Direct Reprogramming of Rat Neural Precursor Cells and Fibroblasts into Pluripotent Stem Cells

Background: Given the usefulness of rats as an experimental system, an efficient method for generating rat induced pluripotent stem (iPS) cells would provide researchers with a powerful tool for studying human physiology and disease. Here, we report direct reprogramming of rat neural precursor (NP) cells and rat embryonic fibroblasts (REF) into iPS cells by retroviral transduction using either three (Oct3/4, Sox2, and Klf4), four (Oct3/4, Sox2, Klf4, and c-Myc), or five (Oct3/4, Sox2, Klf4, c-Myc, and Nanog) genes. Methodology and Principal Findings: iPS cells were generated from both NP and REF using only three (Oct3/4, Sox2, and Klf4) genes without c-Myc. Two factors were found to be critical for efficient derivation and maintenance of rat iPS cells: the use of rat instead of mouse feeders, and the use of small molecules specifically inhibiting mitogen-activated protein kinase and glycogen synthase kinase 3 pathways. In contrast, introduction of embryonic stem cell (ESC) extracts induced partial reprogramming, but failed to generate iPS cells. However, when combined with retroviral transduction, this method generated iPS cells with significantly higher efficiency. Morphology, gene expression, and epigenetic status confirmed that these rat iPS cells exhibited ESC-like properties, including the ability to differentiate into all three germ layers both in vitro and in teratomas. In particular, we found that these rat iPS cells could differentiate to midbrain-like dopamine neurons with a high efficiency. Conclusions/Significance: Given the usefulness of rats as an experimental system, our optimized method would be useful for generating rat iPS cells from diverse tissues and provide researchers with a powerful tool for studying human physiology and disease

A Feasibility Study for a Nonlinear Guided Wave Mixing Technique

Ultrasonic non-destructive testing is an effective means of examining objects without destroying them. Among such testing, ultrasonic nonlinear evaluation is used to detect micro-damage, such as corrosion or plastic deformation. In terms of micro-damage evaluation, the data that comes from amplitude comparison in the frequency domain plays a significant role. Its technique and parameter are called ultrasonic nonlinear technique and nonlinearity. A certain portion of nonlinearity comes from the equipment system, while the other portion of nonlinearity comes from the material. The former is system nonlinearity, while the latter is material nonlinearity. System nonlinearity interferes with interpretation, because its source is not from the material. In this study, in order to minimize system effects, a mixing technique is implemented. To use the large area inspection ability of the guided wave, the main research issue in this paper is focused on the guided wave mixing technique. Moreover, several bulk wave mixing theory equations become good concepts for guided wave mixing theoretical study, and the conventional nonlinear technique and guided wave mixing experimental results are compared in this study to confirm the reliability. This technique can play an important role in quantitatively discriminating fine damage by minimizing the nonlinearity of the equipment system

Ultrasonic Nonlinearity Experiment due to Plastic Deformation of Aluminum Plate Due to Bending Damage

The nonlinear ultrasonic evaluation technique is useful for assessing micro-defects and microstructure changes caused by fatigue or bending damage. In particular, the guided wave is advantageous for long-distance testing such as piping and plate. Despite these advantages, the study of nonlinear guided wave propagation has received relatively less attention compared to bulk wave techniques. Furthermore, there is a lack of research on the correlation between nonlinear parameters and material properties. In this study, the relationship between nonlinear parameters and plastic deformation resulting from bending damage was experimentally investigated using Lamb waves. The findings indicated an increase in the nonlinear parameter for the specimen, which was loaded within the elastic limit. Inversely, regions of maximum deflection in specimens with plastic deformation exhibited a decrease in the nonlinear parameter. This research is expected to be helpful for maintenance technology in the nuclear power plant and aerospace fields that require high reliability and accuracy

High-Precision Noncontact Guided Wave Tomographic Imaging of Plate Structures Using a DHB Algorithm

The safety diagnostic inspection of large plate structures, such as nuclear power plant containment liner plates and aircraft wings, is an important issue directly related to the safety of life. This research intends to present a more quantitative defect imaging in the structural health monitoring (SHM) technique by using a wide range of diagnostic techniques using guided ultrasound. A noncontact detection system was applied to compensate for such difficulties because direct access inspection is not possible for high-temperature and massive areas such as nuclear power plants and aircraft. Noncontact systems use unstable pulse laser and air-coupled transducers. Automatic detection systems were built to increase inspection speed and precision and the signal was measured. In addition, a new Difference Hilbert Back Projection (DHB) algorithm that can replace the reconstruction algorithm for the probabilistic inspection of damage (RAPID) algorithm used for imaging defects has been successfully applied to quantitative imaging of plate structure defects. Using an automatic detection system, the precision and detection efficiency of data collection has been greatly improved, and the same results can be obtained by reducing errors in experimental conditions that can occur in repeated experiments. Defects were made in two specimens, and comparative analysis was performed to see if each algorithm can quantitatively represent defects in multiple defects. The new DHB algorithm presented the possibility of observing and predicting the growth direction of defects through the continuous monitoring system

