Preliminary report of Mycoplasma Wenoynii and Candidatus Mycoplasma haemobos infection in Korean native cattle.

BACKGROUND: Hemotropic mycoplasmas or hemoplasmas are bacteria that attach to the erythrocyte surface and cause bovine hemoplasmosis. Two species, Mycoplasma wenyonii and Candidatus Mycoplasma haemobos, have been identified and shown to be distributed worldwide. However, there is currently no information available on hemoplasmas in cattle in the Republic of Korea. The aim of this study was to investigate the presence of hemoplasmas in Korean native cattle and to evaluate the association between hemoplasma infection and anemia. METHODS: One farm was selected, at which blood samples were collected from 104 Korean native cattle [grazing cattle (n = 89) and housed cattle (n = 15)]. Hemoplasmas were detected via polymerase chain reaction analysis and complete blood counts were also performed. RESULTS: The overall prevalence of hemoplasmas was 34% (35/104); 20.2% (21/104) for M. wenyonii, 3.8% (4/104) for C. M. haemobos, and 9.6% (10/104) for co-infection. Candidatus Mycoplasma haemobos was detected only in grazing cattle. Of red blood cell (RBC) parameters, C. M. haemobos-infected cattle had lower RBC and hematocrit, and higher mean cell volume than hemoplasma-negative cattle, although none of these differences were statistically significant. This is the first study to report the occurrence of M. wenyonii and C. M. haemobos. Mycoplasma wenyonii is more prevalent than C. M. haemobos in Korean native cattle. The results did not show an association between hemoplasma infection and anemia. CONCLUSIONS: Considering the infection rate of hemoplasmas shown in this study, further studies, such as on the pathogenicity and clinical significance of hemoplasmas are necessary.

Genotype Analysis of Respiratory Syncytial Virus Before and After the COVID-19 Pandemic Using Whole-Genome Sequencing: A Prospective, Single-Center Study in Korea From 2019 to 2022.

BACKGROUND: Respiratory syncytial virus (RSV), a highly transmissible virus, is the leading cause of lower respiratory tract infections. We examined molecular changes in the RSV genome before and after the coronavirus disease 2019 (COVID-19) pandemic in Korea, and investigated whether drug-resistant mutations were present. METHODS: In this prospective, single-center study, RSV-positive respiratory samples were collected between September 2019 and December 2022. Long-read whole-genome sequencing (WGS) was performed, and the presence of known drug-resistant substitutions for palivizumab, nirsevimab, and suptavumab was investigated. RESULTS: Overall, 288 respiratory samples were collected from 276 children. WGS data were available for 133 samples (71 and 62 samples from the pre- and post-pandemic periods, respectively). All RSV-A strains (n = 56) belonged to the GA2.3.5 (ON1) genotype, whereas all RSV-B strains (n = 77) belonged to the GB5.0.5a (BA) genotype. No significant differences in genotypes were observed between the pre- and post-pandemic periods. In addition, no notable mutations related to nirsevimab or palivizumab resistance were detected in the F gene. However, the L172Q and S173L substitutions, which are known to confer resistance to suptavumab, were present in all RSV-B samples. CONCLUSION: Despite the unprecedented interruption of RSV seasonality, there were no significant molecular changes in circulating RSV strains in Korea related to nirsevimab or palivizumab resistance before and after the COVID-19 pandemic. However, RSV-specific drug-resistance substitutions for suptavumab were identified.

Sensors and Sensing Devices Utilizing Electrorheological Fluids and Magnetorheological Materials-A Review.

