Detection and Characterization of Methylated Circulating Tumor DNA in Gastric Cancer.

Gastric cancer is the fifth most common disease in the world and the fourth most common cause of death. It is diagnosed through esophagogastroduodenoscopy with biopsy; however, there are limitations in finding lesions in the early stages. Recently, research has been actively conducted to use liquid biopsy to diagnose various cancers, including gastric cancer. Various substances derived from cancer are reflected in the blood. By analyzing these substances, it was expected that not only the presence or absence of cancer but also the type of cancer can be diagnosed. However, the amount of these substances is extremely small, and even these have various variables depending on the characteristics of the individual or the characteristics of the cancer. To overcome these, we collected methylated DNA fragments using MeDIP and compared them with normal plasma to characterize gastric cancer tissue or patients' plasma. We attempted to diagnose gastric cancer using the characteristics of cancer reflected in the blood through the cancer tissue and patients' plasma. As a result, we confirmed that the consistency of common methylated fragments between tissue and plasma was approximately 41.2% and we found the possibility of diagnosing and characterizing cancer using the characteristics of the fragments through SFR and 5'end-motif analysis.

Epigenetic Regulation of the Expression of T Cell Stimulatory and Inhibitory Factors by Histone H3 Lysine Modification Enzymes and Its Prognostic Roles in Glioblastoma.

BACKGROUND: This study aimed to identify the specific T cell co-stimulatory and co-inhibitory factors that play prognostic roles in patients with glioblastoma. Additionally, the unique histone H3 modification enzymes that regulate the expression levels of these specific co-stimulatory and co-inhibitory factors were investigated. METHODS: The medical records of 84 patients newly diagnosed with glioblastoma at our institution from January 2006 to December 2020 were retrospectively reviewed. Immunohistochemical (IHC) staining for T cell co-stimulatory factors (CD27, CD28, CD137, OX40, and ICOS), T cell co-inhibitory factors (CTLA4, PD1, PD-L1, TIM3, and CD200R), and histone H3 lysine modification enzymes (MLL4, RIZ, EZH1, NSD2, KDM5c, JMJD1a, UTX, and JMJD5) was performed on archived paraffin-embedded tissues obtained by biopsy or resection. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed for specific factors, which demonstrated causal relationships, in order to validate the findings of the IHC examinations. RESULTS: The mean follow-up duration was 27.5 months (range, 4.1-43.5 months). During this period, 76 patients (90.5%) died, and the mean OS was 19.4 months (95% confidence interval, 16.3-20.9 months). Linear positive correlations were observed between the expression levels of CD28 and JMJD1a (R2 linear = 0.982) and those of CD137 and UTX (R2 linear = 1.528). Alternatively, significant negative correlations were observed between the expression levels of CTLA4 and RIZ (R2 linear = -1.746) and those of PD-L1 and EZH1 (R2 linear = -2.118); these relationships were confirmed by qRT-PCR. In the multivariate analysis, increased expression levels of CD28 (P = 0.042), and CD137 (P = 0.009), and decreased expression levels of CTLA4 (P = 0.003), PD-L1 (P = 0.020), and EZH1 (P = 0.040) were significantly associated with longer survival. CONCLUSION: These findings suggest that the expression of certain T cell co-stimulatory factors, such as CD28 and CD 137, and co-inhibitory factors, such as CTLA4 and PD-L1 are associated with prognosis of glioblastoma patients.

Automatic Separation and Collection of Cancer-Related Substances from Clinical Samples.

