Premature termination of reprogramming in vivo leads to cancer development through altered epigenetic regulation.

Cancer is believed to arise primarily through accumulation of genetic mutations. Although induced pluripotent stem cell (iPSC) generation does not require changes in genomic sequence, iPSCs acquire unlimited growth potential, a characteristic shared with cancer cells. Here, we describe a murine system in which reprogramming factor expression in vivo can be controlled temporally with doxycycline (Dox). Notably, transient expression of reprogramming factors in vivo results in tumor development in various tissues consisting of undifferentiated dysplastic cells exhibiting global changes in DNA methylation patterns. The Dox-withdrawn tumors arising in the kidney share a number of characteristics with Wilms tumor, a common pediatric kidney cancer. We also demonstrate that iPSCs derived from Dox-withdrawn kidney tumor cells give rise to nonneoplastic kidney cells in mice, proving that they have not undergone irreversible genetic transformation. These findings suggest that epigenetic regulation associated with iPSC derivation may drive development of particular types of cancer.

[Calcified Amorphous Tumor Diagnosed After Stroke:Report of a Case].

Calcified amorphous tumor (CAT), a non-neoplastic tumor, is rare. Histopathologic features are the presence of calcified nodules in an amorphous background of fibrin. CAT is reported to be associated with renal dysfunction or hemodialysis, and possibly causes cerebral embolism. We report a case of CAT diagnosed after stroke. A 58-year-old male with a 2-year history of hemodialysis was diagnosed with an acute stroke, and was treated medically. Paralysis promptly improved, but transthoracic echocardiography revealed a tumor attached to the posterior mitral leaflet and dense mitral annular calcification. To prevent embolism due to the large tumor, we performed resection of the tumor. Pathological findings showed calcifications surrounded by amorphous fibrous tissue, indicating CAT. Postoperative course was uneventful.

PubChemQC B3LYP/6-31G*//PM6 Data Set: The Electronic Structures of 86 Million Molecules Using B3LYP/6-31G* Calculations.

The presented "PubChemQC B3LYP/6-31G*//PM6" data set is composed of the electronic properties of 85,938,443 molecules, encompassing a broad spectrum of molecules from essential compounds to biomolecules with a molecular weight up to 1000. These molecules account for 94.0% of the original PubChem Compound catalog as of August 29, 2016. The electronic properties, including orbitals, orbital energies, total energies, dipole moments, and other pertinent properties, were computed by using the B3LYP/6-31G* and PM6 methods. The data set, available in three formats, namely, GAMESS quantum chemistry program files, selected JSON output files, and a PostgreSQL database, provides researchers with the ability to query molecular properties. It is further subdivided into five subdata sets for more specific data. The first two subsets encompass molecules with carbon, hydrogen, oxygen, and nitrogen with molecular weights under 300 and 500, respectively. The third and fourth subsets incorporate molecules with carbon, hydrogen, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, with molecular weights under 300 and 500, respectively. The fifth subset comprises molecules with carbon, hydrogen, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, sodium, potassium, magnesium, and calcium, with a molecular weight of under 500. The coefficients of determination for the highest occupied molecular orbital-lowest unoccupied molecular orbital energy gap range from 0.892 (for CHON500) to 0.803 (for the whole data set). These comprehensive results pave the way for applications in drug discovery and materials science, among others. The data sets can be accessed under the Creative Commons Attribution 4.0 International license at the following web address: https://nakatamaho.riken.jp/pubchemqc.riken.jp/b3lyp_pm6_datasets.html.

Comparison of Phenotypes in Subcutaneous Fat and Perivascular Adipose Tissue Surrounding the Saphenous Vein in Coronary Artery Bypass Grafting.

BACKGROUND: The saphenous vein (SV) is used as an essential conduit in coronary artery bypass grafting (CABG), but the long-term patency of SV grafts is a crucial issue. The use of the novel "no-touch" technique of harvesting the SV together with its surrounding tissue has been reported to result in good long-term graft patency of SV grafts. We recently showed that perivascular adipose tissue (PVAT) surrounding the SV (SV-PVAT) had lower levels of metaflammation and consecutive adipose tissue remodeling than did PVAT surrounding the coronary artery. However, the difference between SV-PVAT and subcutaneous adipose tissue (SCAT) remains unclear.Methods and Results: Fat pads were sampled from 55 patients (38 men, 17 women; mean [±SD] age 71±8 years) with coronary artery disease who underwent elective CABG. Adipocyte size was significantly larger in SV-PVAT than SCAT. The extent of fibrosis was smaller in SV-PVAT than SCAT. There were no significant differences between SCAT and SV-PVAT in macrophage infiltration area, quantified by antibodies for CD68, CD11c, and CD206, or in gene expression levels of metaflammation-related markers. Expression patterns of adipocyte developmental and pattern-forming genes differed between SCAT and SV-PVAT. CONCLUSIONS: The properties of SV-PVAT are close to, but not the same as, those of SCAT, possibly resulting from inherent differences in adipocytes. SV-PVAT has healthy expansion with less fibrosis in fat than SCAT.