This paper comprehensively reviews sensors and sensing devices developed or/and proposed so far utilizing two smart materials: electrorheological fluids (ERFs) and magnetorheological materials (MRMs) whose rheological characteristics such as stiffness and damping can be controlled by external stimuli; an electrical voltage for ERFs and a magnetic field for MRMs, respectively. In this review article, the MRMs are classified into magnetorheological fluids (MRF), magnetorheological elastomers (MRE) and magnetorheological plastomers (MRP). To easily understand the history of sensing research using these two smart materials, the order of this review article is organized in a chronological manner of ERF sensors, MRF sensors, MRE sensors and MRP sensors. Among many sensors fabricated from each smart material, one or two sensors or sensing devices are adopted to discuss the sensing configuration, working principle and specifications such as accuracy and sensitivity. Some sensors adopted in this article include force sensors, tactile devices, strain sensors, wearable bending sensors, magnetometers, display devices and flux measurement sensors. After briefly describing what has been reviewed in a conclusion, several challenging future works, which should be undertaken for the practical applications of sensors or/and sensing devices, are discussed in terms of response time and new technologies integrating with artificial intelligence neural networks in which several parameters affecting the sensor signals can be precisely and optimally tuned. It is sure that this review article is very helpful to potential readers who are interested in creative sensors using not only the proposed smart materials but also different types of smart materials such as shape memory alloys and active polymers.

A Novel Tactile Sensing System Utilizing Magnetorheological Structures for Dynamic Contraction and Relaxation Motions.

It is well known that the rheological properties of magnetorheological (MR) material change under a magnetic field. So far, most works on MR materials have been oriented toward actuating characteristics instead of sensing functions. In this work, to realize dynamic tactile motion, a spherical MR structure was designed as a sensor, incorporating a magnetic circuit core to provide maximum dynamic motion. After manufacturing a prototype (sample), a sinusoidal magnetic field of varying exciting frequency and magnitude was applied to the sample, and the dynamic contraction and relaxation motion depending on the exciting magnetic field was observed. Among the test results, when 10% deformation occurred, the instantaneous force generated was from 2.8 N to 8.8 N, and the force when relaxed was from 1.2 N to 3.5 N. It is also shown that the repulsive force within this range can be implemented using an acceptable input current. The special tactile sensing structure proposed in this work can be used as a sensor to measure the field-dependent viscoelastic properties of human tissues such as stomach, liver, and overall body. In addition, it could be usefully applied to robot surgery, because it can mimic the dynamic motions of various human organs under various surgical conditions.

Antimicrobial properties of Pediococcus acidilactici and Pediococcus pentosaceus isolated from silage.

AIMS: The objective of this study was to isolate multifunctional bacteriocin-producing strains; to characterize the expressed bacteriocin for the control of Listeria monocytogenes and vancomycin-resistant Enterococcus; to evaluate the safety of studied strains; and to explore their antifungal activity. METHODS AND RESULTS: Two Pediococcus strains were isolated from silage samples obtained from an organic farm in Belogradchik, Bulgaria. The strains were identified by 16S rRNA sequencing analysis and characterized as bacteriocins producers. Strong antimicrobial activity was detected against more than 74 different strains of Listeria monocytogenes, 27 different vancomycin-resistant Enterococcus strains. In addition, studied strains were able to inhibit the growth of strains of Alternaria alternate, Aspergillus flavus, Aspergillus niger, Cladosporium sphaerospermum, Penicillium chrysogenum and Penicillium expansum. Some aspects of the antimicrobial mode of action were evaluated, including killing curves and aggregation properties. Both strains generated positive PCR results for the presence of pediocin PA-1, but not for other bacteriocins evaluated in this screening process. Metabolomic analysis of the cell-free supernatants from both strains was performed in order to explain the observed antifungal activity against different moulds. According to PCA and PLS-DA score plot, P. acidilactici ST3522BG and P. pentosaceus ST3633BG were clearly clustered from control (MRS). Increases in the production of benzoic acid, 2-hydroxyisocaproic acid, ß-phenyl-lactic acid, α-hydroxybutyric acid and 1,3-butanediol were recorded, these metabolites were previously described as antifungal. CONCLUSIONS: Pediococcus acidilactici ST3522BG and P. pentosaceus ST3633BG were evaluated as producing bacteriocin strains with high specificity against Listeria and vancomycin-resistant Enterococcus species. In addition, both investigated Pediococcus strains were evaluated as producer of effective antifungal metabolites with potential for the inhibition of mycotoxin-producing moulds. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this report is a pioneer in the evaluation of Pediococcus strains isolated from silage with highly specific bacteriocinogenic antimicrobial activity against Listeria spp. and vancomycin-resistant Enterococcus spp., and antifungal activity against mycotoxin-producing moulds.

