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ABSTRACTCO2-Responsive CCT Protein (CRCT) is a transcription factor regulating the expression of starch synthesis-related genes in the vegetative organs of rice. Among six rice cultivars (Nipponbare, Koshihikari, Habataki, Takanari, Momiroman, Leafstar), the expression of CRCTwas highly correlated with the starch content of leaf sheath in Nipponbare, but not in Koshihikari, Takanari and Momiroman during plant development. Meanwhile, the expressions of starch synthesis-related genes such as ADP-glucose pyrophosphorylase large subunit 1was correlated with CRCTexpression in Nipponbare, Momiroman, and Leafstar, but the correlations were ambiguous in Koshihikari, Habataki, and Takanari. In addition, the slopes of its regression lines in Nipponbare were larger than those cultivars. These results suggest that the influence of CRCTon starch synthesis varies among these cultivars. Additionally, the similar cultivar differences were observed when these gene expressions were analyzed after sucrose treatment in leaf blade. Some mutations in the promoter regions of the starch synthesis-related genes were identified among these cultivars. There were cultivars in which the presence of mutations correlated with decreased expression of starch synthesis genes in response to CRCT. Thus, these mutations can partially explain the responsiveness of the starch synthesis-related genes to CRCT, and difference in starch content of vegetative organs among rice cultivars.

ABSTRACTCO2-Responsive CCT Protein (CRCT) is suggested to be a positive regulator of starch synthesis-related genes in the vegetative organs of rice. To investigate the physiological function of CRCT, we produced CRCT knockout lines by CRISPR/Cas9 system. CRCT knockout tended to slightly reduce photosynthetic performance and had some adverse effects on growth parameters such as tiller number and straw dry weight. The starch content in the leaf sheath of CRCT knockout lines was significantly lower than that of wild-type. qRT-PCR and RNA-seq analysis showed that sucrose treatment induced the expression of CRCT, which, in turn, induced many starch synthesis-related genes in wild-type, whereas this induction did not occur in CRCT knockout lines. In conclusion, our results suggest that sugar induction of starch synthesis-related genes is almost fully dependent on CRCT, making it a necessary factor for normal photosynthesis and growth of rice.

Pain is perceived not only by personal experience but also vicariously. Pain empathy is the ability to share and understand other’s intentions and emotions in their painful conditions, which can be divided into cognitive and emotional empathy. It remains unclear how centrally acting analgesics would modulate brain activity related to pain empathy and which component of pain empathy would be altered by analgesics. In this study, we examined the effects of the analgesic tramadol on the brain activity for pain empathy in healthy adults. We used 2 tasks to assess brain activity for pain empathy. In experiment 1, we used a well-established picture-based pain empathy task involving passive observation of other’s pain. In experiment 2, we developed a novel pain empathy task to assess brain activity during cognitive and emotional empathy for pain separately in a single task. We conducted a double-blind, placebo-controlled within-subject crossover study with functional magnetic resonance imaging for 33 participants in experiment 1 and 31 participants in experiment 2, respectively. In experiment 1, we found that tramadol decreased activation in the supramarginal gyrus during observation of other's pain compared with placebo. Supramarginal gyrus activation correlated negatively with the thermal pain threshold. In experiment 2, we found that tramadol decreased activation in angular gyrus in cognitive empathy for pain compared with placebo but did not change brain activity in emotional empathy for pain.

ABSTRACTPartial submergence of Oryza sativadeepwater rice elicits enhancement of internodal elongation, referred to as deepwater response, conferred by three types of genes, SNORKEL1/2(SK1/2), SEMIDWARF1(SD1), and ACCELERATOR OF INTERNODE ELONGATION 1(ACE1). We investigated the presence and expression of these genes in the African cultivated rice Oryza glaberrimaand the relationship between these genes and the deepwater response of O. glaberrima. In 49 of the 50 accessions tested, one or two SKgenes were identified, which could be divided into three types of SK1and four types of SK2. The accessions with the SK2type whose expression was induced by submergence demonstrated rapid internodal elongation under submergence. In most of these accessions, submergence also increased the expression of SD1and ACE1genes. However, the accessions did not possess the haplotype of SD1that is associated with high deepwater response in O. sativa. In contrast, they possessed the type of ACE1gene similar to that in O. sativadeepwater rice. These results indicate that the molecular mechanisms underlying induction of deepwater response in O. glaberrimaare similar to that found in deepwater rice of O. sativaand suggest that most O. glaberrimacultivars, including upland cultivars, can exhibit rapid internodal elongation under submergence.

