•A new bioanalytical HPLC-FL method to determine velpatasvir (VEL).•The method offered sufficient sensitivity, comparable to LC-MS/MS methods.•VEL exhibited moderate intestinal permeability and ...significant biliary excretion.•VEL was significantly metabolized in the liver, but not in the intestine.•Gut absorption and hepatic first-pass metabolism mainly determined the oral bioavailability of VEL.
Velpatasvir is a novel inhibitor of hepatitis C virus nonstructural protein 5A that received US Food and Drug Administration approval for the treatment of patients with chronic hepatitis C virus genotypes 1–6. In the present study, a sensitive bioanalytical method for velpatasvir was developed using high-performance liquid chromatography coupled with a fluorescence detector system, which was applied to elucidate the factors determining the oral bioavailability and disposition of velpatasvir. This method offered sufficient sensitivity, with a lower limit of quantification of 0.5 ng/mL, which is comparable to previously reported methods using liquid chromatography coupled with tandem mass spectrometry. Velpatasvir exhibited low oral bioavailability, moderate intestinal permeability, and significant biliary excretion in rats. It was also found to be significantly metabolized in the liver, with a low-to-moderate extraction ratio; however, its intestinal metabolism and enterohepatic circulation did not occur. Thus, our present results demonstrate that the oral bioavailability of velpatasvir is primarily dependent on gut absorption and hepatic first-pass metabolism. The fractions of velpatasvir dose unabsorbed from the gut and eliminated by the liver before reaching the systemic circulation following oral administration were estimated to be 32.8%–58.6% and 4.74%–30.54% of the oral dose, respectively. To our knowledge, this is the first systematic study to investigate the contributory roles of biopharmaceutical and pharmacokinetic factors on the oral bioavailability of velpatasvir, together with a new bioanalytical method for velpatasvir.
In this study, a novel polyhistidine‐incorporated lipid nanoparticle (pHis/LNP) is developed for the delivery of therapeutic globotriaosylceramide (Gb3) synthase siRNAs using a microfluidic device ...with pHis as a biocompatible method of endosome escape. To inhibit the expression of Gb3 synthase, six siRNAs against Gb3 synthase are designed and an optimal siRNA sequence is selected. Selected Gb3 synthase siRNA is incorporated into pHis/LNP to prepare a spherical siRNA pHis/LNP with a size of 62.5 ± 1.9 nm and surface charge of −13.3 ± 4.2 mV. The pHis/LNP successfully protects siRNAs from degradation in 50% serum condition for 72 h. Prepared pHis/LNP exhibits superior stability for 20 days and excellent biocompatibility for A549 cells. After treatment with fluorescence‐labeled LNPs, dotted fluorescent signals are co‐localized with Lysotracker in cells with LNPs, whereas strong and diffused fluorescence intensity is observed in cells with pHis/LNPs probably due to successful endosomal escape. The extent of Gb3 synthase gene silencing by siRNA pHis/LNP is greatly improved (6.0‐fold) compared to that by siRNA/LNP. Taken together, considering that the fabricated siRNA pHis/LNP exhibits excellent biocompatibility and superior gene silencing activity over conventional LNP, these particles can be utilized for the delivery of a wide range of therapeutic siRNAs.
In this study, a novel polyhistidine‐incorporated lipid nanoparticle (pHis/LNP) for the delivery of siRNAs is successfully developed. The fabricated siRNA pHis/LNPs exhibited excellent biocompatibility and superior gene silencing activity due to high rate of endosome escape by polyhistidine, which can be utilized for the delivery of a wide range of therapeutic siRNAs.