Characterization and safety evaluation of two beneficial, enterocin-producing Enterococcus faecium strains isolated from kimchi, a Korean fermented cabbage.

Enterococcus faecium ST20Kc and ST41Kc were isolated from kimchi, a traditional Korean fermented cabbage. Bacteriocins produced by both strains exhibited strong activity against Listeria monocytogenes and various Enterococcus spp., including 30 vancomycin-resistant enterococcal strains, but not against other lactic acid bacteria (LAB) on the evaluated test panel. The antimicrobials produced by the strains were found to be proteinaceous and stable even after exposure to varying pH, temperature, and chemicals used in the industry and laboratory processes. Antimicrobial activity of both strains was evaluated as bactericidal against exponentially growing cultures of L. monocytogenes ATCC® 15313™ and Enterococcus faecalis 200A. Based on tricine-SDS-PAGE, the molecular weights of the bacteriocins produced by the strains were between 4 and 6 kDa. Additionally, both strains were susceptible to antibiotics, including vancomycin, kanamycin, gentamycin, ampicillin, streptomycin, tylosin, chloramphenicol, clindamycin, and tetracycline. Adhesion genes, map, mub, and EF-Tu, were also detected in the genomes of both strains. With gastrointestinal stress induction, both strains showed high individual survival rates, and capability to reduce viable counts of L. monocytogenes ATCC® 15313™ and Enterococcus faecalis 200A in mixed cultures. Based on the metabolomics analysis, both strains were found to produce additional antimicrobial compounds, particularly, lactic acid, phenyllactic acid, and phenethylamine, which can be potentially involved in the antimicrobial interaction with pathogenic microorganisms.

Secured and Deterministic Closed-Loop IoT System Architecture for Sensor and Actuator Networks.

Sensors, actuators, and wireless communication technologies have developed significantly. Consequently, closed-loop systems that can be monitored and controlled by devices in IoT environments, such as farms and factories, have emerged. Such systems are realized by means of cloud-level and edge-level implementations. Among them, with a model that generates real-time control decisions at the cloud level, it might be difficult to ensure the determinism of real-time control due to communication overheads. In addition, if the actuator is remotely controlled at the cloud level, it is difficult to secure control safety against external hacking or device malfunction. Herein, we propose a system architecture that can fulfil real-time performance and safety requirements with two commonly used devices, Field Edge Unit (FEU) and Current Sensing Tag (CST), in a closed-loop IoT environment. By using these devices, we designed a special architecture that can be commonly used in various closed-loop sensing and actuating applications. In this study, the proposed architecture is evaluated by applying it to a smart farm application.

A Cylindrical Grip Type of Tactile Device Using Magneto-Responsive Materials Integrated with Surgical Robot Console: Design and Analysis.

This paper proposes a cylindrical grip type of tactile device that is effectively integrated to a surgical robot console so that a surgeon can easily touch and feel the same stiffness as the operating organs. This is possible since the yield stress (or stiffness) of magnetic-responsive materials can be tuned or controlled by the magnetic field intensity. The proposed tactile device consists of two main parts: a magnetorheological elastomer (MRE) layer and a magnetorheological fluid (MRF) core. The grip shape of the device to be positioned on the handle part of the master of the surgical robot is configured and its operating principle is discussed. Then, a couple of equations to calculate the stiffness from the gripping force and the field-dependent yield stress of MRF are derived and integrated using the finite element analysis (FEA) model. After simulating the stiffness of the proposed tactile device as a function of the magnetic field intensity (or current), the stiffnesses of various human organs, including the liver and heart, are calculated from known data of an elastic modulus. It is demonstrated from comparative data between calculated stiffness from human tissues and simulated stiffness from FEA that the proposed tactile device can generate sufficient stiffness with a low current level to recognize various human organs which are significantly required in the surgical robot system.

Assessment of the safety and anti-inflammatory effects of three Bacillus strains in the respiratory tract.