ABSTRACTIn the present study, 15 accessions of wild rice species belonging to genome groups different from that of Oryza sativawere tested for their adaptive response to flooding at the seedling and mature stages. Under complete submergence at the seedling stage, reduced underwater shoot elongation and high survival rate after the recovery period were observed in the accessions of O. minutaas well as tetraploid O. punctata, O. eichingeri, O. officinalis, O. alta, O. grandiglumis, O. latifolia, and O. australiensis. This suggests that these species exhibit submergence tolerance at the seedling stage. During gradual submergence at mature stage, promoted internodal elongation was observed in the accessions of diploid and tetraploid O. punctata, O. alta, O. grandiglumis, O. latifolia, and O. brachyantha. This suggests that these species possess floating ability. Remarkably, two CCDD genome species, namely O. grandiglumisand O. latifolia, strongly displayed both submergence tolerance at the seedling stage and floating ability at mature stage, suggesting that these species are valuable genetic resources for improving rice adaptability to flooding. SUB1Aand SNORKELgenes are known to confer submergence tolerance and floating ability to O. sativa, respectively. However, SUB1A, SNORKEL1, and SNORKEL2genes were not detected in any of the wild rice accessions investigated in the present study. Our results suggest that adaptive responses to flooding in the investigated wild rice species can be achieved independently of the presence or absence of SUB1Aand SNORKELgenes.

This paper introduces a new framework for singer diarization, which is a technique to reveal who sings when in songs with multiple singers. Although various techniques have been developed to analyze and extract features of singing voices in musical audio signals, most of them assume that a song is sung by a single singer, and singer diarization for multiple singers has not been well studied in the field of singing information processing. To deal with multiple speakers in speech analysis, speaker diarization has been explored to handle overlapped speech voices, but cannot handle singing voices well because of acoustic differences between singing and speech voices. This paper therefore proposes a new diarization framework specialized in singing voices. To achieve high accuracy in overlap detection, this paper proposes a novel acoustic feature named Cosacorr score, which is helpful in estimating whether a song is sung by more than one singer. After extracting singing voices from polyphonic music by using a singing voice separation technique, the framework adopts an existing ArcFace technique to extract discriminative singer representations from short segments of the separated singing voices. The framework is evaluated by using a new private dataset of unison singing voices, which is constructed using commercially available compact discs (CDs). The experimental results show that the proposed framework outperformed the baseline method for speaker diarization in terms of diarization error rate (DER).

ABSTRACTWe screened 80 Asian rice (Oryza sativaL.) cultivars for the presence of the submergence-tolerance gene SUB1A-1and the floating genes SNORKEL1(SK1) and SNORKEL2(SK2), and found that the deepwater rice cultivar Baisbish (BSB) and the submergence-tolerant cultivar Flood Resistant 13A (FR13A) both possess the SUB1A-1and the SK1/2. When BSB and FR13A seedlings were completely submerged, spindly growth of shoots was induced in BSB but not in FR13A. Submergence significantly increased the SUB1A-1transcript abundance in BSB and FR13A shoots, but the expression level in BSB was much lower than that of FR13A. Submergence also induced the expression of both ERF66and ERF67, the transcriptional targets of SUB1A-1, in FR13A shoots, whereas it upregulated the expression of ERF67but not that of ERF66in BSB shoots. These results suggest that BSB could not display submergence tolerance due to the low expression of SUB1A-1and/or ERF66under submergence.

