[A Case of Consciousness Disorder Induced by the Syndrome of Inappropriate Antidiuretic Hormone Secretion Following Cisplatin and 5-Fluorouracil Chemotherapy in a Patient with Tongue Cancer].

We herein report a case of a consciousness disorder that was induced by the syndrome of inappropriate antidiuretic hormone secretion following cisplatin (CDDP) and 5 -fluorouracil (5-FU) chemotherapy in a patient with tongue cancer. A 72- year-old woman complained of tongue pain and was admitted to our hospital for neoadjuvant chemotherapy, under a diagnosis of tongue squamous cell carcinoma (T4aN2bM0). She was treated with CDDP and 5-FU. On the second day after administration, she complained of nausea and anorexia, and on the third day, she showed impaired consciousness. Laboratory studies revealed that the patient had a serum sodium concentration 112mEq/L, and no dehydration was noted. The patient was diagnosed with SIADH, using the appropriate diagnostic criteria based on serum and urine hypoosmolality. We subsequently discontinued chemotherapy and initiated fluid restriction and sodium supplements. Two days after this treatment, her consciousness level improved, and on the fifth day of treatment, laboratory studies revealed a serum sodium level of 134mEq/ L.

Efficient screening of long terminal repeat retrotransposons that show high insertion polymorphism via high-throughput sequencing of the primer binding site.

Retrotransposons have been used frequently for the development of molecular markers by using their insertion polymorphisms among cultivars, because multiple copies of these elements are dispersed throughout the genome and inserted copies are inherited genetically. Although a large number of long terminal repeat (LTR) retrotransposon families exist in the higher eukaryotic genomes, the identification of families that show high insertion polymorphism has been challenging. Here, we performed an efficient screening of these retrotransposon families using an Illumina HiSeq2000 sequencing platform with comprehensive LTR library construction based on the primer binding site (PBS), which is located adjacent to the 5' LTR and has a motif that is universal and conserved among LTR retrotransposon families. The paired-end sequencing library of the fragments containing a large number of LTR sequences and their insertion sites was sequenced for seven strawberry (Fragaria × ananassa Duchesne) cultivars and one diploid wild species (Fragaria vesca L.). Among them, we screened 24 families with a "unique" insertion site that appeared only in one cultivar and not in any others, assuming that this type of insertion should have occurred quite recently. Finally, we confirmed experimentally the selected LTR families showed high insertion polymorphisms among closely related cultivars.

Hazards associated with blue light emitted during gas metal arc welding of mild steel using various shielding gases and currents.

Blue light emitted during arc welding is known to potentially cause photoretinopathy. To help prevent retinal injury, it is important to identify the hazards associated with various welding conditions. The present work conducted experiments involving gas metal arc welding of mild steel under various conditions, and measured the spectral radiance of the arcs. The effective radiance values, as used by the American Conference of Governmental Industrial Hygienists (ACGIH) to quantify exposure level of blue light, were subsequently calculated from the data. The resulting values were in the range of 5.0-118 W/cm2/sr, corresponding to hazardous levels according to ACGIH guidelines. The effective radiance was increased at higher welding currents and when pulsed currents were used rather than steady currents. The blue light hazard was also affected by the type of shielding gas employed. These data confirm that it is very hazardous to stare at arcs during gas metal arc welding of mild steel. As such, appropriate eye protection is necessary during arc welding, and directly staring at the arc should be avoided.

Contribution of dipeptidyl peptidase IV to the severity of dextran sulfate sodium-induced colitis in the early phase.

Dipeptidyl peptidase IV (DPPIV) degrades some peptide hormones and cytokines, resulting in homeostatic modulation. However, the role of DPPIV in inflammatory bowel diseases remains controversial. To determine the role of DPPIV in colitis, we used F344/DuCrlCrlj (F344/Du) rats as a DPPIV-deficient model. The serum DPPIV activity was much lower in the F344/Du rats than in F344/Jcl rats which were used as a DPPIV-positive model. Interestingly, the disease activity index (DAI) was different in the early phase of 2% dextran sulfate sodium (DSS)-induced colitis, as judged by the mucosal myeloperoxidase activity, colonic weight, and cecal fermentation. Similarly, retarded DAI was apparent in the DPPIV-deficient rats with 1% DSS-induced colitis. These findings suggest that a low level of DPPIV activity contributed to maintaining intestinal homeostasis by suppressing the cleavage of cytokines and growth hormones in DSS-induced colitis, especially in the early phase of colitis and with moderate inflammation.

bex-db: Bioinformatics workbench for comprehensive analysis of barley-expressed genes.