Chronic respiratory diseases are part of accumulating health problems partly due to worldwide increase in air pollution. By their antimicrobial and immunomodulatory properties, some probiotics constitute promising alternatives for the prevention and treatment of chronic respiratory diseases. We have isolated Bacillus strains from Korean fermented foods and selected three potentially probiotic strains (two Bacillus subtilis and one Bacillus amyloliquefaciens) based on safety, antimicrobial efficacy, activity against airborne pathogens and their immunomodulatory properties in vivo. Safety evaluation included in silico analysis for confirming absence of virulence genes. Safety for the respiratory tract was confirmed by an in vivo pathogenicity test using a murine model. Antimicrobial activity was displayed against several airborne pathogens. Potential antimicrobial metabolites such as 2,3-butanediol and propylene glycol were identified as possible antagonistic agents. Immunomodulatory properties in vitro were confirmed by upregulation of IL-10 expression in a macrophage cell line. Intranasal instillation and inhalation in an ovalbumin (OVA)-induced lung inflammation murine model reduced T helper type 2 (Th2) cytokines at transcriptional and protein levels in the lungs. The safety and potentially beneficial role of these Bacillus strains could be demonstrated for the respiratory tract of a murine model.

Chimerism Assay Using Single Nucleotide Polymorphisms Adjacent and in Linkage-Disequilibrium Enables Sensitive Disease Relapse Monitoring after Hematopoietic Stem-Cell Transplantation.

BACKGROUND: Short tandem repeat (STR)-based chimerism analysis has been widely used for chimerism monitoring after hematopoietic stem-cell transplantation (HSCT), but technical artifacts can be problematic. We designed a chimerism assay using single nucleotide polymorphisms (SNPs) adjacent and in linkage-disequilibrium (CASAL), which doubly checked for SNP pairs, and thus could reduce background errors and increase analytical sensitivity. METHODS: CASAL targeted 84 SNP pairs within 10 bp distance and in perfect linkage-disequilibrium. Using undiluted and serially diluted samples, baseline error rates, and linearity was calculated. Clinical performance of CASAL was evaluated in comparison with a conventional STR assay, using 191 posttransplant samples from 42 patients with HSCT. RESULTS: CASAL had ∼10 times lower baseline error rates compared to that of ordinary next-generation sequencing. Limit of detection and quantification of CASAL were estimated to be 0.09 and 0.39%, respectively, with a linear range of 0.1-100%. CASAL correlated well with STR assay (r2 = 0.99) and the higher sensitivity enabled detection of low-level recipient chimerism and earlier prediction of relapse. CONCLUSIONS: CASAL is a simple, analytically sensitive and accurate assay that can be used in clinical samples after HSCT with a higher performance compared to that of traditional assays. It should also be useful in other forensic and archeological testing.

A New Tactile Transfer Cell Using Magnetorheological Materials for Robot-Assisted Minimally Invasive Surgery.

This paper proposes a new type of tactile transfer cell which can be effectively applied to robot-assisted minimally invasive surgery (RMIS). The proposed tactile device is manufactured from two smart materials, a magnetorheological fluid (MRF) and a magnetorheological elastomer (MRE), whose viscoelastic properties are controllable by an external magnetic field. Thus, it can produce field-dependent repulsive forces which are equivalent to several human organs (or tissues) such as a heart. As a first step, an appropriate tactile sample is made using both MRF and MRE associated with porous foam. Then, the microstructures of these materials taken from Scanning Electron Microscope (SEM) images are presented, showing the particle distribution with and without the magnetic field. Subsequently, the field-dependent repulsive force of the sample, which is equivalent to the stress relaxation property of viscoelastic materials, are measured at several compressive deformation depths. Then, the measured values are compared with the calculated values obtained from Young's modulus of human tissue data via the finite element method. It is identified from this comparison that the proposed tactile transfer cell can mimic the repulsive force (or hardness) of several human organs. This directly indicates that the proposed MR materials-based tactile transfer cell (MRTTC in short) can be effectively applied to RMIS in which the surgeon can feel the strength or softness of the human organ by just changing the magnetic field intensity. In this work, to reflect a more practical feasibility, a psychophysical test is also carried out using 20 volunteers, and the results are analyzed, presenting the standard deviation.