Background. Some cutaneous T-cell lymphoproliferative diseases (LPD), such as lymphomatoid papulosis and pityriasis lichenoides et varioliformis acuta, are characterized by an indolent or waning and waxing clinical course. However, such T-cell LPD are rarely documented in other organs. Methods. A patient with T-cell LPD of the digestive tract characterized by repetitive episodes of self-healing ulcers in the oral and intestinal mucosa over the course of 17 years is reported. Biopsy specimens from oral and intestinal mucosa were studied by conventional pathology, immunocytochemistry, and Southern blot analysis of T-cell receptor (TCR)beta and -gamma gene rearrangement. Results. Immunocytochemically, the infiltrating lymphocytes were lamina propria T cells with a dominant phenotype CD3+, CD4+/-, CD8-, and HML-1-. DNA study revealed the same rearranged configuration of TCR-beta and -gamma genes in specimens from both oral and colonic lesions. Conclusions. The present case may represent a novel T-cell lymphoproliferative disease (i.e., a digestive-tract mucosal counterpart of cutaneous dysplastic LPD). Cancer 1995;75:1728-33.

Background. Mutations in the K-ras oncogene at codon 12 are detected at a remarkably high frequency in pancreatic carcinomas and are believed to be a critical event in oncogenesis. The authors attempted to detect Kras mutations in DNA obtained from pure pancreatic juice collected endoscopically, as a novel diagnostic approach to pancreatic carcinoma. Methods. K-ras mutations were examined using the two-step polymerase chain reaction (PCR) combined with restriction enzyme digestion, followed by nonradioisotopic single-strand conformation polymorphism (SSCP) analysis. Results. Specific mutations of the K-rus gene at codon 12 were found in six of nine (67%) duct cell carcinomas, all of which were negative by cytodiagnosis of the same pure pancreatic juice. K-ras mutations were not detected in the pancreatic juice from 14 healthy control subjects, 10 patients with chronic pancreatitis, or 3 patients with islet cell tumors. Conclusions. Detection of K-ras mutation at codon 12 in pancreatic juice is highly specific for diagnosing pancreatic duct cell carcinoma and may be a valuable diagnostic modality for pancreatic carcinoma and for differentiating chronic pancreatitis from carcinoma. Cancer 1994; 73:1589-94. Key words: K-ras oncogene, pancreatic carcinoma, endoscopic retrograde pancreatography, pancreatic juice, single-strand conformation polymorphism, polymerase chain reaction.

Background. The molecular genetic changes associated with colorectal carcinoma are among the best understood of any common human cancer. The genetic changes during the late stages of colorectal carcinomas may be useful in clinical management for determining the metastatic potential of the carcinoma. Methods. Tumor tissues were evaluated by restriction fragment length polymorphism (RFLP) analysis of chromosomes 5q, 17p, 18q, and 22q (n = 98), by reverse transcription-polymerase chain reaction (RT-PCR) analysis of messenger RNA expression of the DCC gene (deleted in colorectal carcinoma) (n = 27) and by immunohistochemical analysis of p53 protein expression (n = 44). Results. Loss of heterozygosity (LOH) on chromosomes 17p, 18q, and 22q, but not on 5q, was much more frequently detected in advanced carcinomas than in intramucosal carcinomas (P < 0.01). 17p LOH was significantly correlated with vascular invasion (P < 0.001), whereas 18q LOH was correlated with lymphatic invasion and hepatic metastasis (P < 0.01), and 22q LOH was correlated with lymph node metastasis (P < 0.05). LOH on 5q did not show a significant correlation with any factors of tumor invasion or metastasis. DCC expression was not observed in any of five hepatic metastasis or in five of seven advanced carcinomas that were accompanied by hepatic metastasis (10 of 12). However, a similar lack of expression was observed in only 5 of 15 carcinomas without hepatic metastasis (P < 0.05). p53 Expression was found to vary in both primary and metastatic carcinomas by immunohistochemistry. Conclusions. The clinical application of molecular genetics (i.e., RFLP analysis of chromosome 17p, 18q, and 22q and RT-PCR analysis of DCC expression into messenger RNA) can be used to determine the metastatic potential of colorectal carcinomas. Cancer 1994; 73:1324-31.