[Acute Coronary Syndrome in a Patient with Giant Coronary Artery Aneurysm:Report of a Case].

Coronary artery aneurysm is a rare entity defined as expansion more than 1.5 times of normal coronary artery in diameter. A 77-year-old male was admitted to our hospital with a diagnosis of acute coronary syndrome and a giant right coronary artery aneurysm. Coronary angiography showed occlusion of the right coronary artery and significant stenosis of left anterior descending artery and left circumflex artery. Enhanced computed tomography( CT) showed a right coronary artery aneurysm with a diameter of 38 mm. Urgent coronary artery bypass grafting and coronary artery ligation proximal and distal to the aneurysm were performed. His postoperative course was uneventful.

[Pulmonary Adenocarcinoma in Situ with Positive Findings in Pleural Lavage Cytology].

We report a rare case of positive findings in pleural lavage cytology(PLC) in the patient with pulmonary adenocarcinoma in situ (AIS). A 78-year-old woman was presented with a 30 mm pure groundglass nodule (GGN) in the left upper lobe on chest computed tomography (CT). After 2 years follow- up, thoracoscopic surgery was performed to resect the nodule. PLC was performed before pulmonary resection. Histopathological diagnosis was 25 mm AIS. However, PLC showed positive findings of malignant cells. CT examination at 1 year and 6 months postoperatively showed pleural dissemination findings and the patient died of lung cancer at 3 years and 2 months postoperatively. PLC's contribution to TNM staging has not yet been clarified. The positive findings in PLC and large size of pure GGN were considered likely to be poor prognostic indicators.

DNA methylation analysis of multiple imprinted DMRs in Sotos syndrome reveals IGF2-DMR0 as a DNA methylation-dependent, P0 promoter-specific enhancer.

Haploinsufficiency of NSD1, which dimethylates histone H3 lysine 36 (H3K36), causes Sotos syndrome (SoS), an overgrowth syndrome. DNMT3A and DNMT3B recognizes H3K36 trimethylation (H3K36me3) through PWWP domain to exert de novo DNA methyltransferase activity and establish imprinted differentially methylated regions (DMRs). Since decrease of H3K36me3 and genome-wide DNA hypomethylation in SoS were observed, hypomethylation of imprinted DMRs in SoS was suggested. We explored DNA methylation status of 28 imprinted DMRs in 31 SoS patients with NSD1 defect and found that hypomethylation of IGF2-DMR0 and IG-DMR in a substantial proportion of SoS patients. Luciferase assay revealed that IGF2-DMR0 enhanced transcription from the IGF2 P0 promoter but not the P3 and P4 promoters. Chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) revealed active enhancer histone modifications at IGF2-DMR0, with high enrichment of H3K4me1 and H3 lysine 27 acetylation (H3K27ac). CRISPR-Cas9 epigenome editing revealed that specifically induced hypomethylation at IGF2-DMR0 increased transcription from the P0 promoter but not the P3 and P4 promoters. NSD1 knockdown suggested that NSD1 targeted IGF2-DMR0; however, IGF2-DMR0 DNA methylation and IGF2 expression were unaltered. This study could elucidate the function of IGF2-DMR0 as a DNA methylation dependent, P0 promoter-specific enhancer. NSD1 may play a role in the establishment or maintenance of IGF2-DMR0 methylation during the postimplantation period.

[Coronary Aneurysm with Fistula].

We successfully treated two rare cases of coronary aneurysm with fistula. Case 1;A 65-year-old female referred to our hospital with the coronary aneurysm and fistula. Right coronary aneurysm with fistula leading to coronary sinus was observed. Coronary bypass surgery using a saphenous vein to #4PD was performed, and two right ventricle branches were reconstructed. Coronary aneurysm was resected. Case 2;A 46-year-old male was admitted with chest discomfort. Coronary aneurysm with fistula from the left main trunk to left ventricle was demonstrated. Ligation of the coronary artery aneurysm and suture closure of the entry site to the left ventricle was performed. Both patients had uneventful recovery.