Background Globally, ischemic stroke is a major health threat to humans that causes lifelong disability and death. Mentha arvensis (MA) has been used in traditional medicine to alleviate oxidative ...stress and inflammation-related disorders. In the present study, the neuroprotective properties of fermented MA (FMA) extract were investigated in the gerbil and SH-SY5Y cells. model of transient global cerebral ischemia. Methods Bilateral common carotid artery occlusion-induced transient global cerebral ischemia in gerbil and hydrogen peroxide (H.sub.2O.sub.2)-mediated neurotoxic effects in human neuroblastoma cells (SH-SY5Y) were investigated. FMA (400 mg/kg) was orally administered for 7 days before induction of ischemic stroke. To evaluate the neuroprotective activity of FMA, we implemented various assays such as cell viability assay (MTT), lactate dehydrogenase (LDH) assay, histopathology, immunohistochemistry (IHC), histofluorescence, and western blot. Results FMA pretreatment effectively decreased transient ischemia (TI) induced neuronal cell death as well as activation of microglia and astrocytes in the hippocampal region. The protective effects of FMA extract against H.sub.2O.sub.2-induced cytotoxicity of SH-SY5Y cells were observed by MTT and LDH assay. However, FMA pretreatment significantly increased the expression of the antioxidant marker proteins such as superoxide dismutase-1 (SOD-1) and superoxide dismutase-2 (SOD-2) in the hippocampus and SH-SY5Y cells. Furthermore, the activation of mitogen-activated protein kinase (MAPK) further activated a cascade of outcomes such as neuroinflammation and apoptosis. FMA pretreatment notably decreased TI and H.sub.2O.sub.2 induced activation of MAPK (c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), and p38) proteins in hippocampus and SH-SY5Y cells respectively. Besides, pretreatment with FMA markedly reduced H.sub.2O.sub.2 mediated Bax/Bcl2 expression in SH-SY5Y cells. Conclusion Thus, these results demonstrated that neuroprotective activities of FMA might contribute to regulating the MAPK signaling pathway. Keywords: Fermented Mentha arvensis (FMA), Ischemic stroke, Neuroprotection, Antioxidant, MAPK
This study investigated the role of BRAF mutation analysis in thyroid fine-needle aspiration (FNA) samples compared to ultrasonographic and cytological diagnoses. A total 316 patients underwent ...ultrasonography (US)-guided FNA with BRAFV600E mutation analysis to diagnose thyroid nodules. One hundred sixteen patients with insufficient US images (n = 6), follow-up loss (n = 43), or unknown final diagnosis (n = 67) were excluded from the study. Comparisons between US diagnoses, cytological diagnoses, and BRAF mutation analysis were performed. Of 200 thyroid nodules, there was US diagnosis with 1 false negative and 11 false positive cases, cytological diagnosis with 10 false negative and 2 false positive cases, and BRAFV600E mutation analysis with 19 false negative and 2 false positive cases. The sensitivity, specificity, positive and negative predictive values, and accuracy of BRAFV600E mutation analysis were 83.2%, 98.1%, 97.5%, 86.6%, and 91%, respectively. Of the 18 nodules with Bethesda category III, 9 were true positive, 6 were true negative, 3 was a false negative, and none were false positive on BRAF mutation analysis. In conclusion, we recommend that BRAFV600E mutation analysis only be performed for evaluating thyroid nodules with Bethesda category III, regardless of US diagnosis.
Anti-cyclic citrullinated peptide antibody testing is used to diagnose rheumatoid arthritis and associated with interstitial lung disease in RA. Herein, we investigate the relationship between ...anti-CCP antibody and ILD in SSc. We performed a retrospective analysis at a tertiary medical center between 2005 and 2019. Patients with SSc, systemic lupus erythematosus, and polymyositis/dermatomyositis (PM/DM) were evaluated for anti-CCP antibody and ILD. Additionally, medical records of SSc patients with ILD were reviewed. SSc patients had the highest anti-CCP antibody positivity rate compared to those with SLE and PM/DM. The incidence of ILD was higher in SSc patients with anti-CCP antibody than in those without. The usual interstitial pneumonia (UIP) incidence was higher in the anti-CCP antibody-positive group than in the anti-CCP antibody-negative group. The DLCO was lower in the anti-CCP antibody-positive group than in the anti-CCP antibody-negative group. On multivariable analysis, factors associated with SSc-ILD were anti-CCP antibody or rheumatoid factor (β coefficient, 2.652 95% CI 1.472 to 4.776) and anti-Scl70 antibody (β coefficient, 4.011 95% CI 2.142 to 7.508). Anti-CCP antibody may be associated with a higher incidence of ILD in SSc. SSc patients with anti-CCP antibody may have more UIP pattern and lower DLCO.Trial Registration Retrospectively registered.