Barley (Hordeum vulgare) is one of the world's most important cereal crops. Although its large and complex genome has held back barley genomics for quite a while, the whole genome sequence was released in 2012 by the International Barley Genome Sequencing Consortium (IBSC). Moreover, more than 30,000 barley full-length cDNAs (FLcDNAs) are now available in the public domain. Here we present the Barley Gene Expression Database (bex-db: http://barleyflc.dna.affrc.go.jp/bexdb/index.html) as a repository of transcriptome data including the sequences and the expression profiles of barley genes resulting from microarray analysis. In addition to FLcDNA sequences, bex-db also contains partial sequences of more than 309,000 novel expressed sequence tags (ESTs). Users can browse the data via keyword, sequence homology and expression profile search options. A genome browser was also developed to display the chromosomal locations of barley FLcDNAs and wheat (Triticum aestivum) transcripts as well as Aegilops tauschii gene models on the IBSC genome sequence for future comparative analysis of orthologs among Triticeae species. The bex-db should provide a useful resource for further genomics studies and development of genome-based tools to enhance the progress of the genetic improvement of cereal crops.

Clinical Characteristics of the Cleft Lip and/or Palate: Association with Congenital Anomalies, Syndromes, and Chromosomal Anomalies.

Background: Cleft lip and/or palate (CL/P) can be accompanied by other congenital anomalies. We conducted a long-term evaluation of the associations between cleft patterns, sex distribution, and accompanying congenital anomalies of patients with CL/P. Methods: The medical records of 739 patients with CL/P, seen between January 1967 and December 2020, were retrospectively reviewed. Fisher's exact test was used for statistical analysis. Results: Among the 739 patients with CL/P, the male-to-female ratio was 1.1. Regarding the cleft pattern, 121 (16.4%), 104 (14.1%), 280 (37.9%), 198 (26.8%), and 36 (4.9%) patients had cleft lip (CL), cleft lip and alveolus (CLA), cleft lip and palate (CLP), cleft palate (CP), and submucous cleft palate (SMCP), respectively. Congenital anomalies were identified in 107 (14.5%) cases, of which 53 (49.5%) had congenital heart disease. The frequencies of congenital anomalies patients with in CL/P were 14/225 (6.2%), 36/280 (12.9%), 43/198 (21.7%), and 14/36 (38.9%) for a combination of CL and CLA, CLP, CP, and SMCP, respectively. Accompanying syndromes and chromosomal anomalies were identified in 40 (5.4%) cases, in which Pierre Robin sequence (16 cases of CP and 4 cases of SMCP) was the most frequent. Conclusion: No sex differences were observed in CL/P, and CLP and CP were the most common cleft patterns. Congenital anomalies associated with CL/P were dominated by congenital heart disease and were most frequently identified in CP and SMCP cases. Notably, the Pierre Robin sequence, a complex syndrome characterized by micrognathia, glossoptosis, respiratory obstruction, and a U- or V-shaped CP, was found in cases of both CP and SMCP, and accounted for the symptoms in most cases.

Bile acid is a host factor that regulates the composition of the cecal microbiota in rats.