Eliminating murine norovirus, Helicobacter hepaticus, and intestinal protozoa by embryo transfer for an entire mouse barrier facility

Pathogens can affect physiological and immunological reactions in immunocompromised animals and genetically engineered mice. Specifically, murine norovirus (MNV), Helicobacter, and intestinal protozoa are prevalent in rodent laboratory facilities worldwide. In this study, microbiological test results of the soiled bedding of sentinel mice showed the prevalence of MNV (50.9%, 28/55), Helicobacter hepaticus (29.1%, 16/55), Trichomonas spp. (14.5%, 8/55), and Entamoeba spp. (32.7%, 18/55). No single infections were detected as all cases were confirmed to have complex infections with two or four pathogens. In previous studies, the success rate of the cross-fostering method was not perfect; therefore, in this study, the entire mouse strain of the SPF rodent facility was rederived using embryo transfer. For up to three years, we confirmed that the results were negative with regular health surveillance tests. Embryo transfer was, thus, determined to be an effective method for the rederivation of specific pathogen free (SPF) barrier mouse facilities. This is the report for the effectiveness of embryo transfer as an example of successful microbiological clean-up of a mouse colony with multiple infections in an entire SPF mouse facility and embryo transfer may be useful for rederiving.Y

Postnatal exposure to trimethyltin chloride induces retinal developmental neurotoxicity in mice via glutamate and its transporter related changes

Exposure to toxic substances during postnatal period is one of the major factors causing retinal developmental defects. The developmental toxicity of trimethyltin chloride (TMT), a byproduct of an organotin compound widely used in agriculture and industrial fields, has been reported; however, the effect on the mammalian retina during postnatal development and the mechanism have not been elucidated to date. We exposed 0.75 and 1.5 mg/kg of TMT to neonatal ICR mice (1:1 ratio of male and female) up to postnatal day 14 and performed analysis of the retina: histopathology, apoptosis, electrophysiological function, glutamate concentration, gene expression, and fluorescence immunostaining. Exposure to TMT caused delayed eye opening, eye growth defect and thinning of retinal layer. In addition, apoptosis occurred in the retina along with b-wave and spiking activity changes in the micro-electroretinogram. These changes were accompanied by an increase in the concentration of glutamate, upregulation of astrocyte-related genes, and increased expression of glial excitatory amino acid transporter (EAAT) 1 and 2. Conversely, EAAT 3, 4, and 5, mainly located in the neurons, were decreased. Our results are the first to prove postnatal retinal developmental neurotoxicity of TMT at the mammalian model and analyze the molecular, functional as well as morphological aspects to elucidate possible mechanisms: glutamate toxicity with EAAT expression changes. These mechanisms may suggest not only a strategy to treat but also a clue to prevent postnatal retina developmental toxicity of toxic substances. © 2023 Elsevier Inc.N

Flubendazole exposure disrupts neural development and function of zebrafish embryos (Danio rerio)

Flubendazole (FBZ) is a benzimidazole anthelmintic drug widely used for treating parasitic infections by disrupting microtubule formation and function through tubulin binding. Recently, its use has extended to include anticancer applications, leading to increased environmental exposure to benzimidazole drugs. However, the impact of FBZ on neural development in aquatic organisms, particularly in aquatic vertebrates, remains poorly understood. This study aimed to investigate the potential developmental toxicity of FBZ during neural development using zebrafish model. Various assessments, including analysis of overall developmental changes, morphological abnormalities, apoptosis, gene expression alterations, axon length measurements, and electrophysiological neural function, were performed. FBZ exposure resulted in concentration-dependent effects on survival rate, hatching rate, heartbeat, and the occurrence of developmental abnormalities. Notably, FBZ-induced changes included reductions in body length, head size, and eye size, as well as the detection of apoptotic cells in the central nervous system. Gene expression analysis revealed upregulation of apoptosis-related genes (p53, casp3, and casp8), downregulation of neural differentiation-related genes (shha, nrd, ngn1, and elavl3), and alterations in neural maturation and axon growth-related genes (gap43, mbp, and syn2a). Additionally, shortened motor neuron axon length and impaired electrophysiological neural function were observed. These findings provide novel insights into the potential risks of FBZ on the neural development of zebrafish embryos, emphasizing the need for risk prevention strategies and therapeutic approaches to address the environmental toxicity of benzimidazole anthelmintics.Y

Albendazole exerts antiproliferative effects on prostate cancer cells by inducing reactive oxygen species generation