Abstract Attention deficit hyperactivity disorder (ADHD) is caused by the interaction of genetic and environmental factors. The objective of this study was to examine the effects of prenatal exposure ...to alcohol and environmental tobacco smoke (ETS). Among the 30,552 parents who responded to a survey, the answers of 19,940 who replied to questions on prenatal exposure to ETS, alcohol consumption, and completed the DuPaul Rating Scale were analyzed. Results revealed that risk of ADHD significantly increased as a result of exposure to alcohol by 1.55 times (95% CI 1.33–1.82), maternal smoking during pregnancy by 2.64 times (95% CI 1.45–4.80), and paternal smoking during pregnancy by 1.17 times (95% CI 1.98–1.39). When the subjects whose mothers did not smoke during pregnancy were divided into 4 groups, the prevalence was 1.16 times higher (95% CI 1.02–1.33) in the group exposed to ETS but not alcohol, 1.19 times higher (95% CI 0.91–1.57) in the group exposed to alcohol but not ETS, and 1.58 times higher (95% CI 1.31–1.91) in the group exposed to ETS and alcohol. The differences between the groups were statistically significantly ( P <0.0001). This result shows that simultaneous exposure to ETS and alcohol during pregnancy increases the risk of ADHD.
The high-temperature mechanical behavior of cross-weld specimens prepared from a dissimilar weld joint between T92 martensitic and Super304H austenitic heat-resistant steels incorporating Ni-based ...weld metal was evaluated at temperatures up to 650°C. For both high temperature tensile and creep tests, failure took place in T92 due to its faster degradation with temperature increase. The heat-affected zone of T92 played a critical role during creep deformation, resulting in type IV failure under the long-term creep condition. For the creep specimens, the location of failure shifted from the base metal region to the fine-grained heat-affected zone as the creep duration time increased from the short-term to the long-term condition. The massive precipitation of Laves phase on the grain boundaries of the fine-grained heat-affected zone during creep deformation was observed and found to be responsible for the accelerated void formation in the area leading to the premature failure.
•Tensile and creep properties of the Super304H/T92 weld joint were evaluated.•The type IV brittle failure during long-term creep was carefully analyzed.•Near brittle cracks, massive formation of Laves phase was visualized by STEM-HAADF.•Also, these Laves phases were found to exist inside the creep-voids by EPMA.•This is the first microscopic evidence connecting Laves phase and creep-voids.
The hydrogen storage behavior and the microstructural features of AB-type Ti50Fe48V2 hydrogen storage alloys containing a small amount of cerium (Ce) were investigated to understand the effect of Ce ...addition during initial hydrogen absorption. The initial hydrogen absorption kinetics of the alloys improved significantly at room temperature with Ce addition, which exhibited no significant influence on the pressure-composition isotherms for hydrogen absorption and desorption. Fine spherical particles containing Ce, which were determined to be γ-Ce mixed with cerium oxide, were dispersed in the ordered body-centered cubic TiFe matrix. During the early stage of hydrogen absorption, small cracks were initiated around the Ce particles, likely caused by the volume expansion owing to the formation of ε-CeH2. Subsequently, many large cracks, believed to have formed owing to the hydrogenation of the TiFe matrix, propagated during further hydrogen absorption. Therefore, these Ce particles appear to play a crucial role by providing starting points for the initial hydrogenation, with this mechanism explaining the significant increase in the primary hydrogen absorption kinetics after Ce addition. Notably, some small pits were observed after partial hydrogen absorption, possibly attributed to the hydrogenation of Ce particles underneath the alloy surfaces.
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•Initial hydrogen absorption kinetics of TiFe alloys improved by Ce addition.•Ce addition forms Ce particles in alloys, playing a vital role during absorption.•Particle cracks formed owing to the formation of CeH2 during initial absorption.•Cracks propagate in the TiFe matrix during further absorption.•Small pits form during absorption owing to presence of CeH2 below alloy surface.
encodes an evolutionarily conserved widely expressed novel 8-pass transmembrane protein of unknown function in human. Here we identify
homozygous hypomorphic missense variants in our recessive ...polymicrogyria (PMG) cohort. Patients carrying
mutations exhibit striking neocortical PMG and intellectual disability.
knockout mice fail to develop midline hemispheric cleavage, whereas knock-in of patient mutations and patient-derived brain organoids show defects in apical cell polarity and radial glial scaffolding. We found that TMEM161B modulates actin filopodia, functioning upstream of the Rho-GTPase CDC42. Our data link
with human PMG, likely regulating radial glia apical polarity during neocortical development.
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