[Third Tricuspid Valve Replacement:Report of a Case].

A 79-year-old man underwent tricuspid valve replacement (TVR) with a bovine pericardial bioprosthesis, Carpentier-Edwards Perimount (CEP), for tricuspid regurgitation 27 years earlier. Twenty-one years after the first operation, he underwent re-TVR with a bovine pericardial bioprosthesis, Magna Mitral Ease, due to prosthetic valve dysfunction. Since the prosthetic valve dysfunction progressed again, the third TVR was performed six years after the second operation. The findings of the resected bioprosthesis showed sclerosis of the septal cusp, and pannus formation between the septal and posterior cusps. For the third TVR, the porcine bioprosthesis Epic was used. Neither transvalvular nor perivalvular leakage was observed postoperatively, and the patient was discharged without any complications.

[Papillary Fibroelastoma Arising from the Left Atrial Wall:Report of a Case].

Papillary fibroelastoma (PFE) is a rare benign cardiac tumor generally arising from the valvular endocardium. We report an extremely rare case of PFE arising from the left atrial wall. A 70-year-old male patient was admitted to our hospital with a diagnosis of left atrial tumor. Echocardiography and enhanced computed tomography showed an approximately 14 mm mass on the left atrial wall. Moreover, the magnetic resonance imaging showed hyperintenseness on a T2-weighted image. We diagnosed the tumor as a myxoma. Intraoperatively, we found a mobile tumor on the left atrial wall. It had a sea anemone-like appearance and was suspected to be PFE. We performed the tumor resection including the left atrial wall. Histological examination confirmed PFE. His postoperative course was uneventful.

PubChemQC PM6: Data Sets of 221 Million Molecules with Optimized Molecular Geometries and Electronic Properties.

We report on optimized molecular geometries and electronic properties calculated by the PM6 method for 94.0% of the 91.6 million molecules cataloged in PubChem Compounds retrieved on August 29, 2016. In addition to neutral states, we also calculated those for cationic, anionic, and spin flipped electronic states of 56.2%, 49.7%, and 41.3% of the molecules, respectively. Thus, the grand total of the PM6 calculations amounted to 221 million. We compared the resulting molecular geometries with B3LYP/6-31G* optimized geometries for 2.6 million molecules. The root-mean-square deviations in bond length and bond angle were approximately 0.016 Å and 1.7°, respectively. Then, using linear regression to examine the HOMO energy levels E(HOMO) in the B3LYP and PM6 calculations, we found that EB3LYP(HOMO) = 0.876EPM6(HOMO) + 1.975 (eV) and calculated the coefficient of determination to be 0.803. Likewise, we examined the LUMO energy levels and found EB3LYP(LUMO) = 1.069EPM6(LUMO) - 0.420 (eV); the coefficient of determination was 0.842. We also generated four subdata sets, each of which was composed of molecules with molecular weights less than 500. Subdata set i contained C, H, O and N, ii contained C, H, N, O, P, and S, iii contained C, H, N, O, P, S, F, and Cl, and iv contained C, H, N, O, P, S, F, Cl, Na, K, Mg, and Ca. The data sets are available at http://pubchemqc.riken.jp/pm6_datasets.html under a Creative Commons Attribution 4.0 International license.

Identification of consensus motifs associated with mitotic recombination and clinical characteristics in patients with paternal uniparental isodisomy of chromosome 11.

Uniparental disomy (UPD) is defined as the inheritance of both homologs of a given genomic region from only one parent. The majority of UPD includes an entire chromosome. However, the extent of UPD is sometimes limited to a subchromosomal region (segmental UPD). Mosaic paternal UPD (pUPD) of chromosome 11 is found in approximately 20% of patients with Beckwith-Wiedemann syndrome (BWS) and almost all pUPDs are segmental isodisomic pUPDs resulting from mitotic recombination at an early embryonic stage. A mechanism initiating a DNA double strand break (DSB) within 11p has been predicted to lead to segmental pUPD. However, no consensus motif has yet been found. Here, we analyzed 32 BWS patients with pUPD by SNP array and searched for consensus motifs. We identified four consensus motifs frequently appearing within breakpoint regions of segmental pUPD. These motifs were found in another nine BWS patients with pUPD. In addition, the seven motifs found in meiotic recombination hot spots could not be found within pUPD breakpoint regions. Histone H3 lysine 4 trimethylation, a marker of DSB initiation, could not be found either. These findings suggest that the mechanism(s) of mitotic recombination leading to segmental pUPD are different from that of meiotic recombination. Furthermore, we found seven patients with paternal uniparental diploidy (PUD) mosaicism. Comparison of clinical features between segmental pUPDs and PUDs showed that developmental disability and cardiac abnormalities were additional characteristic features of PUD mosaicism, along with high risk of tumor development. We also found that macroglossia was characteristic of segmental pUPD mosaicism.