Benzimidazole derivatives are used for their antihelmintic properties, but have also been reported to exert anticancer effects. In the present study, the anticancer effects of albendazole on prostate cancer cells were assessed using proliferation, clonogenic and migration assays. To investigate the anticancer mechanisms of albendazole, reactive oxygen species (ROS) levels were measured, and the expression of genes associated with oxidative stress and Wnt/beta-catenin signaling was confirmed by reverse transcription-quantitative PCR and western blotting. Albendazole selectively inhibited the proliferation of the PC3, DU145, LNCaP and AT2 prostate cancer cell lines at concentrations that did not affect the proliferation of a normal prostate cell line (RWPE-1). Albendazole also inhibited the colony formation and migration of PC3 and DU145 cells, as well as inducing ROS production. Diphenyleneiodonium chloride, an inhibitor of NADPH oxidase (NOX), one of the sources of ROS, decreased basal ROS levels in the PC3 and DU145 cells, but did not reduce albendazole-associated ROS production, suggesting that ROS production following albendazole treatment was NOX-independent. The anticancer effect was decreased when albendazole-induced ROS was reduced by treatment with antioxidants (glutathione and N-acetylcysteine). Furthermore, albendazole decreased the mRNA expression of CDGSH iron sulfur domain 2, which regulates antioxidant activity against ROS, as well as the antioxidant enzymes catalase, and glutathione peroxidase 1 and 3. Albendazole also decreased the mRNA expression of catenin beta 1 and transcription factor 4, which regulate Wnt/beta-catenin signaling and its associated targets, Twist family BHLH transcription factor 1 and BCL2. The albendazole-related decrease in the expression levels of oxidative stress-related genes and Wnt/beta-catenin signaling proteins was thought to be associated with ROS production. These results suggest that the antihelmintic drug, albendazole, has inhibitory effects against prostate cancer cells in vitro. Therefore, albendazole may potentially be used as a novel anticancer agent for prostate cancer.Y

Ultrasonic Assessment of Thickness and Bonding Quality of Coating Layer Based on Short-Time Fourier Transform and Convolutional Neural Networks

Ultrasonic non-destructive analysis is a promising and effective method for the inspection of protective coating materials. Offshore coating exhibits a high attenuation rate of ultrasonic energy due to the absorption and ultrasonic pulse echo testing becomes difficult due to the small amplitude of the second echo from the back wall of the coating layer. In order to address these problems, an advanced ultrasonic signal analysis has been proposed. An ultrasonic delay line was applied due to the high attenuation of the coating layer. A short-time Fourier transform (STFT) of the waveform was implemented to measure the thickness and state of bonding of coating materials. The thickness of the coating material was estimated by the projection of the STFT into the time-domain. The bonding and debonding of the coating layers were distinguished using the ratio of the STFT magnitude peaks of the two subsequent wave echoes. In addition, the advantage of the STFT-based approach is that it can accurately and quickly estimate the time of flight (TOF) of a signal even at low signal-to-noise ratios. Finally, a convolutional neural network (CNN) was applied to automatically determine the bonding state of the coatings. The time–frequency representation of the waveform was used as the input to the CNN. The experimental results demonstrated that the proposed method automatically determines the bonding state of the coatings with high accuracy. The present approach is more efficient compared to the method of estimating bonding state using attenuation

Loss of glutathione peroxidase 3 induces ROS and contributes to prostatic hyperplasia in Nkx3.1

Background Glutathione peroxidase 3 (Gpx3) protects cells from oxidative stress, and its reduced expression in human prostate cancer has been reported. Objectives We hypothesized thatGpx3might play an important role in the development of prostatic intraepithelial neoplasia (PIN), a pre-cancerous state of the prostate, and aimed to highlight the underlying molecular mechanism. Materials and Methods The following double-knockout miceNkx3.1-/-; Gpx3+/+, Nkx3.1-/-; Gpx3+/-, Nkx3.1-/-; Gpx3-/- were produced. Randomly divided animals were weighed, and their genitourinary tract (GUT) weights were determined after euthanasia at 4, 8, and 12 months. The mRNA expression of the genes involved in oxidative stress and Wnt signaling was analyzed in the prostate. Histopathology, ROS, and superoxide dismutase (SOD) activities were also measured. Results Loss of Gpx3 did not affect body weight and GUT weight in Nkx3.1 knockout mice. The mRNA expression of SOD3, iNOS, Hmox, and CISD2, which are associated with oxidative stress, was increased in Nkx3.1-/-; Gpx3-/- mice at 4 months but decreased at 8 and 12 months. There was no change in beta-catenin and its targets associated with Wnt signaling. Increased ROS and decreased SOD activity were observed in Nkx3.1-/-; Gpx3-/- mice at 12 months of age. The histopathologic score and epithelium thickness were increased, and lumen area was decreased in Gpx3 knockout mice. Discussion and Conclusions Gpx3loss increased the hyperplasia of PIN in the pre-cancerous stage of the prostate. Loss ofGpx3induced oxidative stress. Histopathologically, no invasive carcinoma was identified, andGpx3loss did not increase Wnt/beta-catenin signaling. Further research on the role of GPX3 in the transition of PIN to invasive carcinoma is needed. We show, for the first time, that the antioxidant enzyme GPX3 plays a vital role in inhibiting hyperplasia in the PIN stage of the prostate gland in vivo.N