associated with a mild increased risk of osteoporotic fractures. Thus, clinicians should consider the possibility of AChEIs-associated fractures among older persons with AD. Findings of this study will support shared decision making among prescribers, patients, and caregivers., (Copyright © 2019 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.)

We sequenced the complete mitochondrial genome (mitogenome) of the dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae); the sequence of this mitogenome had not been previously reported for this genus and tribe (Coprini). The mitogenome had the trnP-trnT arrangement at the ND4L-ND6 junction, which is found in only two species among the 56 available Scarabaeoidea species, including C. tripartitus. Twelve protein-coding genes (PCGs) had the typical ATT and ATG start codons, whereas COII gene had the GTA start codon, which is unusual in Scarabaeoidea. Phylogenetic analysis of the 32 species within the Scarabaeidae strongly supported the Onthophagini as non-monophyletic group. Coprini, the newly included tribe within Scarabaeinae, which includes C. tripartitus, was positioned at the most basal lineage of all the tribes included in Scarabaeinae, such as Oniticellini, Onthophagini, Onitini, Dichotomiini, Phanaeini, and Eurysternini with the highest nodal supports when amino acid sequences of 13 PCGs were used. However, analyses based on the nucleotide sequences of two RNAs and 13 PCGs placed Coprini as a sister tribe to the group composed of Dichotomiini, Phanaeini, and Eurysternini with lower nodal supports (ML, 55%; BI, 0.53). Although different datasets provided a conflicting result, higher nodal supports were obtained by amino acid sequence-based analyses., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A study of the STEM probe channeling in a heterostructured crystalline bilayer specimens is presented here with a goal to guide STEM-based characterization of multilayer structures. STEM analysis of perovskite BaSnO 3 /LaAlO 3 bilayers is performed and the dominating effects of beam channeling on HAADF- and LAADF-STEM are illustrated. To study the electron beam channeling through BaSnO 3 /LaAlO 3 bilayers, probe intensity depth profiles are calculated, and the effects of probe defocus and atomic column alignment are discussed. Characteristics of the beam channeling are correlated to resulting ADF-STEM images, which is then tested by comparing focal series of plan-view HAADF-STEM images to those recorded experimentally. Additionally, discussions on how to visualize the misfit dislocation network at the BaSnO 3 /LaAlO 3 interface using HAADF- and LAADF-STEM images are provided., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Polyhexamethylene guanidine phosphate (PHMG-P) has effective antimicrobial activity against various microorganisms and has been widely used as a biocide in commercial products. However, its use as a humidifier disinfectant has provoked fatal idiopathic lung disease in South Korea, especially in pregnant or postpartum women and their young children. PHMG-P-related toxicological studies of reproduction and development in experimental animals have not been identified, and thus, we investigated the potential effects of early-stage oral exposure to PHMG-P by assessing its toxicological properties. PHMG-P was repeatedly administered by oral gavage at dose levels of 0, 13, 40 and 120 mg/kg to Sprague-Dawley rats during the pre-mating, mating, gestation and early lactation periods, and then general systemic and reproductive/developmental toxicities were investigated. At 120 mg/kg, PHMG-P-related toxicities including subdued behavior, thin appearance, decreased body weight, decreased food consumption and decreased F1 pup body weight were observed. Based on the results of this study, the no-observed-adverse-effect levels (NOAELs) of PHMG-P for both general systemic effects and development are considered to be 40 mg/kg/day., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A baseline dip caused by the reduction of dissolved oxygen in samples has been a source of trouble in the analysis of major monosaccharides (galactose, glucose, mannose, and fructose) in the high-performance anion-exchange chromatography with pulsed amperometric detection system. This study attempted three different methods to remove the baseline dip from the resulting chromatograms, and among the approaches, sodium sulfite was found to act as the best oxygen scavenger. Clean chromatograms were obtained by adding at least 3 mg/mL sodium sulfite to samples, which removed the baseline dip and improved the accuracy of sugar analysis. Although sodium sulfite does not influence analytical sensitivity, it can cause a reduction of sugar retention; however, retention time can be recovered by washing with 200 mM sodium hydroxide solution. Results demonstrated that sodium sulfite is an effective means either to remove the baseline dip for low concentration analysis under 1 mg/L, or to separate the