BACKGROUND & AIMS: Alterations in the gastrointestinal microbiota have been associated with metabolic diseases. However, little is known about host factors that induce changes in gastrointestinal bacterial populations. We investigated the role of bile acids in this process because of their strong antimicrobial activities, specifically the effects of cholic acid administration on the composition of the gut microbiota in a rat model. METHODS: Rats were fed diets supplemented with different concentrations of cholic acid for 10 days. We used 16S ribosomal RNA gene clone library sequencing and fluorescence in situ hybridization to characterize the composition of the cecal microbiota of the different diet groups. Bile acids in feces, organic acids in cecal contents, and some blood parameters were also analyzed. RESULTS: Administration of cholic acid induced phylum-level alterations in the composition of the gut microbiota; Firmicutes predominated at the expense of Bacteroidetes. Cholic acid feeding simplified the composition of the microbiota, with outgrowth of several bacteria in the classes Clostridia and Erysipelotrichi. Externally administered cholic acid was efficiently transformed into deoxycholic acid by a bacterial 7α-dehydroxylation reaction. Serum levels of adiponectin decreased significantly in rats given the cholic acid diet. CONCLUSIONS: Cholic acid regulates the composition of gut microbiota in rats, inducing similar changes to those induced by high-fat diets. These findings improve our understanding of the relationship between metabolic diseases and the composition of the gastrointestinal microbiota.

Comprehensive sequence analysis of 24,783 barley full-length cDNAs derived from 12 clone libraries.

Full-length cDNA (FLcDNA) libraries consisting of 172,000 clones were constructed from a two-row malting barley cultivar (Hordeum vulgare 'Haruna Nijo') under normal and stressed conditions. After sequencing the clones from both ends and clustering the sequences, a total of 24,783 complete sequences were produced. By removing duplicates between these and publicly available sequences, 22,651 representative sequences were obtained: 17,773 were novel barley FLcDNAs, and 1,699 were barley specific. Highly conserved genes were found in the barley FLcDNA sequences for 721 of 881 rice (Oryza sativa) trait genes with 50% or greater identity. These FLcDNA resources from our Haruna Nijo cDNA libraries and the full-length sequences of representative clones will improve our understanding of the biological functions of genes in barley, which is the cereal crop with the fourth highest production in the world, and will provide a powerful tool for annotating the barley genome sequences that will become available in the near future.

Cancer-associated fibroblasts and CD163-positive macrophages in oral squamous cell carcinoma: their clinicopathological and prognostic significance.

BACKGROUND: Stromal cells are believed to affect cancer invasion and metastasis. The purpose of this study was to evaluate the distribution of cancer-associated fibroblasts (CAFs) and the incidence of tumor-associated macrophages (TAMs) in oral squamous cell carcinoma (OSCC), focusing on clinicopathological factors and patient prognosis, as well as cancer invasion. METHODS: The study included 108 patients with OSCC. Anti-α-smooth muscle actin, CD68, and CD163 antibodies were used to identify CAFs and TAMs. CAFs were divided into 4 grades on the basis of staining intensity: negative (0), scanty (1), focal (2), and abundant (3). The most intensive areas of macrophage concentration in each tumor invasive stroma were also evaluated. RESULTS: The cancer specimens were divided into Grade 0/1, Grade 2, and Grade 3 on the basis of CAF grade. In addition, they were divided into low- and high-grade groups on the basis of the number of CD68-positive and CD163-positive macrophages. The latter were significantly increased in the Grade 2 CAF group compared to the Grade 0/1 group (P = 0.009). Kaplan-Meier and multivariate survival analyses revealed that Grade 2 CAFs (P = 0.003) and high CD163-positive macrophage levels (P = 0.007) significantly correlated with a poor outcome in patients with OSCC, and that a high CD163-positive macrophage level was a significant and an independent prognostic factor (P = 0.045). CONCLUSIONS: Cancer-associated fibroblasts and CD163-positive macrophages may be potential prognostic predictors of OSCC.

12α-Hydroxylated bile acid enhances accumulation of adiponectin and immunoglobulin A in the rat ileum.

We previously reported that dietary supplementation with cholic acid (CA), the primary 12α-hydroxylated (12αOH) bile acid (BA), reduces plasma adiponectin concentration in rats. The aim of this study was to examine the distribution of adiponectin in the body of CA-fed rats and its influence on mucosal immunoglobulin A concentration in the intestine. Rats were fed a diet supplemented with or without CA (0.5 g CA/kg diet) for 13 weeks. A reduction in plasma adiponectin level was observed from week 3. At the end of the experiment, the CA diet reduced plasma adiponectin concentration both in the portal and aortic plasma. Accumulation of adiponectin was accompanied by an increase in cadherin-13 mRNA expression in the ileal mucosa of CA-fed rats. No increase was observed in adiponectin mRNA expression in the ileal and adipose tissues of the CA-fed rats. Immunoglobulin A concentration in the ileal mucosa was elevated in the CA-fed rats and was correlated with the ileal adiponectin concentration. 12αOH BAs may modulate mucosal immune response that are involved in the accumulation of adiponectin in the ileum.