Background. Pyothorax-associated pleural lymphomas (PAPL) occur in patients with long-standing pyothorax. Epstein-Barr virus (EBV) plays an important role in its development. Methods. An atypical case of PAPL is reported and its clinical features are compared with those of five cases of ordinary PAPL. Results. The histology of the lymphoma changed from prominent T-cell infiltration to an overt B-cell lymphoma of diffuse, large cell-type. DNA studies of the lymphoma at the first presentation demonstrated dual rearrangement in the T-cell receptor [beta] (TCR,[beta]) and immunoglobulin heavy chain J region (IgJH) genes. EBV genome analysis demonstrated monoclonal expansion of EBV-infected cells. EBV gene products were present onlv in large lymphocytes with B-cell phenotype. The same rearranged band of the IgJH gene, but not clonal rearrangement of the TCR[beta] gene, was observed in the overt B-cell lymphoma at the relapse. The same EBV-infected clone was observed throughout the course of the disease. The tumor was localized at the pyothorax wall, and cell-mediated immunity, reflected by a positive tuberculin reaction, was preserved only in the current case. Conclusions. T-cell-rich lymphoid infiltration in the current case is a variant of EBV-associated PAPL, which is infiltrated by reactive T-cells with clonal accentuation. This is the first documented case of Ebv-associated PAPL evolving from T-cell-rich lymphoid infiltration.

ABSTRACTRubisco is the key enzyme responsible for photosynthetic CO2fixation. Previously, we succeeded to increase the catalytic rate (kcat) of Rubisco by the overexpression of sorghum C4-type Rubisco small subunit (RbcS) in rice. These Rubisco are present as a chimera of sorghum RbcS and rice RbcS. In this study, we introduced an RNAi construct for rice RbcSinto a sorghum RbcS overexpression line (SS10). Obtained double-transgenic lines iSS4 and iSS5 dominantly expressed sorghum RbcS and accumulated 35% and 20% of Rubisco content of non-transgenic rice (WT), respectively. Rubisco in these double-transgenic lines showed higher kcatand Kmfor CO2than WT and SS10. Moreover, iSS4 showed higher photosynthetic nitrogen use efficiency under high CO2conditions. These results demonstrate the usefulness of C4-type RbcS to improve photosynthetic performance under elevated CO2conditions.

Epstein–Barr virus (EBV) is associated with several human malignancies including 8–10% of gastric cancers (GCs). Genome-wide analysis of 3D chromatin topologies across GC lines, primary tissue and normal gastric samples revealed chromatin domains specific to EBV-positive GC, exhibiting heterochromatin-to-euchromatin transitions and long-range human–viral interactions with non-integrated EBV episomes. EBV infection in vitro suffices to remodel chromatin topology and function at EBV-interacting host genomic loci, converting H3K9me3+heterochromatin to H3K4me1+/H3K27ac+bivalency and unleashing latent enhancers to engage and activate nearby GC-related genes (for example TGFBR2and MZT1). Higher-order epigenotypes of EBV-positive GC thus signify a novel oncogenic paradigm whereby non-integrative viral genomes can directly alter host epigenetic landscapes (‘enhancer infestation’), facilitating proto-oncogene activation and tumorigenesis.