Self-Organizing IoT Device-Based Smart Diagnosing Assistance System for Activities of Daily Living.

Activity of daily living (ADL) is a criterion for evaluating the performance ability of daily life by recognizing various activity events occurring in real life. However, most of the data necessary for ADL evaluation are collected only through observation and questionnaire by the patient or the patient's caregiver. Recently, Internet of Things(IoT) device studies using various environmental sensors are being used for ADL collection and analysis. In this paper, we propose an IoT Device Platform for ADL capability measurement. Wearable devices and stationary devices recognize activity events in real environments and perform user identification through various sensors. The user's ADL data are sent to the network hub for analysis. The proposed IoT platform devices support many sensor devices such as acceleration, flame, temperature, and humidity in order to recognize various activities in real life. In addition, in this paper, using the implemented platform, ADL measurement test was performed on hospital patients. Through this test, the accuracy and reliability of the platform are analyzed.

FIRST REPORT OF ENTEROCYTOZOON BIENEUSI FROM AN AFRICAN LION (PANTHERA LEO) IN A ZOO IN THE REPUBLIC OF KOREA.

Enterocytozoon bieneusi is the most common species of microsporidia that infects humans and animals worldwide. However, no information is available on E. bieneusi infection among zoo animals in the Republic of Korea (ROK). Here, we investigated the prevalence of E. bieneusi among animals kept in zoos and the zoonotic potential of the E. bieneusi identified. E. bieneusi was detected only in one African lion (Panthera leo) with diarrhea, using PCR and sequencing analysis of the internal transcribed spacer (ITS) of the rRNA gene. A phylogenetic analysis based on the ITS gene showed that the lion isolate was classified into a novel genotype KPL belonging to Group 2. The KPL genotype identified in this study differed from genotype I in 6 nucleotides and from genotype I-like in 3 nucleotides, respectively, indicating that Group 2 has the capacity to infect a wide range of hosts. This is the first report of the presence of E. bieneusi in an African lion housed in a zoo in the ROK. Further investigation is necessary to study E. bieneusi infection among zoo animals in various regions and to determine the transmission route, in order to control E. bieneusi infection.

Human induced pluripotent stem cell line banking for the production of rare blood type erythrocytes.

BACKGROUND: The in vitro production of mature human red blood cells (RBCs) from induced pluripotent stem cells (iPSCs) has been the focus of research to meet the high demand for blood transfusions. However, limitations like high costs and technological requirements restrict the use of RBCs produced by iPSC differentiation to specific circumstances, such as for patients with rare blood types or alloimmunized patients. In this study, we developed a detailed protocol for the generation of iPSC lines derived from peripheral blood of donors with O D-positive blood and rare blood types (D-and Jr(a-)) and subsequent erythroid differentiation. METHODS: Mononuclear cells separated from the peripheral blood of O D-positive and rare blood type donors were cultured to produce and expand erythroid progenitors and reprogrammed into iPSCs. A 31-day serum-free, xeno-free erythroid differentiation protocol was used to generate reticulocytes. The stability of iPSC lines was confirmed with chromosomal analysis and RT-PCR. Morphology and cell counts were determined by microscopy observations and flow cytometry. RESULTS: Cells from all donors were successfully used to generate iPSC lines, which were differentiated into erythroid precursors without any apparent chromosomal mutations. This differentiation protocol resulted in moderate erythrocyte yield per iPSC. CONCLUSIONS: It has previously only been hypothesized that erythroid differentiation from iPSCs could be used to produce RBCs for transfusion to patients with rare blood types or who have been alloimmunized. Our results demonstrate the feasibility of producing autologous iPSC-differentiated RBCs for clinical transfusions in patients without alternative options.

Sphingobium terrigena sp. nov., isolated from gasoline-contaminated soil.