Fume particle size distribution and fume generation rate during arc welding of cast iron.

This study measured the fume particle size distribution and fume generation rate during arc welding of cast iron and estimated the generation rate of respirable dust. In addition, the generation rate of particles with a diameter of 0.3 µm or less was estimated. In this experiment, three types of filler materials (mild steel wire, stainless steel wire, and mild steel covered electrodes) with main constituents of Fe or Fe-Cr-Ni, as proposed previously by the authors, were used. The welding methods were gas metal arc welding and shielded metal arc welding. The fumes measured in this research contained 73-91% respirable dust, and the fume generation rates were in the range of 1.96-12.2 mg/s. The results of this study were as follows: (i) the welding current affects the generation rate of respirable dust, and it is highly likely that the higher the fume generation rate, the more respirable dust is generated; (ii) the generation rates of respirable dust at low and high current were highest when mild steel covered electrodes and stainless steel wire was used, respectively; and (iii) the generation rate of particles with a diameter of 0.3 µm or less was highest when stainless steel wire is used.

Comprehensive analysis of hazard of ultraviolet radiation emitted during arc welding of cast iron.

OBJECTIVES: Ultraviolet radiation (UVR) emitted during arc welding frequently causes keratoconjunctivitis and skin erythema. The extent of the hazard of UVR varies depending on the welding process and conditions. Therefore, it is important to identify the levels of UVR present under different conditions. The purpose of this study was to investigate the degree of hazard of UVR emitted by the different types of arc welding of cast iron frequently used in industry. METHODS: In this study, we experimentally measured the UVR emitted during gas tungsten arc welding (GTAW), shielded metal arc welding (SMAW), and gas metal arc welding (GMAW) of cast iron. The degree of hazard of UVR was quantitatively evaluated in accordance with the guidelines of the American Conference of Governmental Industrial Hygienists. RESULTS: Effective irradiances measured in this study were in the range 0.045-2.2 mW/cm2 at a distance of 500 mm from the welding arc. The maximum allowable exposure times corresponding to these levels were only 1.4-67 s/day. CONCLUSIONS: UVR emitted during arc welding of cast iron has the following characteristics: (a) It is more hazardous at higher welding currents. (b) The magnitude of the hazard, which depends on the welding process, increases in the order of GMAW > SMAW > GTAW. (c) It is influenced by the filler material used; that is, the components contained in the filler material affect the hazard of UVR. The effect is Fe > Ni, Cr.

Clinical Features of Oral Multiple Primary Carcinomas Compared with Oral Single Primary Carcinoma.

BACKGROUND: Owing to the increase in the older population and the increased life span, the number of patients with oral multiple primary carcinomas will increase. Predicting the second and third carcinoma clinically is difficult, and the presence of second or third carcinomas is a factor that determines the prognosis of oral carcinoma. In this study, we examined the clinical features of oral multiple primary carcinomas treated in our department. METHODS: We retrospectively reviewed the medical records of patients with oral squamous cell carcinoma who underwent radical treatment at and were followed by the Department of Oral and Maxillofacial Surgery, Tottori University Hospital from January 2003 to October 2017. RESULTS: This study included 261 patients: 241 patients had oral single primary carcinoma and 20 patients had oral multiple primary carcinomas. Oral multiple primary carcinomas showed female predilection and occurred more frequently in the lower gingiva and significantly less frequently in the tongue (P < 0.01). Oral multiple primary carcinomas showed a significantly higher recurrence rate (P < 0.01). The 5-year overall survival of oral single primary carcinoma patients was 88.0% compared with 95% for oral multiple primary carcinomas, with no significant difference (log rank test, P = 0.54). However, the 15-year survival rate dropped to 28.1% in oral multiple primary carcinomas. The cumulative disease incidence rates of metachronous second primary carcinoma from first carcinoma at 5 years and 10 years were 3.45% and 5.36%, respectively. CONCLUSION: Oral multiple primary carcinomas rarely occur in the tongue. The 5-year survival rate showed no difference between single and multiple carcinoma patients, but over longer observation, the prognosis of multiple carcinoma was poor owing to a high recurrence rate. Because of the high recurrence rate and risk of further metachronous carcinoma in oral multiple primary carcinomas, longer-term follow-up is required.