Recently, liquid biopsies have been used to diagnose various diseases, including cancer. Body fluids contain many substances, including cells, proteins, and nucleic acids originating from normal tissues, but some of these substances also originate from the diseased area. The investigation and analysis of these substances in the body fluids play a pivotal role in the diagnosis of various diseases. Therefore, it is important to accurately separate the required substances, and several techniques are developed to be used for this purpose. We have developed a lab-on-a-disc type of device and platform named CD-PRIME. This device is automated and has good results for sample contamination and sample stability. Moreover, it has advantages of a good acquisition yield, a short operation time, and high reproducibility. In addition, depending on the type of disc to be mounted, plasma containing cell-free DNA, circulating tumor cells, peripheral blood mononuclear cells, or buffy coats can be separated. Thus, the acquisition of a variety of materials present in the body fluids can be done for a variety of downstream applications, including the study of omics.

A method for early diagnosis of lung cancer from tumor originated DNA fragments using plasma cfDNA methylome and fragmentome profiles.

Early detection is critical for minimizing mortality from cancer. Plasma cell-free DNA (cfDNA) contains the signatures of tumor DNA, allowing us to quantify the signature and diagnose early-stage tumors. Here, we report a novel tumor fragment quantification method, TOF (Tumor Originated Fragment) for the diagnosis of lung cancer by quantifying and analyzing both the plasma cfDNA methylation patterns and fragmentomic signatures. TOF utilizes the amount of ctDNA predicted from the methylation density information of each cfDNA read mapped on 6243 lung-tumor-specific CpG markers. The 6243 tumor-specific markers were derived from lung tumor tissues by comparing them with corresponding normal tissues and healthy blood from public methylation data. TOF also utilizes two cfDNA fragmentomic signatures: 1) the short fragment ratio, and 2) the 5' end-motif profile. We used 298 plasma samples to analyze cfDNA signatures using enzymatic methyl-sequencing data from 201 lung cancer patients and 97 healthy controls. The TOF score showed 0.98 of the area under the curve in correctly classifying lung cancer from normal samples. The TOF score resolution was high enough to clearly differentiate even the early-stage non-small cell lung cancer patients from the healthy controls. The same was true for small cell lung cancer patients.

Effects of Filler Functionalization on Filler-Embedded Natural Rubber/Ethylene-Propylene-Diene Monomer Composites.

Natural rubber (NR) presents a number of advantages over other types of rubber but has poor resistance to chemicals and aging. The incorporation of ethylene propylene diene monomer (EPDM) into the NR matrix may be able to address this issue. Mineral fillers, such as carbon black (CB) and silica are routinely incorporated into various elastomers owing to their low cost, enhanced processability, good functionality, and high resistance to chemicals and aging. Other fillers have been examined as potential alternatives to CB and silica. In this study, phlogopite was surface-modified using 10 phr of compatibilizers, such as aminopropyltriethoxysilane (A1S), aminoethylaminopropyltrimethoxysilane (A2S), or 3-glycidoxypropyltrimethoxysilane (ES), and mixed with NR/EPDM blends. The effects of untreated and surface-treated phlogopite on the mechanical properties of the rubber blend were then compared with those of common fillers (CB and silica) for rubbers. The incorporation of surface-modified phlogopite into NR/EPDM considerably enhanced various properties. The functionalization of the phlogopite surface using silane-based matters (amino- and epoxide-functionalized) led to excellent compatibility between the rubber matrix and phlogopite, thereby improving diverse properties of the elastomeric composites, with effects analogous to those of CB. The tensile strength and elongation at break of the phlogopite-embedded NR/EPDM composite were lower than those of the CB-incorporated NR/EPDM composite by 30% and 10%, respectively. Among the prepared samples, the ES-functionalized phlogopite showed the best compatibility with the rubber matrix, exhibiting a tensile strength and modulus of composites that were 35% and 18% higher, respectively, compared with those of the untreated phlogopite-incorporated NR/EPDM composite. The ES-functionalized phlogopite/NR/EPDM showed similar strength and higher modulus (by 18%) to the CB/NR/EPDM rubber composite, despite slightly lower elongation at break and toughness. The results of rebound resilience and compression set tests indicated that the elasticity of the surface-modified phlogopite/NR/EPDM rubber composite was higher than that of the silica- and CB-reinforced composites. These improvements could be attributed to enhancements in the physical and chemical interactions among the rubber matrix, stearic acid, and functionalized (compatibilized) phlogopite. Therefore, the functionalized phlogopite can be utilized in a wide range of applications for rubber compounding.