We report an extremely rare case of lymphoepithelioma-like carcinoma of the ureter. A 73-year-old man presented with asymptomatic gross hematuria. Based on the clinical diagnosis of a left ureteral tumor, the patient underwent left nephroureterectomy. Histological examination showed lymphoepithelioma-like carcinoma of the ureter with a unique growth pattern of submucosal spread, in which cytokeratin immunohistochemistry clearly demonstrated the cluster of neoplastic cells within the dense lymphocytic infiltrates. The tumor cells showed negative results for the presence of Epstein-Barr virus by Epstein-Barr virus-encoded ribonucleic acid in situ hybridization. The patient was free of tumor recurrence and metastasis 15 months after the operation. To avoid misdiagnosis as chronic inflammation, recognition of this type of tumor is important. Keywords: lymphoepithelioma-like carcinoma | ureter | immunohistochemistry | Epstein-Barr virus | in situ hybridization, Introduction Lymphoepithelioma is an undifferentiated carcinoma with intense lymphoid infiltrates arising in the nasopharynx. (1) Carcinomas that histologically resemble lymphoepithelioma of the nasopharynx are called lymphoepithelioma-like carcinoma (LELC). Only five [...]

Gastric neoplasms exhibiting oxyntic gland differentiation typically are composed of cells with mild cytonuclear atypia differentiating to chief cells and to a lesser extent, parietal cells. Such tumors with atypical features have been reported also and terminology for this entity remains a matter of considerable debate. We analyzed and classified 26 tumors as oxyntic gland neoplasms within mucosa (group A, eight tumors) and with submucosal invasion. The latter was divided further into those with typical histologic features (group B, 14 tumors) and atypical features, including high-grade nuclear or architectural abnormality and presence of atypical cellular differentiation (group C, four tumors). Groups A and B tumors shared similar histologic features displaying either a chief cell predominant pattern characterized by monotonous chief cell proliferation, or a well-differentiated mixed cell pattern showing admixture of chief and parietal cells resembling fundic gland. In addition, group C tumors displayed atypical cellular differentiation, including mucous neck cell and foveolar epithelium. Moderate or even marked cytological atypia was noted in group C, whereas it was usually mild in the other groups except for three group B tumors with focal moderate atypia. More than 1000 μm submucosal invasion and lymphovascular invasions were recognized only in group C. Mutation analyses identified KRASmutation in one group C tumor as well as GNASmutation in in one group A and group B tumors. Intramucosal tumors appear to behave biologically benign and should be classified as “oxyntic gland adenoma”. Those with submucosal invasion also have low malignant potential; however, a subset will have atypical features associated with aggressive histologic features and should be designated as “adenocarcinoma of fundic gland type”. Especially, we suggest “adenocarcinoma of fundic gland mucosa type” for tumors with submucosal invasion exhibiting atypical cellular differentiation, because the feature is likely to be a sign of aggressive phenotype.

Malignancies occur in approximately 1:1000 pregnancies; the most common being breast (46%) and hematological (18%) malignancies. Oral cancers account for only 2% of all cancers in pregnant women, and there are no standard guidelines for the treatment of oral cancer during pregnancy. Between 2007 and 2014, our department managed 1109 patients with oral cancers; four (0.4%) had tongue carcinomas during pregnancy. These cases were retrospectively reviewed. The four women were aged 29–39 (median 32.5) years. Two underwent partial glossectomy at 39 and 40 weeks’ gestation, respectively, one received radiotherapy at 17 weeks’ gestation, and one underwent supraomohyoid neck dissection and hemi-glossectomy with a forearm flap reconstruction. In addition to tumor factors, the wishes of the patient and her family, gestational age, and fetal and maternal conditions are important factors in deciding on a treatment protocol. Moreover, treatment decisions require multidisciplinary approach.