[Pulmonary Non-tuberculous Mycobacteriosis Complicated with Pulmonary Large Cell Neuroendocrine Carcinoma;Report of a Case].

A 64-year-old man with pulmonary non-tuberculous mycobacteriosis(pulmonary NTM) who had been treated by antituberculous chemotherapy, developed a new nodule of 8 mm in size in the segment 3 of the right upper lobe. The cavity of 4.0 cm in size in the segment 1+2 of the left upper lobe due to Mycobacterium avium infection was preexisted. Radiologically, new nodule of the right lung was suspected to be lung cancer. Left upper lobe apical trisegmentectomy was performed at first. Three months later, enlarging of the right lung nodule with increased fluoro-2-deoxy-D-glucose(FDG) activity was noted, and the diagnosis of lung cancer was made by transbronchial lung biopsy(TBLB). Then, right upper lobectomy with systematic nodal dissection were performed.

[A Case of Locally Advanced Sigmoid Colon Cancer Occupying the Pelvis with Successful Total Posterior Pelvic Exenteration after Triplet Chemotherapy].

We report a case of locally advanced colon cancer that directly invaded the rectum wall and uterus resulting in huge mass in the whole pelvis that we could successfully made complete radical resection of the whole tumor without exposing the tumor to the surgical margin after the triplet chemotherapy. The patient was a 57-year-old woman complaining of anus pain, melena, fever, and weight loss. Although swelling of the regional lymph node was observed, no distant metastasis was found resulting in clinical diagnosis of Stage Ⅲb. However, oncologically safe complete resection seemed difficult; thus, chemotherapy( 3 courses of FOLFOX followed by 3 courses of FOLFOXIRI plus bevacizumab)was administered. As a result, significant tumor reduction was observed; therefore, the tumor was completely resected with posterior pelvic exenteration. Final staging was ypT4bypN0M0(ypStage Ⅱ). Eight courses of CapeOX was administered as adjuvant chemotherapy.

[Anterior Mediastinal Myxoid Liposarcoma Detected by Chest Computed Tomogram in the Patient with Pneumothorax].

A 57-year-old woman visited our hospital with left chest pain. Chest computed tomography (CT) scanning showed left pneumothorax with apical bullae and a nodular shadow in the left anterior mediastinum accidentally. However, a week later, we could not detect a mediastinal shadow on chest CT image after healing of left pneumothorax. Video assisted thoracoscopic surgery was scheduled in order to remove bullae and evaluate an anterior mediastinal lesion. The mediastinal lesion was tumorous and resected with around pericardial fat tissue. Pathological diagnosis was a myxoid liposarcoma of 15×10mm in size without infiltration into the surrounding tissue. The postoperative course was uneventful without recurrence 6 months later.

De novo GABRA1 mutations in Ohtahara and West syndromes.

OBJECTIVE: GABRA1 mutations have been identified in patients with familial juvenile myoclonic epilepsy, sporadic childhood absence epilepsy, and idiopathic familial generalized epilepsy. In addition, de novo GABRA1 mutations were recently reported in a patient with infantile spasms and four patients with Dravet syndrome. Those reports suggest that GABRA1 mutations are associated with infantile epilepsy including early onset epileptic encephalopathies. In this study, we searched for GABRA1 mutations in patients with infantile epilepsy to investigate the phenotypic spectrum of GABRA1 mutations. METHODS: In total, 526 and 145 patients with infantile epilepsy were analyzed by whole-exome sequencing and GABRA1-targeted resequencing, respectively. RESULTS: We identified five de novo missense GABRA1 mutations in six unrelated patients. A p.R112Q mutation in the long extracellular N-terminus was identified in a patient with infantile epilepsy; p.P260L, p.M263T, and p.M263I in transmembrane spanning domain 1 (TM1) were identified in three unrelated patients with West syndrome and a patient with Ohtahara syndrome, respectively; and p.V287L in TM2 was identified in a patient with unclassified early onset epileptic encephalopathy. Four of these mutations have not been observed previously. SIGNIFICANCE: Our study suggests that de novo GABRA1 mutations can cause early onset epileptic encephalopathies, including Ohtahara syndrome and West syndrome.