Selection of a Suitable Solvent Additive for 2-Methoxyethanol-Based Antisolvent-Free Perovskite Film Fabrication.

In this study, we compare various solvents with different Gutmann donor numbers as additives to improve the quality of perovskite films via an antisolvent-free process, using 2-methoxyethanol (2MOE) as the solvent. In 2MOE-based solutions, we found that the higher the donor number of the solvent, the lower the amount of the solvent required to form insoluble adducts. Furthermore, we found that N-methyl-2-pyrrolidone (NMP), which has a relatively low donor number and vapor pressure, can be added without a limitation to precipitation, while the degree of the intermediate phase in the as-deposited film is controlled by the amount of NMP added. We obtained pinhole-free and planar perovskite films by optimizing the amount of NMP added and fabricated devices based on NMP-assisted MAPbI3 and MAPbI3-xClx films, with efficiencies of 18.80 and 20.39%, respectively.

Hypo-trimethylation of Histone H3 Lysine 4 and Hyper-tri/dimethylation of Histone H3 Lysine 27 as Epigenetic Markers of Poor Prognosis in Patients with Primary Central Nervous System Lymphoma.

PURPOSE: This study aimed to investigate the methylation status of major histone modification sites in primary central nervous system lymphoma (PCNSL) samples and examine their prognostic roles in patients with PCNSL. MATERIALS AND METHODS: Between 2007 and 2020, 87 patients were histopathologically diagnosed with PCNSL. We performed immunohistochemical staining of the formalin-fixed paraffin-embedded samples of PCNSL for major histone modification sites, such as H3K4, H3K9, H3K27, H3K14, and H3K36. After detection of meaningful methylation sites, we examined histone modification enzymes that induce methylation or demethylation at each site using immunohistochemical staining. The meaningful immunoreactivity was validated by western blotting using fresh tissue of PCNSL. RESULTS: More frequent recurrences were found in hypomethylation of H3K4me3 (p=0.004) and hypermethylation of H3K27me2 (p<0.001) and H3K27me3 (p=0.002). These factors were also statistically related to short PFS and overall survival in the univariate and multivariate analyses. Next, histone modification enzymes inducing the demethylation of H3K4 (lysine-specific demethylase-1/2 and Jumonji AT-rich interactive domain [JARID] 1A-D]) and methylation of H3K27 (enhancer of zeste homolog [EZH]-1/2) were immu- nohistochemically stained. Among them, the immunoreactivity of JARID1A inversely associated with the methylation status of H3K4me3 (R2=-1.431), and immunoreactivity of EZH2 was directly associated with the methylation status of H3K27me2 (R2=0.667) and H3K27me3 (R2=0.604). These results were validated by western blotting in fresh PCNSL samples. CONCLUSION: Our study suggests that hypomethylation of H3K4me3 and hypermethylation of H3K27me2 and H3K27me3 could be associated with poor outcomes in patients with PCNSL and that these relationships are modified by JARID1A and EZH2.

Phlogopite-Reinforced Natural Rubber (NR)/Ethylene-Propylene-Diene Monomer Rubber (EPDM) Composites with Aminosilane Compatibilizer.