Supplemental Digital Content is available in the text.Intrahepatic cholangiocarcinoma (ICC) is an extremely aggressive carcinoma. Useful predictors for the patients’ prognosis after surgery have not been fully established. From the University of Tokyo Hospital pathology archives, we reviewed 107 cases of ICC, 54 cases of perihilar cholangiocarcinoma, and 40 cases of extrahepatic cholangiocarcinoma (ECC); we also investigated the significance of tumor budding in ICC, in comparison with perihilar cholangiocarcinoma and ECC. The tumor-budding frequencies were different by tumor location: 40.2% (43/107) in ICC, 70.4% (38/54) in perihilar cholangiocarcinoma, and 60.0% (24/40) in ECC. Tumor budding in ICC was associated with many pathologic indicators associated with invasion, such as major vascular invasion (P=0.012) and Union for International Cancer Control stage (P=0.007). Univariate and multivariate Cox regression analyses revealed tumor budding as a powerful prognostic factor for both recurrence-free survival (RFS) and overall survival (OS) in ICC by univariate (RFS: hazard ratio [HR]: 2.666; 95% confidence interval [CI]: 1.517-4.683, OS: HR: 4.206; 95% CI: 2.447-7.230) and by multivariate analyses (RFS: HR: 3.038; 95% CI: 1.591-5.973, OS: HR: 4.547, 95% CI: 2.348-8.805). Tumor budding was also a significant prognostic factor of perihilar cholangiocarcinoma, but not of ECC. When ICC was divided into 2 subtypes, type 1 (hilar) and type 2 (peripheral), tumor budding was the strong prognostic factor in type 2 ICC, but not in type 1 ICC, suggesting that some differences in biological behavior exist between type 1 ICC and perihilar cholangiocarcinoma. Tumor budding is prognostically important in ICC, and its pathogenetic role in biliary tract carcinomas might be different by anatomic location.

ABSTRACTOverexpression of Rubisco small subunit (RbcS) of C4plant, sorghum (sorghum bicolor) was shown to enhance the catalytic turnover rate (kcat) of Rubisco in rice (Oryza sativa). In this study, the effects of other Rubisco small subunits of C4plants, Napier grass (Pennisetum purpureum) and guinea grass (Megathyrsus maximus) on kinetic properties of Rubisco in rice were studied. The expression levels of Napier grass RbcS(NgRbcS) and guinea grass RbcS(GgRbcS) proteins accounted for 41% and 45% of total RbcS, respectively in homozygous overexpression lines. The kcatand Kmfor CO2(Kc) of Rubisco were increased in all transgenic lines. Interestingly, the kcatwas markedly higher in NgRbcS homozygous line, whereas Kcwas notably higher in GgRbcS homozygous line. Although its effects depend on species, these results suggest that the introduction of C4RbcS are effective approaches to alter the catalytic properties of Rubisco in rice.AbbreviationGgRbcS: guinea grass RbcS; Kc: Kmfor CO2; kcat: catalytic turnover rate; NpRbcS: Napier grass RbcS; RbcL: Rubisco large subunit; RbcS: Rubisco small subunit; Sc/o: CO2/O2specificity.

Dedifferentiated chondrosarcoma is a rare bone sarcoma, whose genetic background remains incompletely understood. Mutations in SUZ12or EED, which encode polycomb repressive complex 2 (PRC2) components, and resulting deficiency in H3K27me3 are characteristic features of the majority of malignant peripheral nerve sheath tumors. Here, we investigated H3K27me3 and PRC2 status in dedifferentiated chondrosarcoma. Among 19 evaluable dedifferentiated chondrosarcoma cases, six (32%) showed immunohistochemical loss of H3K27me3 only in the dedifferentiated component, whereas the well-differentiated component retained H3K27me3. H3K27me3-deficient dedifferentiated chondrosarcoma occurred in two men and four women with a median age of 66. All of these tumors affected bones of the upper half of the body, with the ribs being preferentially involved, which represented a significantly different distribution compared to that in the 13 H3K27me3-intact dedifferentiated chondrosarcomas. H3K27me3-deficient dedifferentiated chondrosarcomas were histologically different from H3K27me3-intact dedifferentiated chondrosarcomas, as the former invariably demonstrated dedifferentiated histology with a striking similarity to classic malignant peripheral nerve sheath tumor, comprising sweeping to swirling fascicles of relatively uniform spindle cells. Heterologous rhabdomyoblastic differentiation, the focal presence of grade 3 chondrosarcoma histology, and a cartilaginous component in the metastatic sites were exclusively seen in some cases of H3K27me3-deficient dedifferentiated chondrosarcoma. In all three H3K27me3-deficient dedifferentiated chondrosarcomas that contained focal grade 3 histology, dedifferentiated components did not juxtapose to the grade 3 areas but transitioned abruptly from the grade 1–2 components. Targeted next generation sequencing, which was successfully performed on four H3K27me3-deficient dedifferentiated chondrosarcomas, identified an IDH2mutation in one case and COL2A1truncations in three cases. The dedifferentiated areas of three cases harbored SUZ12or EEDalterations, which were absent in the well-differentiated component, suggesting a role for PRC2 aberrations in dedifferentiation. H3K27me3 deficiency defines a novel subset of dedifferentiated chondrosarcoma that requires recognition because of its diagnostic and potential clinical implications.