target sugar from the baseline dip by retention time rearrangement., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A certified reference material (CRM) for the quantification of protein, essential to manage quality control and quality assurance in protein-related works, has been developed. Amino acid analysis with conventional acid hydrolysis and isotope dilution HPLC-MS was used to establish an SI-traceable absolute protein quantification method using recombinant human growth hormone (hGH) as a model protein. The certification method was verified by comparative studies between 1) different methods of protein quantification based on microwave-assisted hydrolysis, and 2) different labs as part of the Asian Collaboration on Reference Material project with Japan, China, and Korea. Certification, evaluation of measurement uncertainty, homogeneity testing, and stability testing were carried out, after which the candidate CRM for hGH quantification was successfully certified with excellent agreement within the certified value in the two comparative studies. Although the quantification value of hGH by amino acid analysis showed good robustness in various conditions, results of intact protein analysis showed degradation profiles in temperatures higher than 4 °C. Consequently, storage and dissemination conditions should be set in accordance with stability tests. Based on the results, this method is believed to be suitable for accurate quantification of hGH. Additionally, it can also be used as a guide to preparation of CRM, and instructions for quality management of protein work for other similar proteins., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: True neurogenic thoracic outlet syndrome (TN-TOS) is an extremely rare neuromuscular disease. We report clinical, electrodiagnostic and radiologic features of patients with TN-TOS., Methods: Retrospective chart review of patients satisfying criteria was done. Nerve conduction study (NCS) and needle electromyography (EMG) of upper extremity were reviewed. Brachial plexus MRI and computed tomography angiography (CTA) were also reviewed., Results: Thirteen TN-TOS patients were identified. The most common neurologic signs were hypesthesia in the medial forearm or ulnar digits and weakness of the abductor pollicis brevis (APB) muscle. In NCS, medial antebrachial cutaneous (MABC) sensory nerve action potential amplitude was decreased in all tested patients. The APB muscle was most commonly involved in EMG. Among radiologic criteria, focal stenosis of subclavian artery in CTA was the most common finding., Conclusion: We confirmed that TN-TOS is T1 predominant lower roots/trunk brachial plexopathy with clinical and electrodiagnostic features. Radiologic studies may be used to detect structural abnormalities., Significance: As MABC NCS showed abnormal results in all tested patients, it should be added to electrodiagnostic study as screening method. If present, structural abnormalities might be confirmed with radiologic studies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: The tip of the tracheal tube should lie at the mid-tracheal level after tracheal intubation in paediatric patients. Auscultation does not guarantee optimal positioning of the tracheal tube. We compared auscultation and the ultrasound-guided lung sliding sign to confirm optimal positioning of the tracheal tube in paediatric patients., Methods: We studied 74 paediatric patients aged 0-24 months of ASA physical status 1-3 who were scheduled for elective surgery under general anaesthesia. All were randomly assigned to one of two groups: depth of tracheal tube confirmed by auscultation (Group A) or using the ultrasound-guided lung sliding sign (Group S)., Results: Optimal positioning of the tracheal tube was observed in 32 of 37 (87%) subjects in Group S and 24 of 37 (65%) subjects in Group A (difference in proportion, 22%; 95% confidence interval, 2-39%; P=0.030). Optimal depth correlated with patient height (adjusted coefficient=0.888, P<0.001)., Conclusions: In paediatric patients younger than 24 months, use of the ultrasound-guided lung sliding sign was more accurate than auscultation for optimal positioning of the tracheal tube., Clinical Trial Registration: KCT 0003015., (Copyright © 2019 British Journal of Anaesthesia. Published by Elsevier Ltd. All rights reserved.)