Rubber compounding with two or more components has been extensively employed to improve various properties. In particular, natural rubber (NR)/ethylene-propylene-diene monomer rubber (EPDM) blends have found use in tire and automotive parts. Diverse fillers have been applied to NR/EPDM blends to enhance their mechanical properties. In this study, a new class of mineral filler, phlogopite, was incorporated into an NR/EPDM blend to examine the mechanical, curing, elastic, and morphological properties of the resulting material. The combination of aminoethylaminopropyltrimethoxysilane (AEAPS) and stearic acid (SA) compatibilized the NR/EPDM/phlogopite composite, further improving various properties. The enhanced properties were compared with those of NR/EPDM/fillers composed of silica or carbon black (CB). Compared with the NR/EPDM/silica composite, the incompatibilized NR/EPDM/phlogopite composite without AEAPS exhibited poorer properties, but NR/EPDM/phlogopite compatibilized by AEAPS and SA showed improved properties. Most properties of the compatibilized NR/EPDM/phlogopite composite were similar to those of the NR/EPDM/CB composite, except for the lower abrasion resistance. The NR/EPDM/phlogopite/AEAPS rubber composite may potentially be used in various applications by replacing expensive fillers, such as CB.

Expression Profile of Circulating MicroRNAs in Dogs With Cardiac Hypertrophy: A Pilot Study.

This study aimed to identify the expression profile of circulating microRNAs in dogs with eccentric or concentric cardiac hypertrophy. A total of 291 microRNAs in serum samples of five dogs with myxomatous mitral valve degeneration (MMVD) and five dogs with pulmonic stenosis (PS) were compared with those of five healthy dogs using microarray analysis. Results of microarray analysis revealed up-regulation of cfa-miR-130b [fold change (FC) = 2.13, p = 0.014), down-regulation of cfa-miR-375 (FC = 1.51, p = 0.014), cfa-miR-425 (FC = 2.56, p = 0.045), cfa-miR-30d (FC = 3.02, p = 0.047), cfa-miR-151 (FC = 1.89, p = 0.023), cfa-miR-19b (FC = 3.01, p = 0.008), and cfa-let-7g (FC = 2.53, p = 0.015) in MMVD group which showed eccentric cardiac hypertrophy, up-regulation of cfa-miR-346 (FC = 2.74, p = 0.032), down-regulation of cfa-miR-505 (FC = 1.56, p = 0.016) in PS group which showed concentric cardiac hypertrophy, and down-regulation of cfa-miR-30c (FC = 3.45, p = 0.013 in MMVD group; FC = 3.31, p = 0.014 in PS group) and cfa-let-7b (FC = 11.42, p = 0.049 in MMVD group; FC = 5.88, p = 0.01 in PS group) in both MMVD and PS groups. In addition, the unsupervised hierarchical clustering of differentially expressed microRNAs in each group resulted in complete separation of healthy dogs from dogs with heart diseases. Therefore, eleven microRNAs among 291 microRNAs were identified as differentially expressed circulating microRNAs related to MMVD or PS in dogs. This pilot study demonstrates that the microRNAs identified in this study could be possible candidates for novel biomarker or therapeutic target related to cardiac hypertrophy in dogs.

Epigenetic Role of Histone Lysine Methyltransferase and Demethylase on the Expression of Transcription Factors Associated with the Epithelial-to-Mesenchymal Transition of Lung Adenocarcinoma Metastasis to the Brain.

PURPOSE: The objective of this study was to investigate the epigenetic role of histone lysine methylation/demethylation on the expression of epithelial-to-mesenchymal transition (EMT) associated transcriptional factors (TFs) during the metastasis of lung adenocarcinoma to the brain. METHODS: Paired samples of lung adenocarcinoma and brain metastasis (BM) were analyzed in 46 individual patients. Both samples were obtained by surgical resection or biopsy of the lung and brain. The paraffin-fixed formalin-embedded samples were obtained from the pathology archives in our institute. In samples of lung adenocarcinoma and BM, immunohistochemical staining was performed for epithelial markers, mesenchymal markers, EMT-TFs, histone lysine methyltransferase and demethylase. RESULTS: The immunoreactivity of EMT-TFs such as Slug (15.6% vs. 42.6%, p = 0.005), Twist (23.6% vs. 45.9%, p = 0.010) and ZEB1 (15.0% vs. 55.9%, p = 0.002) was increased in BM compared with that in lung adenocarcinoma. Epigenetic inducers such as H3K4 methyltransferase (MLL4, p = 0.018) and H3K36me3 demethylase (UTX, p = 0.003) were statistically increased, and epigenetic repressors such as EZH2 (H3K27 methyltransferase, p = 0.046) were significantly decreased in BM compared with those in lung adenocarcinoma. The expression of UTX-ZEB1 (R2 linear = 1.204) and MLL4-Slug (R2 linear = 0.987) was increased in direct proportion, and EZH2-Twist (R2 linear = -2.723) decreased in reverse proportion. CONCLUSIONS: The results suggest that certain histone lysine methyltransferase/demethylase, such as MLL4, UTX, and EZH2, regulate the expression of EMT-TFs such as Slug, ZEB1, and Twist epigenetically, which may thereby influence cancer metastasis from the lung to the brain.