•Liver attenuation on PMCT was compared with clinical, AMCT and autopsy findings.•PMCT may reflect liver pathological change around death.•Herein, assessment of decreased liver attenuation on PMCT needed autopsy findings.

Metastases to the colon are rare and a high-frequency primary region is the stomach. In cases of metastases to the colon, the morphological type of the metastatic region is mostly the infiltrating type of poorly differentiated or undifferentiated adenocarcinoma with lymph and blood vessel invasion. A case of cancer metastasis to the transverse colon that originated from advanced gastric cancer, which shows the difficulties in the precise diagnosis of metastases to the colon, is presented. In the present case, the gastric carcinoma was determined to be an advanced infiltrative ulcerative adenocarcinoma and the colon carcinoma was determined to be a superficial depressed adenocarcinoma. After surgery, the colon carcinoma was diagnosed as a metastatic adenocarcinoma from gastric adenocarcinoma with high invasion of vessels, by immunohistopathological analysis of CK7, CK20, p53 and HER-2. In this report, previously reported cases of metastases to the colon from gastric cancer were reviewed and their morphological characteristics were analyzed.

We have developed an activatable red fluorescence probe for dipeptidylpeptidase-IV (DPP-IV) by precisely controlling the photoinduced electron transfer (PeT) process of a red fluorescent scaffold, SiR600. The developed probe exhibited an extremely low background signal and showed significant fluorescence activation upon reaction with DPP-IV, enabling sensitive detection of esophageal cancer in clinical specimens from cancer patients.

ABSTRACTSugar metabolism pathways such as photosynthesis produce dicarbonyls, e.g. methylglyoxal (MG), which can cause cellular damage. The glyoxalase (GLX) system comprises two enzymes GLX1 and GLX2, and detoxifies MG; however, this system is poorly understood in the chloroplast, compared with the cytosol. In the present study, we determined GLX1 and GLX2 activities in spinach chloroplasts, which constituted 40% and 10%, respectively, of the total leaf glyoxalase activity. In Arabidopsis thaliana, five GFP-fusion GLXs were present in the chloroplasts. Under high CO2concentrations, where increased photosynthesis promotes the MG production, GLX1 and GLX2 activities in A. thalianaincreased and the expression of AtGLX1-2and AtGLX2-5was enhanced. On the basis of these findings and the phylogeny of GLX in oxygenic phototrophs, we propose that the GLX system scavenges MG produced in chloroplasts during photosynthesis.

We report a case of difficult nasal intubation utilizing a Pentax-Airway scope®AWS-100. A 4-month-old female with a rapidly growing melanotic neuroectodermal tumor was scheduled for resection under general anesthesia. The tumor was a large rubbery mass located in the middle of the mandible. For nasal intubation using the AWS, guidance of the tube toward the glottis was attempted using pediatric Magill forceps. Although we could hold the tube with the Magill forceps, it was difficult to insert the tube into the trachea due to the limited space in her hypopharynx. We then used a standard laryngoscope with a Miller straight blade for direct visual laryngoscopy and successfully intubated the patient with the aid of the pediatric Magill forceps. We often experience difficulty navigating a nasal endotracheal tube toward the glottis even when a clear glottic view is obtained with video laryngoscopes, especially in children with a small oropharyngeal space. However, some reports have been shown that video laryngoscopes are useful for intubation of the difficult airway and causes less stress to the upper airway than direct visual laryngoscopy. Video laryngoscopy can be an excellent way to provide nasal intubation in some but not all children.