Bisphenol A (BPA), a synthetic monomer commonly included in the daily products, has a structure similar to the estrogen receptor agonist. Therefore BPA has been anticipated to interfere with the hormone metabolisms and cause diverse pathological conditions. But the effects of BPA on the genetic landscapes of liver or hepatic cells have not been fully established. Gene expressional changes induced by low- or high-dose of BPA were evaluated in 3D cultured human hepatoma cells (HepG2 spheroids) in vitro at 0, 0.5, 5 and 200 μM and liver of rats exposed to BPA at 0, 0.5 and 250 mg/kg for 90 days in vivo. Functional enrichment analysis, pathway activity measurement and network analysis were performed using BPA-responsive genes. Treatment with BPA changed a lot of gene expressions in both HepG2 spheroids and rat livers depending on doses of BPA. Functional enrichment and pathway analysis show that lipid or steroid metabolism-related functions were altered by BPA in both HepG2 spheroids and livers of rats. Lipid metabolism-related functions altered by BPA formed a large cluster encompassing lipid biosynthesis, steroid metabolic process and cholesterol regulation process. It was also observed that distribution of pathway activities was correlated between HepG2 spheroids and rat livers at low-dose of BPA. Distance distribution in protein-protein interaction network also evidenced the closeness of BPA-responsive genes to metabolism pathways which include lipid metabolism. Collectively, we demonstrated that BPA greatly influenced overall gene expression and biological functions in both human hepatoma spheroids and rat liver, in which lipid- or steroid metabolism-associated genes were significantly altered by the exposure to BPA., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: Risk factors of infection after deep brain stimulation (DBS) surgery in patients with Parkinson disease (PD) have been controversial. We aimed to investigate the clinical characteristics and risk factors of infection after DBS surgery in PD patients., Methods: We retrospectively investigated 246 consecutive DBS surgeries in 169 advanced PD patients. Clinical data were collected and analyzed to clarify the clinical characteristics associated with infection after DBS surgery. Multivariate logistic regression analysis was used to assess risk factors for infection after DBS surgery., Results: Infection occurred in 5% of all DBS surgeries and in 7% of all PD patients who received DBS surgery. Most infections (75%) occurred within 3months after DBS surgery but it also occurred 21months after DBS surgery. Gram-positive bacteria were the most common pathogens (75%). Infection after DBS surgery was associated with short period of prophylactic antibiotic therapy (OR=0.62, 95% CI=0.45-0.85, P=0.002) and intensive care unit (ICU) management immediate after DBS surgery (OR=5.43, 95% CI=1.12-26.45, P=0.036)., Conclusion: Our study suggests that short period of prophylactic antibiotic therapy and ICU management after surgery may increase the risk of infection in PD patients who received DBS surgery., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Astragali Radix, the root of Astragalus membranaceus Bunge, is one of the most frequently used crude drugs in Asian medicine. We developed a quantification method for 6 components (calycosin, formononetin, astragaloside I-IV) of Astragali Radix and Hwanggi-gyeji-omul-tang (HGOT) using reverse-phase high-performance liquid chromatography coupled with integrated pulsed amperometric detection (RP-HPLC-IPAD). The plants were extracted in 80% ethanol for 2h. All target components were detected with good sensitivity using sodium hydroxide (as a post-column eluent). The limit of detection (S/N=3) and limit of quantification (S/N=10) of the target components ranged from 0.10-1.00ng and from 0.30-3.00ng, respectively. The coefficients of linear regression ranged from 0.9993-1.0000, all interday and intraday precision values were <3.64%, and the average recovery ranged from 99.00-102.97% for Astragali Radix and 97.73-102.57% for HGOT. This method exhibited good selectivity, sensitivity, and reproducibility and can be used directly without any pretreatment steps. Our method will therefore be useful as a quality control measure for Astragali Radix., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Serotonin is known to be present in pancreatic β-cells and to play several physiological roles, including insulin secretion, β-cell proliferation, and paracrine inhibition of α-cells. However, the serotonin production of different cell lines and islets has not been compared based on age, sex, and diabetes related conditions. Here, we directly compared the serotonin concentrations in βTC and MIN6 cell lines, as well as in islets from mice using ultra-performance liquid chromatography tandem mass spectrometry. The average serotonin concentration was 5-10 ng/mg protein in the islets of male and non-pregnant female mice. The serotonin level was higher in females than males at 8 weeks, although there was no difference at 1 year. Furthermore, we observed serotonin by immunofluorescence staining in the pancreatic tissues of mice and human. Serotonin was detected by immunofluorescence staining in a portion of β-cells from islets of old female mice, but not of male or young female mice. A similar pattern was observed in human pancreas as well. In humans, serotonin production in β-cells was associated with a diabetes-free condition. Thus, serotonin production in β-cells was associated with old age, female sex, and diabetes-free condition., (Copyright © 2017 Elsevier Inc. All rights reserved.)