Efficient mutation screening for cervical cancers from circulating tumor DNA in blood.

BACKGROUND: Early diagnosis and continuous monitoring are necessary for an efficient management of cervical cancers (CC). Liquid biopsy, such as detecting circulating tumor DNA (ctDNA) from blood, is a simple, non-invasive method for testing and monitoring cancer markers. However, tumor-specific alterations in ctDNA have not been extensively investigated or compared to other circulating biomarkers in the diagnosis and monitoring of the CC. Therfore, Next-generation sequencing (NGS) analysis with blood samples can be a new approach for highly accurate diagnosis and monitoring of the CC. METHOD: Using a bioinformatics approach, we designed a panel of 24 genes associated with CC to detect and characterize patterns of somatic single-nucleotide variations, indels, and copy number variations. Our NGS CC panel covers most of the genes in The Cancer Genome Atlas (TCGA) as well as additional cancer driver and tumor suppressor genes. We profiled the variants in ctDNA from 24 CC patients who were being treated with systemic chemotherapy and local radiotherapy at the Jeonbuk National University Hospital, Korea. RESULT: Eighteen out of 24 genes in our NGS CC panel had mutations across the 24 CC patients, including somatic alterations of mutated genes (ZFHX3-83%, KMT2C-79%, KMT2D-79%, NSD1-67%, ATM-38% and RNF213-27%). We demonstrated that the RNF213 mutation could be used potentially used as a monitoring marker for response to chemo- and radiotherapy. CONCLUSION: We developed our NGS CC panel and demostrated that our NGS panel can be useful for the diagnosis and monitoring of the CC, since the panel detected the common somatic variations in CC patients and we observed how these genetic variations change according to the treatment pattern of the patient.

Effects of dispersed copper nanoparticles on Ni-ceria based dry methanol fuelled low temperature solid oxide fuel cells.

Methanol is an attractive energy source due to its portability and thermodynamic coke resistance by its oxygen content. In order to operate dry methanol fuel low temperature solid oxide fuel cells (LT-SOFCs), it is important to solve the problems of carbon formation and its low performance. In this study, copper impregnation was selected to decrease the carbon deposition and enhance the performance at low temperature. The interaction of copper, ceria and nickel improves CO oxidation capacity which improves coke tolerance and nano-sized nickel copper alloys improved durability and catalytic performance under methanol feed. It markedly amplified the performance about 0.4 W cm-2 at 550 °C with the durable operation at 1.4 A cm-2 over 50 h. Loading copper nanoparticles is promising method for Ni-ceria based LT-SOFC using methanol fuel with high performance and stable operation.

Osteological Development of the Larvae and Juvenile of Trident Goby, Tridentiger obscurus.

This study is to observe the developmental process of the larval skeleton according to the growth of the trident goby, Tridentiger obscurus belonging to the larvae and juveniles and use it as the basic data of the taxonomic study. 8 days after hatching, the parasphenoid was ossified with an average total length of 3.62 mm, and basioccipital began to ossify. Caudal vertebrae and neural spine ossified in vertebra. 17 days after hatching, the average total length of the long hairs was 4.32 mm, pterotic and epiotic were ossified, and interhyal and subopercle were ossified. 52 days after hatching, the average total length of the juvenile was 18.2 mm, and lateral ethmoid, hypohyal ossified, vertebrae were parapophysis, and epural bone was osseous to the bone.