Study on the Development of a New Device with Dual Cameras for Evaluating Expiratory Nasal Flow.

BACKGROUND: Use of the Glatzel mirror for measuring expiratory nasal flow in preschool children has the disadvantage of vagueness, and the mirror may induce fear and inhibition of interest in those children. In response to these limitations, we developed a new device with dual cameras for measuring expiratory nasal flow in 2 to 6 year old children. The aim of this study is to compare the Glatzel mirror and the new device, in terms of accurate assessment of expiratory nasal flow, children's feelings, and correlation to each child's profile. METHODS: This study evaluated 20 cleft lip and palate patients and 21 healthy children aged between 2 and 6 (under 7) years. After consent was granted, a 4-week screening period was undertaken followed by inspection at weeks 8, 16, 24, and 32. Each inspection was conducted while the children were asked to pronounce various sounds and comprised three stages: i) use of the Glatzel mirror, ii) subjective visual assessment using the new device, and iii) image recording by dual cameras of the new device. Questionnaires for the new device were administered at the initial and final inspections. To contrast the results between the Glatzel mirror and the new device, the numbers that indicated values of subjective visual assessment and camera assessment greater than the assessment values of the Glatzel mirror were compared. For measuring the children's responses to the new device compared with those to the Glatzel mirror, the answers to the questionnaires were compared. For the comparison of the children's profiles (age and sex) and feelings, the numbers of subjects who could use the new device were measured. RESULTS: The camera assessment of the new device indicated significantly greater values than that of the Glatzel mirror (P < 0.05). The feelings of the subjects to the new device mostly improved as the study progressed. Subjects aged 3 years and older were generally able to use the new device from the initial inspection. For both sexes, as the inspection progressed, the number occasions of successful use increased. CONCLUSION: This study demonstrated the superiority of the new device with dual cameras to the Glatzel mirror in terms of functionality and attitude of children.

The Scaly-foot Snail genome and implications for the origins of biomineralised armour.

The Scaly-foot Snail, Chrysomallon squamiferum, presents a combination of biomineralised features, reminiscent of enigmatic early fossil taxa with complex shells and sclerites such as sachtids, but in a recently-diverged living species which even has iron-infused hard parts. Thus the Scaly-foot Snail is an ideal model to study the genomic mechanisms underlying the evolutionary diversification of biomineralised armour. Here, we present a high-quality whole-genome assembly and tissue-specific transcriptomic data, and show that scale and shell formation in the Scaly-foot Snail employ independent subsets of 25 highly-expressed transcription factors. Comparisons with other lophotrochozoan genomes imply that this biomineralisation toolkit is ancient, though expression patterns differ across major lineages. We suggest that the ability of lophotrochozoan lineages to generate a wide range of hard parts, exemplified by the remarkable morphological disparity in Mollusca, draws on a capacity for dynamic modification of the expression and positioning of toolkit elements across the genome.

Alteration of Bile Acid Metabolism by a High-Fat Diet Is Associated with Plasma Transaminase Activities and Glucose Intolerance in Rats.

Ingestion of a high-fat (HF) diet is known to enhance bile acid (BA) secretion, but precise information about the BA molecular species is lacking, especially information on the conjugated BAs in enterohepatic circulation. As cholesterol is the precursor of BAs, we analyzed alterations of the entire BA metabolic pathway in response to a HF diet without the addition of cholesterol and BA in the diet. Additionally, we evaluated the relationships between BA metabolism and some disorders, such as plasma transaminase activities and glucose intolerance induced by the HF diet. Acclimated WKAH/HkmSlc male rats (3 wk old) were divided into two groups fed a control or the HF diet for 22 wk. Fasting blood glucose was measured during the experimental period, and an intraperitoneal glucose tolerance test was performed at week 21. As a result, ingestion of the HF diet selectively increased the concentration of taurocholic acid in the bile and small intestinal contents as well as deoxycholic acid in the large intestinal contents and feces. These results indicated a selective increase of 12α-hydroxylated BA concentrations in response to the HF diet. Moreover, fecal 12α-hydroxylated BA concentration was positively correlated with cumulative energy intake, visceral adipose tissue weight, and glucose intolerance. The present study suggests that fecal 12α-hydroxylated BA is a non-invasive marker that can detect the early phase of glucose intolerance.

Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.

The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKK(250) On SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.

The entire organization of transcription units on the Bacillus subtilis genome.

BACKGROUND: In the post-genomic era, comprehension of cellular processes and systems requires global and non-targeted approaches to handle vast amounts of biological information. RESULTS: The present study predicts transcription units (TUs) in Bacillus subtilis, based on an integrated approach involving DNA sequence and transcriptome analyses. First, co-expressed gene clusters are predicted by calculating the Pearson correlation coefficients of adjacent genes for all the genes in a series that are transcribed in the same direction with no intervening gene transcribed in the opposite direction. Transcription factor (TF) binding sites are then predicted by detecting statistically significant TF binding sequences on the genome using a position weight matrix. This matrix is a convenient way to identify sites that are more highly conserved than others in the entire genome because any sequence that differs from a consensus sequence has a lower score. We identify genes regulated by each of the TFs by comparing gene expression between wild-type and TF mutants using a one-sided test. By applying the integrated approach to 11 sigma factors and 17 TFs of B. subtilis, we are able to identify fewer candidates for genes regulated by the TFs than were identified using any single approach, and also detect the known TUs efficiently. CONCLUSION: This integrated approach is, therefore, an efficient tool for narrowing searches for candidate genes regulated by TFs, identifying TUs, and estimating roles of the sigma factors and TFs in cellular processes and functions of genes composing the TUs.

Blue-Light Hazard From Gas Metal Arc Welding of Aluminum Alloys.

OBJECTIVES: The objective was to quantify the blue-light hazard from gas metal arc welding (GMAW) of aluminum alloys. The exposure level is expected to depend on the welding conditions. Therefore, it is important to identify the blue-light hazard under various welding conditions. METHODS: We experimentally conducted GMAW of aluminum alloys under various welding conditions and measured the spectral radiance of the arcs. The effective blue-light radiance, which the American Conference of Governmental Industrial Hygienists has defined to quantify the exposure level of blue light, was calculated from the measured spectral radiance. The maximum acceptable exposure duration per 10000 s for this effective blue-light radiance was calculated. RESULTS: The effective blue-light radiance measured in this study was in the range of 2.9-20.0 W cm-2·sr. The corresponding maximum acceptable exposure duration per 10000 s was only 5.0-34 s, so it is hazardous to view the welding arc. The effective blue-light radiance was higher at higher welding currents than at lower welding currents, when pulsed welding currents were used rather than steady welding currents, and when magnesium was included in the welding materials. CONCLUSIONS: It is very hazardous to view the arcs in GMAW of aluminum alloys. Welders and their helpers should use appropriate eye protection in arc-welding operations. They should also avoid direct light exposure when starting an arc-welding operation.

DNA Methylome Analysis Identifies Transcription Factor-Based Epigenomic Signatures of Multilineage Competence in Neural Stem/Progenitor Cells.

Regulation of the epigenome during in vivo specification of brain stem cells is still poorly understood. Here, we report DNA methylome analyses of directly sampled cortical neural stem and progenitor cells (NS/PCs) at different development stages, as well as those of terminally differentiated cortical neurons, astrocytes, and oligodendrocytes. We found that sequential specification of cortical NS/PCs is regulated by two successive waves of demethylation at early and late development stages, which are responsible for the establishment of neuron- and glia-specific low-methylated regions (LMRs), respectively. The regulatory role of demethylation of the gliogenic genes was substantiated by the enrichment of nuclear factor I (NFI)-binding sites. We provide evidence that de novo DNA methylation of neuron-specific LMRs establishes glia-specific epigenotypes, essentially by silencing neuronal genes. Our data highlight the in vivo implications of DNA methylation dynamics in shaping epigenomic features that confer the differentiation potential of NS/PCs sequentially during development.