DA-9805 is a new botanical antiparkinson drug candidate formulated using an ethanolic extract of the root of Bupleurum falcatum, the root cortex of Paeonia suffruticosa, and the root of Angelica dahurica. In this study, a sensitive and rapid LC-MS/MS method was developed to simultaneously determine, saikosaponin a, paeonol, and imperatorin, three active/representative ingredients of DA-9805, in rat plasma. Plasma was extracted by mixture of ethyl acetate and methyl tertiary butyl ether. Chromatographic separation was carried out using a C 18 column and a gradient elution of mobile phases consisting of 5mM formic acid in water and acetonitrile. Total chromatographic run time was 10.5min. Multiple reaction monitoring mode was used for mass spectrometry; the transitions were m/z 779.5→617.2 for saikosaponin a in negative-ion mode, m/z 167→149 for paeonol and m/z 271.1→203 for imperatorin in positive-ion mode. Calibration curves were constructed in the range of 0.5-1000ng/mL for saikosaponin a, 20-10000ng/mL for paeonol, and 0.2-1000ng/mL for imperatorin. All the validation data, including the selectivity, linearity, precision, accuracy, recovery, matrix effect, and stability satisfied the acceptance requirements. The method was successfully applied in a pharmacokinetic study of saikosaponin a, paeonol, and imperatorin following oral administration of DA-9805., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Glycated hemoglobin (HbA 1c ), defined as hemoglobin (Hb) molecules having a stable adduct of glucose on the N-terminal of the β-chains, has been endorsed as a diagnostic tool for diabetes mellitus and a prediction indicator for the development of diabetes complications. Here we describe an accurate procedure using two stages of isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) for the quantification of HbA 1c that provides full traceability to International System of Units (SI). First, synthetic peptides representing specific markers of HbA 1c (G-hexa) and hemoglobin A 0 (Hexa) were certified by amino acid analysis via acid hydrolysis as reference materials (RMs) for the next step. For this peptide certification, three amino acids (proline, valine, and leucine) were determined by hydrolysis with 10M hydrochloric acid at 130°C for 48h followed by ID-LC-MS/MS. Then, HbA 1c content in blood was quantified with the ratio of specific proteolytic peptides from HbA 1c and HbA 0 via enzyme digestion using ID-LC-MS/MS with the certified peptides as RMs and isotope-labeled peptides as internal standards. Results demonstrate complete traceability to SI-units throughout this procedure. Reliability was confirmed through comparative studies with commercially available RMs for HbA 1c , and other routine HbA 1c diagnostic methods as well. Following full method validation, we applied this procedure to the certification of candidate hemolysate-certified RMs for HbA 1c content, as well as 52 real clinical samples. All of the results showed the suitability of this method to act as a primary reference measurement procedure for HbA 1c in complex biological samples., (Copyright © 2017 Elsevier B.V. All rights reserved.)

We described a reference measurement procedure for amino acid (AA) quantification in blood samples based on deproteinization with 5-sulfosalicylic acid (SSA) and an isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (LC-MS) method. The serum was deproteinized with 15% v/v SSA and the supernatant was injected directly into the LC-MS system without further processing. Compared with the use of other precipitants and water as a control, five model AAs-valine, isoleucine, leucine, tyrosine, and phenylalanine-in the SSA-treated samples showed ionization enhancement as well as stable background signals without significant ion suppression effects. Five analytes were clearly separated within 3min using gradient elution and ion-pair chromatography of water and acetonitrile containing 0.1% v/v trifluoroacetic acid. The limit of detection range of this method was 2-52fmol, and the RSDs of accuracy and precision from intra- and inter-day assays were within 2.7%. The method was applied to various blood samples including serum, whole blood and plasma, with no reasonable measurement bias revealed. The quantification accuracy of this method was then assessed using commercially available plasma certified reference material (CRM) for AA, and the results agreed well within certified values. We finally applied this method to the determination of candidate serum CRM. The optimized protocol was found to be suitable for the accurate quantification of five AAs in serum, and may satisfactorily serve as a primary method for AA measurement in various blood matrices., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The ban on the use of antibiotics as growth promoters due to resistance issues has urged scientists to find alternatives to antibiotics. Entercoccus faecium is one of the probiotics which have been used as an alternative to antibiotics in the livestock industry. This study was conducted to evaluate the effect of probiotic (Enterococcus faecium DSM 7134) supplementation on production performance, feed intake, egg quality, excreta microflora, ammonia emission, and nutrient utilization in laying hens. A total of 288 ISA brown laying hens were used in a 27 wk feeding experiment and randomly assigned to 3 dietary treatments with 8 replicates of 12 birds each. The treatments were CON (basal diet), PB1 (basal diet + 0.005% E. faecium), and PB2 (basal diet + 0.01% E. faecium). Overall, our results demonstrated that E. faecium supplementation resulted in a significant increase in egg production, egg shell thickness, and nutrient digestibility (dry matter, nitrogen, and energy) in laying hens, and a significant reduction in fecal coliform counts as compared with CON. The shift of excreta fecal microbial composition by E. faecium supplementation was accompanied by increased nutrient retention and reduction in nutrient excretion, leading to improved nutrient digestibility and reduced excreta ammonia emission. Overall, E. faecium supplementation appears to have a beneficial effect in ISA brown laying hens and should be considered as a positive diet supplement to use in the industry., (© 2016 Poultry Science Association Inc.)