Osteological Development of the Larvae and Juvenile of Bullhead torrent catfish, Liobagrus obesus.

This study was conducted to investigate the skeletal development of bullhead torrent catfish, Liobagrus obesus larvae and to utilize them as basic data for the taxonomic study of Liobagrus larvae. Skeletal development was observed by being divided into cranium, visceral skeleton, shoulder girdle bone, pelvic girdle bone and vertebra. On the first day after hatching, the pre-larvae had an average total length of 7.92 mm, and a line-shaped parasphenoid ossified in the cranium. In the jaw bone, the dentary supporting the lower jaw and the maxillary supporting the upper jaw were ossified. In the anterior abdominal vertebrae of the vertebra, seven centrums began to ossify and five neural spines ossified simultaneously. On the 3 day after hatching, pre-larvae had an average total length of 8.95 mm, and the prefrontal ossified in cranium. The number of abdominal vertebrae was increased to 14, and three parapophysis developed from the front side. On the 24th day after hatching, post-larvae had an average total length of 15.2 mm and the epural bone ossified in coccyx. The parhypural bone was ossified, and ossification of coccyx and pelvic girdle bone was completed. On the 30th day after hatching, the average total length of the juvenile was 17.8 mm, and the ossification of cranium and visceral skeleton was all completed while the preorbital and three suborbitals were ossified in the orbital region of the cranium.

Protein arginine methyltransferase 7-mediated microRNA-221 repression maintains Oct4, Nanog, and Sox2 levels in mouse embryonic stem cells.

The stemness maintenance of embryonic stem cells (ESCs) requires pluripotency transcription factors, including Oct4, Nanog, and Sox2. We have previously reported that protein arginine methyltransferase 7 (PRMT7), an epigenetic modifier, is an essential pluripotency factor that maintains the stemness of mouse ESCs, at least in part, by down-regulating the expression of the anti-stemness microRNA (miRNA) miR-24-2. To gain greater insight into the molecular basis underlying PRMT7-mediated maintenance of mouse ESC stemness, we searched for new PRMT7-down-regulated anti-stemness miRNAs. Here, we show that miR-221 gene-encoded miR-221-3p and miR-221-5p are anti-stemness miRNAs whose expression levels in mouse ESCs are directly repressed by PRMT7. Notably, both miR-221-3p and miR-221-5p targeted the 3' untranslated regions of mRNA transcripts of the major pluripotency factors Oct4, Nanog, and Sox2 to antagonize mouse ESC stemness. Moreover, miR-221-5p silenced also the expression of its own transcriptional repressor PRMT7. Transfection of miR-221-3p and miR-221-5p mimics induced spontaneous differentiation of mouse ESCs. CRISPR-mediated deletion of the miR-221 gene, as well as specific antisense inhibitors of miR-221-3p and miR-221-5p, inhibited the spontaneous differentiation of PRMT7-depleted mouse ESCs. Taken together, these findings reveal that the PRMT7-mediated repression of miR-221-3p and miR-221-5p expression plays a critical role in maintaining mouse ESC stemness. Our results also establish miR-221-3p and miR-221-5p as anti-stemness miRNAs that target Oct4, Nanog, and Sox2 mRNAs in mouse ESCs.

Parturition and Morphological Development of Larvae and Juvenile in Hippocampus kuda from Korea.