[Papillary Fibroelastoma on the Tricuspid Valve: Report of a Case].

68-year-old male was diagnosed as a cardiac tumor detected by echocardiography, incidentally. He was asymptomatic but the tumor was removed surgically for the diagnostic therapy. It was diagnosed as a papillary fibroelastoma (PFE) clinically and pathologically. He was discharged from the hospital without any complication. PFE on the tricuspid valve is uncommon benign tumor of the heart. Because it is at risk of embolic events, we think it should be operated even if it exists the right side of the heart.

[Stanford Type A Aortic Dissection after Off-pump Coronary Artery Bypass Grafting Using Automated Proximal Anastomotic Device;Report of a Case].

Operative mortality of Stanford type A aortic dissection( AAD) repair with previous cardiac surgery is high, especially with previous coronary artery bypass grafting. We report an extremely rare case of AAD related to the PAS-Port system. A 68-year-old male patient on dialysis was admitted to our hospital with a diagnosis of AAD. Half a year before he had undergone off-pump coronary artery bypass grafting using PAS-Port system for a proximal anastomosis of a vein graft. Urgent graft replacement of the ascending aorta was performed, and the proximal anastomotic site of the vein graft was attached to the prosthetic graft. Intraoperative investigation revealed that an intimal tear was located on the anastomotic site made by the PAS-Port system. The postoperative course was uneventful and he has been visiting our hospital regularly for dialysis.

Risk factors for a persistent type 2 endoleak after endovascular aneurysm repair.

PURPOSE: To investigate the natural course of type 2 endoleaks (T2Es) and to identify the risk factors associated with a persistent T2E after endovascular aneurysm repair (EVAR). METHODS: The medical records of patients who underwent EVAR for the treatment of an atherosclerotic abdominal aortic aneurysm between October 2006 and December 2011 at our institute were reviewed. T2Es were diagnosed by contrast-enhanced computed tomography within 4 weeks of EVAR, and patients were followed up at 6 and 12 months. In cases where a T2E was detected, the blood vessels responsible for the T2E were identified and statistically analyzed for their association with a persistent T2E. RESULTS: We identified T2Es in 111 of 469 patients within 4 weeks of undergoing EVAR. During the follow-up, 41 patients (36.9 %) showed spontaneous resolution of their T2E. The percentage of patients with a T2E was 75.4, 69.2 and 58.0 % at 6, 12 and 24 months, respectively. T2E caused by defects in multiple vessels and T2E associated with the fourth lumbar artery were identified as risk factors associated with a persistent T2E in the univariate analysis. In the multivariate analysis, T2E caused by multiple vessels was identified as the only independent risk factor for a persistent T2E. CONCLUSIONS: We identified T2E caused by multiple vessel failure as an independent risk factor for persistent T2E.

Comprehensive and quantitative multilocus methylation analysis reveals the susceptibility of specific imprinted differentially methylated regions to aberrant methylation in Beckwith-Wiedemann syndrome with epimutations.

PURPOSE: Expression of imprinted genes is regulated by DNA methylation of differentially methylated regions (DMRs). Beckwith-Wiedemann syndrome is an imprinting disorder caused by epimutations of DMRs at 11p15.5. To date, multiple methylation defects have been reported in Beckwith-Wiedemann syndrome patients with epimutations; however, limited numbers of DMRs have been analyzed. The susceptibility of DMRs to aberrant methylation, alteration of gene expression due to aberrant methylation, and causative factors for multiple methylation defects remain undetermined. METHODS: Comprehensive methylation analysis with two quantitative methods, matrix-assisted laser desorption/ionization mass spectrometry and bisulfite pyrosequencing, was conducted across 29 DMRs in 54 Beckwith-Wiedemann syndrome patients with epimutations. Allelic expressions of three genes with aberrant methylation were analyzed. All DMRs with aberrant methylation were sequenced. RESULTS: Thirty-four percent of KvDMR1-loss of methylation patients and 30% of H19DMR-gain of methylation patients showed multiple methylation defects. Maternally methylated DMRs were susceptible to aberrant hypomethylation in KvDMR1-loss of methylation patients. Biallelic expression of the genes was associated with aberrant methylation. Cis-acting pathological variations were not found in any aberrantly methylated DMR. CONCLUSION: Maternally methylated DMRs may be vulnerable to DNA demethylation during the preimplantation stage, when hypomethylation of KvDMR1 occurs, and aberrant methylation of DMRs affects imprinted gene expression. Cis-acting variations of the DMRs are not involved in the multiple methylation defects.