Aims: This study's purpose was to evaluate the effect of ultrasound in air at a frequency of 0.04MHz and an intensity of 50mW/cm 2 on neural differentiation of hBM-MSCs in vitro and on neurogenesis in an ischemic stroke model., Materials and Methods: hBM-MSCs were exposed to 0.04MHz ultrasound and then compared with no exposed one in cell morphology, lactate dehydrogenase (LDH) activity, RT-PCR, and Western blot. In addition, we made stroke model mice by means of the photothrombosis (PT) method and these models were exposed to 0.04MHz ultrasound after hBM-MSCs injection. We compared with sham group in histological and immunohistochemical analysis and western blot., Key Findings: Ultrasound induced neural differentiation without cell death. In stroke models, inflammatory cells were observed around the infarct area in the Cell, Cell/Ultrasound group and the brain infarct volume in the Cell/Ultrasound group was smaller than in the sham group. Further, the expression of neural proteins in the Cell/Ultrasound group was increased relative to the sham group., Significance: The present study showed that ultrasound promotes neural differentiation of hBM-MSC and neurogenesis in a mouse stroke model. This may be applicable as a therapeutic device with the aim of inducing neurogenesis following stroke., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Isotopically labeled proteins have been used as internal standards for mass spectrometry (MS)-based absolute protein quantification. Although this approach can provide highly accurate analyses of proteins of interest within a complex mixture, one of the major limitations of this method is the difficulty in preparing uniformly labeled standards. Human growth hormone (hGH) is one of the most important hormones that circulate throughout the body, and its measurement is primarily of interest in the diagnosis and treatment of growth disorders. In order to provide a useful internal standard for MS-based hGH measurement, we describe an efficient strategy to produce a potentially valuable, stable isotope-labeled hGH with high purity and yield. The strategy involves the following steps: solubilization of hGH under labeling conditions, detection of stable isotope incorporation, large-scale purification, analysis of the labeled protein, and assessment of the labeling efficiency. We show that the yield of soluble hGH under selective isotopic labeling conditions can be greatly increased by optimizing protein expression and extraction. Our efficient method for generating isotopically labeled hGH does not influence the structural integrity of hGH. Finally, we assessed the efficiency of stable isotope labeling at the intact protein level, and the result was further verified by amino acid analysis. These results clearly indicate that our labeling approach allows an almost complete incorporation of 13 C 6 15 N 4 -arginine into the hGH expressed in E.coli without detectable isotope scrambling., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Pemphigus represents a group of organ specific autoimmune blistering disorders of the skin mediated by pathogenic autoantibodies with well-defined antigenic targets. While most of these diseases are sporadic, endemic forms of disease do exist. The endemic form of pemphigus foliaceus (also known as fogo selvagem, FS) exhibits epidemiological features that suggest exposure to hematophagous insect bites are a possible precipitating factor of this autoimmune disease, and provides a unique opportunity to study how environmental factors contribute to autoimmune disease development. FS patients and healthy individuals from endemic regions show an autoreactive IgM response that starts in early childhood and becomes restricted to IgG4 autoantibodies in FS patients. In searching for triggering environmental antigens, we have found that IgG4 and IgE autoantibodies from FS patients cross-react with a salivary antigen from sand flies. The presence of these cross-reactive antibodies and antibody genetic analysis confirming that these antibodies evolve from the same naïve B cells provides compelling evidence that this non-infectious environmental antigen could be the initial target of the autoantibody response in FS. Consequently, FS serves as an ideal model to study the impact of environmental antigens in the development of autoimmune disease., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The increased volumes of carbon nanotubes (CNTs) being utilized in industrial and biomedical processes carries with it an increased risk of unintentional release into the environment, requiring a thorough hazard and risk assessment. In this study, the toxicity of pristine and hydroxylated (OH-) multiwall CNTs (MWCNTs) was investigated in the nematode Caenorhabditis elegans using an integrated systems toxicology approach. To gain an insight into the toxic mechanism of MWCNTs, microarray and proteomics were conducted for C. elegans followed by pathway analyses. The results of pathway analyses suggested endocytosis, phagocytosis, oxidative stress and endoplasmic reticulum (ER) stress, as potential mechanisms of uptake and toxicity, which were subsequently investigated using loss-of-function mutants of genes of those pathways. The expression of phagocytosis related genes (i.e. ced-10 and rab-7) were significantly increased upon exposure to OH-MWCNT, concomitantly with the rescued toxicity by loss-of-function mutants of those genes, such as ced-10(n3246) and rab-7(ok511). An increased sensitivity of the hsp-4(gk514) mutant by OH-MWCNT, along with a decreased expression of hsp-4 at both gene and protein level suggests that MWCNTs may affect ER stress response in C. elegans. Collectively, the results implied phagocytosis to be a potential mechanism of uptake of MWCNTs, and ER and oxidative stress as potential mechanisms of toxicity. The integrated systems toxicology approach applied in this study provided a comprehensive insight into the toxic mechanism of MWCNTs in C. elegans, which may eventually be used to develop an "Adverse Outcome Pathway (AOP)", a recently introduced concept as a conceptual framework to link molecular level responses to higher level effects., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Background & Aims: Most studies on the role of STAMP2 in metabolism have used adipose tissue. Little knowledge exists concerning the role of STAMP2 in the liver, which is a metabolically central target. We hypothesized that STAMP2 is involved in non-alcoholic fatty liver disease (NAFLD) pathogenesis., Methods: We examined our hypothesis using human NAFLD patient pathology samples and a high-fat diet (HFD)-induced NAFLD mouse model. The molecular mechanism underlying hepatic STAMP2-mediated lipid imbalance was explored using an oleic acid (OA)-induced NAFLD in vitro model., Results: Noticeably, the expression level of STAMP2 protein was reduced in the livers obtained from NAFLD patients and HFD-induced NAFLD mice. In vivo knockdown of hepatic STAMP2 by siRNA accelerated hepatic steatosis and insulin resistance in mice fed a HFD. Conversely, the delivery of adenoviral STAMP2 (Ad-STAMP2) improved hepatic steatosis in HFD-induced NAFLD mice. The expression of lipogenic or adipogenic factors was increased in both in vitro and in vivo NAFLD models but was reversed by Ad-STAMP2. Adenoviral overexpression of STAMP2 improved insulin resistance in the HFD-induced NAFLD mice. In vivo and in vitro assays demonstrated that STAMP2 modulates insulin sensitivity and glucose metabolism and that STAMP2 counteracts OA-induced insulin resistance by modulating insulin receptor substrate-1 stability., Conclusions: The present study revealed that hepatic STAMP2 plays a pivotal role in preventing HFD-induced NAFLD and that STAMP2 overexpression improves hepatic steatosis and insulin resistance in NAFLD. Our findings indicate that STAMP2 may represent a suitable target for interventions targeting NAFLD., (Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

Bacillus subtilis C-3102 has been used as a direct-fed microbial or probiotic product since 1986 to improve production performance in broilers worldwide. This study was conducted to determine and confirm the effect of B. subtilis C-3102 spore supplementation to feed on growth performance, nutrient digestibility, carcass quality, blood profiles, noxious gas emission, and intestinal and excreta microflora in broilers. A total of 816 one-day-old male Ross 308 broilers (46.06 ± 0.67 g) were used in a 5-wk study with Calsporin, B. subtilis final product (1.0 × 10(9) cfu/g of B. subtilis). Broilers were randomly allotted to 1 of 3 dietary treatments consisting of 16 replicate cages with 17 broilers each: I) CON (control, basal diet), II) BS300 (CON + 300 mg of B. subtilis/kg of feed), and III) BS600 (CON + 600 mg of B. subtilis/kg of feed). Regarding probiotic effect, B. subtilis significantly increased Lactobacillus counts in the cecum, ileal, and excreta, and reduced Escherichia coli counts in the cecum and excreta, compared with CON. In addition, supplementation also tended to reduce Clostridium perfringens counts in the large intestine and excreta, while linearly reducing Salmonella counts in the cecum, ileal, large intestine, and excreta, compared with CON. Regarding growth performance, B. subtilis enhanced ADG in the starter and overall experimental periods, without any effects on feed intake compared with CON. Consequently, feed conversion ratio in the grower-finisher and overall experimental periods decreased significantly. The inclusion of B. subtilis improved the digestibility of DM and gross energy, as well as reducing ammonia emission, compared with CON. No significant difference in breast muscle color, water holding capacity, and drip loss, and relative organ weights, as well as in white blood cells, red blood cells, lymphocyte counts, and IgG amount, were observed. Overall, B. subtilis C-1302 is capable of providing a probiotic effect leading to improved growth performance and feed efficiency, due to the manipulation of intestinal microflora, with minimal side effects in broilers., (©2014 Poultry Science Association Inc.)