The newborn, Hippocampus kuda larvae were 6.97-8.81, standard length (SL) mm (mean 7.89 mm) and mouth and anuse were open. Dorsal fin rays 15-18 and pectoral fin-rays were counted 8 and had 10-11+21 myotomes, body's bony plate ring being developed strongly in the central axis of myotomes part. 4 days after bearing, the SL was 7.02-9.47 mm (mean 8.24 mm) and nostrils began to open. 12 days after bearing, larvae attained to 8.91-11.2 SL mm (mean 10.0 mm). From this time, their unique predation habit appeared. 21 days bearing, larvae attained to 12.1-14.8 SL mm (mean 13.4 mm) the and thorn of back was enlarged among the plate formed around ring. 41 days bearing, seahorses attained to 17.1-17.8 SL mm (mean 17.4 mm) and the number of body's bony plate ring of the top of rings trunk was 11 and on the tail of them was 33-36, similar to figure of adult.

Increased expression of the histone H3 lysine 4 methyltransferase MLL4 and the histone H3 lysine 27 demethylase UTX prolonging the overall survival of patients with glioblastoma and a methylated MGMT promoter.

OBJECTIVE The purpose of the present study was to investigate the epigenetic and prognostic roles of an H3K4 methyltransferase (mixed lineage leukemia 4 [MLL4]) and H3K27 demethylase (ubiquitously transcribed tetratricopeptide repeat gene on X chromosome [UTX]) in progression-free survival (PFS) and overall survival (OS) of patients with glioblastoma (GBM) who were treated with radiotherapy, chemotherapy, or both after resection. In addition, the authors examined methylation at the promoter of the O-6-methylguanine-DNA methyltransferase ( MGMT) gene and other prognostic factors predicting length of PFS and OS in these patients. METHODS The medical records of 76 patients having a new diagnosis of histologically ascertained GBM in the period of January 2002 to December 2013 at the authors' institution were retrospectively reviewed. Immunohistochemical staining for MLL4 and UTX was performed on archived paraffin-embedded tissues obtained by biopsy or resection. The methylation status of the MGMT promoter in these tissues was determined by methylation-specific PCR analysis. RESULTS During the follow-up period (mean length 18.1 months, range 4.1-43.5 months), 68 (89.5%) of the patients died. The MGMT promoter was methylated in 49 patients (64.5%) and unmethylated in 27 (35.5%). The immunoreactivity pattern of UTX was identical to that of MLL4; increased expression of these 2 proteins was observed in samples from 34 patients (44.7%) and decreased expression in 42 patients (55.3%). The mean length of PFS was 9.2 months (95% CI 6.8-11.6 months). Extent of surgery, recursive partitioning analysis (RPA) class, and methylation status of the MGMT promoter were all associated with increased PFS in the multivariate analysis of factors predicting PFS. The mean length of OS was 18.6 months (95% CI 14.3-22.9 months). Patient age (p = 0.004), WHO performance status score (p = 0.019), extent of surgery (p = 0.007), RPA class (p = 0.036), methylation status of the MGMT promoter (p = 0.010), and increased expression of UTX-MLL4 (p = 0.001) were significantly associated with increased OS in multivariate analysis. Interestingly, in patients with an unmethylated MGMT promoter, immunoreactivity of UTX-MLL4 was not associated with changes in OS (p = 0.350). However, in the patients with a methylated MGMT promoter, increased UTX-MLL4 expression was strongly associated with increased OS (p < 0.001). CONCLUSIONS The results of this study suggest that increased expression of UTX-MLL4 positively influences the outcome of patients with GBM having a methylated MGMT promoter. Therefore, UTX-MLL4 immunoreactivity could be a useful predictor of the response to conventional treatment with radiotherapy or chemotherapy among GBM patients whose tumors have a methylated MGMT promoter.

A feedback loop comprising PRMT7 and miR-24-2 interplays with Oct4, Nanog, Klf4 and c-Myc to regulate stemness.