A Gram-stain-negative, strictly aerobic bacterial strain, designated EO9T, was isolated from gasoline-contaminated soil in the Republic of Korea. Cells were non-motile short rods showing catalase- and oxidase-positive reactions. Growth was observed at 10-37 °C (optimum, 30 °C), at pH 6.0-9.0 (pH 6.5) and in the presence of 0-0.5 % (w/v) NaCl (0 %). Ubiquinone-10 (Q-10) and spermidine were identified as the predominant respiratory quinone and polyamine, respectively. Summed feature 8 (comprising C18:1ω7c/C18:1ω6c), summed feature 3 (comprising C16:1ω7c/C16:1ω6c), C16:0 and C14:0 2-OH were identified as major cellular fatty acids. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, sphingoglycolipid, phosphatidylglycerol and an unidentified phospholipid. The G+C content of the genomic DNA was 62.8 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain EO9T formed a tight phylogenetic lineage with Sphingobium xenophagum NBRC 107872T and Sphingobium hydrophobicum C1T within the genus Sphingobium. Strain EO9T was most closely related to S. xenophagum NBRC 107872T (97.2 %) and S. hydrophobicum C1T (97.2 %), but DNA-DNA relatedness levels between strain EO9T and the type strains of S. xenophagum and S. hydrophobicum were 37.1 and 36.8 % , respectively. Based on its phenotypic, chemotaxonomic and molecular features, strain EO9T clearly represents a novel species of the genus Sphingobium, for which the name Sphingobium terrigena sp. nov. is proposed. The type strain is EO9T (=KACC 19523T=JCM 32762T).

Integrated Metabolomics and Transcriptomics Unravel the Metabolic Pathway Variations for Different Sized Beech Mushrooms.

Beech mushrooms (Hypsizygus marmoreus) are largely relished for their characteristic earthy flavor, chewy-texture, and gustatory and nutritional properties in East Asian societies. Intriguingly, the aforementioned properties of beech mushroom can be subsumed under its elusive metabolome and subtle transcriptome regulating its various stages of growth and development. Herein, we carried out an integrated metabolomic and transcriptomic profiling for different sized beech mushrooms across spatial components (cap and stipe) to delineate their signature pathways. We observed that metabolite profiles and differentially expressed gene (DEGs) displayed marked synergy for specific signature pathways according to mushroom sizes. Notably, the amino acid, nucleotide, and terpenoid metabolism-related metabolites and genes were more abundant in small-sized mushrooms. On the other hand, the relative levels of carbohydrates and TCA intermediate metabolites as well as corresponding genes were linearly increased with mushroom size. However, the composition of flavor-related metabolites was varying in different sized beech mushrooms. Our study explores the signature pathways associated with growth, development, nutritional, functional and organoleptic properties of different sized beech mushrooms.

Nocardioides currus sp. nov., isolated from a mobile car air-conditioning system.

A strictly aerobic Gram-stain-positive bacterial strain, designated IB-3T, was isolated from a car air-conditioning system in the Republic of Korea. Cells were non-motile rods showing catalase- and oxidase-positive reactions. Growth of IB-3T was observed at 20-40 °C (optimum, 25 °C), at pH 6.5-9.0 (optimum, pH 7.5) and in the presence of 0-1 % (w/v) NaCl (optimum, 0 %). Menaquinone-8 (H4) was detected as the predominant respiratory quinone and iso-C16 : 0, 10-methyl-C17 : 0, iso-C17 : 0, C18 : 1ω9c, C17 : 1ω8c, C18 : 0, 10-methyl-C18 : 0 (TBSA) and C17 : 0 were identified as the major cellular fatty acids. Phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and phosphatidylinositol were detected as the major polar lipids. The major cell wall peptidoglycan type was ll-2,6-diaminopimelic acid. The G+C content of the genomic DNA was 71.5 mol%. IB-3T was most closely related to Nocardioides terrigenaDS-17T with a 98.0 % 16S rRNA gene sequence similarity. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that IB-3T formed a distinct phylogenetic lineage within the genus Nocardioidesof the family Nocardioidaceae. On the basis of the phenotypic, chemotaxonomic and molecular features, IB-3T represents a novel species of the genus Nocardioides, for which the name Nocardioidescurrus sp. nov. is proposed. The type strain is IB-3T (=KACC 19522T=JCM 32672T).