A prospective alternative to antibiotics currently being evaluated is yeast and its derivative products. Phaffia rhodozyma is a species of yeast that produces the carotenoid pigment, astaxanthin (AST), which exhibits a wide variety of biological activities, including antioxidation in animals. A total of 432 one-day-old male broilers (Arbor Acres) were used in a 4-wk feeding experiment and each dietary treatment consisted of 9 replicate cages, with 16 broilers per replicate. Birds were randomly allotted to 1 of 3 corn-soybean meal-based diets supplemented with 0 mg (CON, basal diet), 1,000 mg (CON + AST production 0.1%), or 2,000 mg (CON + AST production 0.2%) of P. rhodozyma yeast per kg of feed, giving an intake of approximately 0, 2.3, and 4.6 mg of AST/kg of feed, respectively. The inclusion of AST linearly improved weight gain in the finisher period (linear, P = 0.0264) and during the overall experimental period (linear, P = 0.0194) and linearly decreased feed conversion ratio in the finisher period (linear, P = 0.0422) and tended to decrease during the overall experimental period (linear, P = 0.0568). No significant effects were observed with red blood cell, white blood cell, and lymphocyte numbers in response to 2.3 or 4.6 mg of AST/kg of feed (P > 0.05). The ammonia emission from samples treated with 2.3 and 4.6 mg of AST/kg was significantly lower than that of CON (linear, P = 0.0110). Taken together, these results indicate that supplementation with AST could improve BW gain and decrease feed conversion ratio and fecal noxious gas emission of ammonia in broilers., (©2014 Poultry Science Association Inc.)

Objective: To evaluate the change in mandibular second-molar (M2) angulation in orthodontic treatment with premolar extraction and lower third molar (M3)., Study Design: Panoramic radiographs were evaluated in 3 groups of 129 participants: (1) control, no orthodontic treatment (n = 65); (2) extraction, orthodontic treatment with premolar extraction (n = 30); and (3) nonextraction, orthodontic treatment without premolar extraction (n = 34). The angular difference and ratio of M2 to the first molar (M1), the change in the angulation of M2 between pre- and postorthodontic treatment, and the distal bone level of M2 were assessed., Results: The angular difference and ratio of M2 to M1, the angulation change of M2 between pre- and postorthodontic treatment, and the distal bone level of M2 were higher in the nonextraction group than in the control and extraction groups., Conclusions: The successful orthodontic alignment of the M2 may not be achieved in nonextraction cases when the M3s are present., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Neutrophil gelatinase-associated lipocalin (NGAL) exists as monomers, homodimers, and NGAL/matrix metalloproteinase-9 (MMP-9) complexes. Circulating neutrophils are activated in ongoing disseminated intravascular coagulation (DIC); therefore, plasma NGAL levels are likely to be increased in DIC. We investigated the diagnostic performance of the plasma NGAL level in predicting acute kidney injury (AKI) in patients with DIC and determined the prognostic value of NGAL., Methods: A total of 126 patients with suspected DIC were enrolled. The plasma free NGAL was analyzed with a point-of-care immunoassay. Plasma total NGAL and NGAL/MMP-9 complex levels were measured using commercial kits., Results: Median free and total NGAL levels were markedly higher in patients with AKI than those without AKI. However, no significant difference in the NGAL/MMP-9 complex level was found between the groups. In patients without AKI, the plasma free and total NGAL levels were significantly higher in those with overt-DIC than in those without overt-DIC. Of note, the free NGAL level showed a significant prognostic value in DIC., Conclusion: Plasma free and total NGAL proved to be powerful markers for AKI in DIC patients and plasma free NGAL also has a prognostic relevance in DIC patients without AKI., (Copyright © 2014. Published by Elsevier B.V.)

Management of patella infera remains a difficult therapeutic endeavor. We report a case of a 21-year-old man, who had development of patella infera and knee flexion contracture after a patella fracture. Patella infera was treated by patellar tendon lengthening and augmentation with a tibialis anterior tendon allograft fixed with bioabsorbable cross-pins. The patient regained an anatomic patellar position and full range of motion at two years after surgery., (© 2013.)