Self-renewal and pluripotency are two fundamental characteristics of embryonic stem cells (ESCs) and are controlled by diverse regulatory factors, including pluripotent factors, epigenetic regulators and microRNAs (miRNAs). Although histone methyltransferases are key epigenetic regulators, whether and how a histone methyltransferase forms a network with miRNAs and the core pluripotent factor system to regulate ESC stemness is little known. Here, we show that the protein arginine methyltransferase 7 (PRMT7) is a pluripotent factor essential for the stemness of mouse ESCs. PRMT7 repressed the miR-24-2 gene encoding miR-24-3p and miR-24-2-5p by upregulating the levels of symmetrically dimethylated H4R3. Notably, miR-24-3p targeted the 3' untranslated regions (UTRs) of the major pluripotent factors Oct4, Nanog, Klf4 and c-Myc, whereas miR-24-2-5p silenced Klf4 and c-Myc expression. miR-24-3p and miR-24-2-5p also targeted the 3'UTR of their repressor gene Prmt7 miR-24-3p and miR-24-2-5p induced mouse ESC differentiation, and their anti-sense inhibitors substantially reversed spontaneous differentiation of PRMT7-depleted mouse ESCs. Oct4, Nanog, Klf4 and c-Myc positively regulated Prmt7 expression. These findings define miR-24-3p and miR-24-2-5p as new anti-pluripotent miRNAs and also reveal a novel epigenetic stemness-regulatory mechanism in which a double-negative feedback loop consisting of PRMT7 and miR-24-3p/miR24-2-5p interplays with Oct4, Nanog, Klf4 and c-Myc to control ESC stemness.

The application of catalyst-recovered SnO2 as an anode material for lithium secondary batteries.

We studied the electrochemical characteristics of tin dioxide (SnO2) recovered from waste catalyst material which had been previously used in a polymer synthesis reaction. In order to improve the electrochemical performance of the SnO2 anode electrode, we synthesized a nanocomposite of recovered SnO2 and commercial iron oxide (Fe2O3) (weight ratio 95:5) using a solid state method. X-ray diffraction (XRD) and field emission scanning electron microscopy (FE-SEM) analyses revealed an additional iron oxide phase within a porous nanocomposite architecture. The electrochemical characterizations were based on galvanostatic charge-discharge (CD) curves, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). In the first discharge, the capacity of the SnO2-Fe2O3 nanocomposite was 1700 mAh g(-1), but was reduced to about 1200 mAh g(-1) in the second discharge. Thereafter, a discharge capacity of about 1000 mAh g(-1)was maintained up to the 20th cycle. The SnO2-Fe2O3 nanocomposite showed better reversible capacities and rate capabilities than either the recovered SnO2 or commercial Fe2O3 nanoparticle samples.

An essential role for UTX in resolution and activation of bivalent promoters.

Trimethylated histone H3 lysine 27 (H3K27me3) is linked to gene silencing, whereas H3K4me3 is associated with gene activation. These two marks frequently co-occupy gene promoters, forming bivalent domains. Bivalency signifies repressed but activatable states of gene expression and can be resolved to active, H3K4me3-prevalent states during multiple cellular processes, including differentiation, development and epithelial mesenchymal transition. However, the molecular mechanism underlying bivalency resolution remains largely unknown. Here, we show that the H3K27 demethylase UTX (also called KDM6A) is required for the resolution and activation of numerous retinoic acid (RA)-inducible bivalent genes during the RA-driven differentiation of mouse embryonic stem cells (ESCs). Notably, UTX loss in mouse ESCs inhibited the RA-driven bivalency resolution and activation of most developmentally critical homeobox (Hox) a-d genes. The UTX-mediated resolution and activation of many bivalent Hox genes during mouse ESC differentiation were recapitulated during RA-driven differentiation of human NT2/D1 embryonal carcinoma cells. In support of the importance of UTX in bivalency resolution, Utx-null mouse ESCs and UTX-depleted NT2/D1 cells displayed defects in RA-driven cellular differentiation. Our results define UTX as a bivalency-resolving histone modifier necessary for stem cell differentiation.