Evaluation of subchronic exposure to triclosan on hepatorenal and reproductive toxicities in prepubertal male rats.

Triclosan (TCS), a common antimicrobial ingredient, is present in many consumer products, including soaps, shampoos, and toothpaste. Owing to its widespread use, potential adverse effects on animals and humans may arise from lifetime exposure, but data on chronic prepubertal exposure of TCS are still lacking. The aim of the present study was to investigate the influence of subchronic TCS exposure (0.25, 25, 250, or 750 mg/kg) on target organ toxicity in prepubertal male rats. After daily administration of TCS to rats by oral gavage for 60 d, a significant reduction in body weight and relative weights of liver, kidneys, testes, and adrenal glands was observed in the 750-mg/kg (high dose) group. Serum alanine aminotransferase and aspartate aminotransferase activities as well as levels of blood urea nitrogen, and creatinine were significantly increased at 750 mg/kg TCS. Further, TCS (750 mg/kg) elevated the protein expressions of hepatic CYP2B1, RXR/PPAR, and levels of malondialdehyde. High-dose TCS exposure induced histological changes as evidenced by reduction of Bowman's space, occlusion of the tubular lumen, and degeneration of tubular epithelial cells in the kidney. Tubular necrosis was confirmed as evidenced by a rise in expression of high mobility group box 1 renal protein. Daily sperm production was significantly diminished by high doses of TCS with marked inhibition of androgen receptor protein expression. Our results indicated that subchronic exposure to excessively high concentrations of 750 mg/kg TCS induced hepatorenal and reproductive toxicities in prepubertal male rats; however, the biological relevance of these findings is questionable as these drug levels are not encountered in the environment.

A New Synthetic Histone Deacetylase Inhibitor, MHY2256, Induces Apoptosis and Autophagy Cell Death in Endometrial Cancer Cells via p53 Acetylation.

We previously discovered a novel sirtuin (SIRT) inhibitor, MHY2256, that exerts anticancer activity through p53 acetylation in MCF-7 human breast cancer cells. We investigated the anticancer activity of MHY2256 against hormone-related cancer, an endometrial cancer with a poor prognosis. The IC50 values of MHY2256 were shown to be much lower than those of salermide, a well-known SIRT inhibitor. Furthermore, MHY2256 significantly reduced the protein expression and activities of SIRT1, 2, and 3, with similar effects to salermide. Particularly, MHY2256 markedly inhibited tumor growth in a tumor xenograft mouse model of Ishikawa cancer cells. During the experimental period, there was no significant change in the body weight of mice treated with MHY2256. A detailed analysis of the sensitization mechanisms of Ishikawa cells revealed that late apoptosis was largely increased by MHY2256. Additionally, MHY2256 increased G1 arrest and reduced the number of cell cyclic-related proteins, suggesting that apoptosis by MHY2256 was achieved by cellular arrest. Particularly, p21 was greatly increased by MHY225656, suggesting that cell cycle arrest by p21 is a major factor in MHY2256 sensitization in Ishikawa cells. We also detected a significant increase in acetylated p53, a target protein of SIRT1, in Ishikawa cells after MHY2256 treatment. In a mouse xenograft model, MHY2256 significantly reduced tumor growth and weight without apparent side effects. These results suggest that MHY2256 exerts its anticancer activity through p53 acetylation in endometrial cancer and can be used for targeting hormone-related cancers.

Assessing translational efficiency by a reporter protein co-expressed in a cell-free synthesis system.

We demonstrate the use of a cell-free protein synthesis system as a convenient tool for assessing the relative translational efficiencies of genes. When sfGFP was used as a common reporter gene and co-expressed with a series of target genes, the intensities of sfGFP fluorescence from the co-expression reactions were highly correlated with the individual expression levels of the co-expressed genes. The relative translational efficiencies of genes estimated by this method were reproducible when the same genes were expressed in transformed Escherichia coli, suggesting that this method could be used as a universal tool for prognostic assessment of translational efficiency.