The formation of amyloid β (Aβ) fibrils is crucial in initiating the cascade of pathological events that culminates in Alzheimer's disease. In this study, we investigated the mechanism of Aβ fibril formation from hydrodynamically well defined species under controlled aggregation conditions. We present a detailed mechanistic model that furnishes a novel insight into the process of Aβ42 fibril formation and the molecular basis for the different structural transitions in the amyloid pathway. Our data reveal the structure and polymorphism of Aβ fibrils to be critically influenced by the oligomeric state of the starting materials, the ratio of monomeric-to-aggregated forms of Aβ42 (oligomers and protofibrils), and the occurrence of secondary nucleation. We demonstrate that monomeric Aβ42 plays an important role in mediating structural transitions in the amyloid pathway, and for the first time, we provide evidences that Aβ42 fibrillization occurs via a combined mechanism of nucleated polymerization and secondary nucleation. These findings will have significant implications to our understanding of the molecular basis of amyloid formation in vivo, of the heterogeneity of Aβ pathology (e.g., diffuse versus amyloid plaques), and of the structural basis of Aβ toxicity., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The Bradford assay is a simple method for protein quantitation, but variation in the results between proteins is a matter of concern. In this study, we compared and normalized quantitative values from two models for protein quantitation, where the residues in the protein that bind to anionic Coomassie Brilliant Blue G-250 comprise either Arg and Lys (Method 1, M1) or Arg, Lys, and His (Method 2, M2). Use of the M2 model yielded much more consistent quantitation values compared with use of the M1 model, which exhibited marked overestimations against protein standards., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Purpose: This was a cross-sectional study designed to evaluate the subjective understanding of technical terms and contents of the informed consent forms given to patients about to undergo conscious sedative upper gastrointestinal endoscopy., Methods: A group of conveniently selected 180 patients, who were undergoing sedative endoscopy were recruited in the endoscopy procedure room in a tertiary hospital from June to July 2011. Data were collected with a structured questionnaire., Results: The average number of terms exposed was 8.12 out of 10 items, the average number of terms understood well by the patients was 5.53 out of 10 items, and the average number of right answers was 3.30 out of 5 items, and the percent of correct answers ranged from 26.1% to 90.0%. The exposure to terms differed by gender, education, and previous exposure to sedative endoscopy procedures. The number of "understanding of the terms well" responses differed according to age and previous exposure to sedative endoscopy procedures, and the correct answer rate was differed by education., Conclusion: The understanding of the terms and knowledge about the procedures were disappointing. Therefore, sufficient explanations should be provided to the patients. While the informed consent was taken by doctors, the level of understanding should be monitored by nurses. In particular, subjects who did not have any previous experience with endoscopy procedures showed relatively lower level of understanding. We recommend that medical terms should be replaced with more common and nontechnical words in consent forms., (Copyright © 2013. Published by Elsevier B.V.)

In an effort to identify novel genes related to the prognosis of gastric cancer, we performed gene expression profiling and found overexpressed levels of human interferon-induced transmembrane protein 1 (IFITM1). We validated the gastric cancer-specific up-regulation of IFITM1 and its association with cancer progression. We also studied its epigenetic regulation and tumorigenesis-related functions. Expression of IFITM1 was evaluated in various human gastric cancer cells and in 35 patient tumor tissues by quantitative RT-PCR and Western blot analyses. The results showed highly up-regulated IFITM1 in cancer cell lines and tissues. Furthermore, IHC studies were performed on 151 patient tissues, and a significant correlation was revealed between higher IFITM1 expression and Lauren's intestinal type (P = 0.007) and differentiated adenocarcinoma (P = 0.025). Quantitative studies of DNA methylation for 27 CpG sites in the regulatory region showed hypermethylation in cells expressing low levels of IFITM1. Methylation-dependent IFITM1 expression was confirmed further by in vitro demethylation using 5-aza-2'-deoxycytidine and luciferase assays. The functional analysis of IFITM1 by silencing of its expression with small-interfering RNA showed decreased migration and invasiveness of cancer cells, whereas its overexpression exhibited the opposite results. In this study, we demonstrated gastric cancer-specific overexpression of IFITM1 regulated by promoter methylation and the role of IFITM1 in cancer prognosis., (Copyright © 2012 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

Objectives: The objective of this histologic study was to evaluate platelet-rich fibrin (PRF)-mixed tricalcium phosphate (TCP) and recombinant human bone morphogenic protein 2 (rhBMP-2)-coated TCP in their potential to enhance bone regeneration in sinus elevation in rabbits as well as in their inflammatory features., Study Design: Bilateral round-shaped defects (diameter 8.0 mm) were formed in the maxillary anterior sinus walls of 36 New Zealand white rabbits. The defects were grafted with TCP only (control group), with rhBMP-2-coated TCP (experimental group A) and with PRF-mixed TCP (experimental group B). Each group included 12 rabbits. The animals were killed at 3 days, 1 week, 2 weeks, 4 weeks, 6 weeks, and 8 weeks. The specimens underwent decalcification and were stained for histologic analysis., Results: There were no significant differences in inflammatory features among the groups at 3 days or the first week after operation. In a histomorphometric analysis, the new bone formation ratio showed significant differentiation between groups A and B. The TCP-only control group showed a relatively lower bone formation ratio rather than the experimental groups. The PRF-mixed TCP group showed a larger bone formation area, compared with both the control group and group A., Conclusions: In the results of the histologic evaluation (hematoxylin-eosin, Masson trichrome stain), the experimental groups A and B showed rapid bone formation, remodeling, and calcification in the second week. Moreover, there was a significant difference between those experimental groups and the control group in the new bone formation area at the fourth, sixth, and eighth weeks. The PRF-mixed TCP showed more rapid bone healing than the rhBMP-2-coated TCP or the TCP-only control., (Copyright © 2012 Elsevier Inc. All rights reserved.)