Pancreatic masses consisting of lipomatous components clinically include lipoma, liposarcoma, lipomatous pseudohypertrophy of the pancreas, fat-containing neoplasms such as perivascular epithelioid cell tumor, and malignant neoplasm with lipoid degeneration. We present pancreatic lipomatous hamartoma, which has not been reported hitherto. A solid pancreatic mass was detected from a computed tomographic scan check-up in each of 3 cases of Japanese men. Macroscopically, well-demarcated solid lipomatous masses were detected at the uncus, body, and tail of the pancreas, respectively. Microscopically, the masses predominantly consisted of mature adipocytes with no atypia, but contained characteristics components of pancreatic hamartoma, such as small ducts, a well-preserved acinar structure, and/or fibrous stroma. On the basis of the unique features, lack of islets and absence of periductal elastic fibers, these tumors are a distinct variant of pancreatic hamartoma. Furthermore, high-mobility group AT-hook 2 expression in the fibro-adipocytes of this tumor indicated that these cells are an integral component of the pancreatic lipomatous hamartoma. Consequently, the unique tumors described herein are pancreatic lipomatous hamartoma, which must be discriminated from other lipomatous lesions of the pancreas.

Tumor invasion is a critical first step in the organismic dissemination of cancer cells and the formation of metastasis in distant organs, the most important prognostic factor and the actual cause of death in most of the cancer patients. We report herein that the cell surface protein podoplanin (PDPN), a potent inducer of cancer cell invasion, is conspicuously expressed by the invasive front of squamous cell carcinomas (SCCs) of the cervix in patients and in the transgenic human papillomavirus/estrogen mouse model of cervical cancer. Laser capture microscopy combined with gene expression profiling reveals that the expression of interferon-responsive genes is up-regulated in PDPN-expressing cells at the tumor invasive front, which are exposed to CD45-positive inflammatory cells. Indeed, PDPN expression can be induced in cultured SCC cell lines by single or combined treatments with interferon-γ, transforming growth factor-β, and/or tumor necrosis factor-α. Notably, shRNA-mediated ablation of either PDPN or STAT1 in A431 SCC cells repressed cancer cell invasion on s.c. transplantation into immunodeficient mice. The results highlight the induction of tumor cell invasion by the inflammatory cytokine–stimulated expression of PDPN in the outermost cell layers of cervical SCC.

Supplemental Digital Content is available in the text.Malignant peripheral nerve sheath tumor (MPNST) and dedifferentiated liposarcoma (DDLPS) are 2 major types of pleomorphic spindle cell sarcoma. The differentiation of MPNST and DDLPS by histomorphology alone can be problematic. Although MDM2amplification and PRC2 alteration leading to H3K27me3 deficiency are genetic hallmarks of DDLPS and MPNST, respectively, a small number of MDM2-amplified MPNSTs and H3K27me3-deficient DDLPSs have been reported in the literature. We systematically compared MDM2 and H3K27me3 status in 68 MPNSTs and 47 DDLPSs. Of the 62 MPNSTs, 22 were immunopositive for MDM2, mostly in a weak and/or focal manner. Of the 21 MDM2-positive MPNSTs successfully tested by fluorescence in situ hybridization, high-level MDM2amplification was observed in 1 case. In contrast, MDM2 staining and high-level MDM2amplification were positive in all the DDLPS tested (28/28 and 20/20). Of the 68 MPNSTs, 42 cases (62%) exhibited complete loss of H3K27me3. All the 13 MPNSTs that showed heterologous differentiation were deficient in H3K27me3. Of the 47 DDLPSs, 3 cases (6%) had complete loss of H3K27me3, all of which exhibited heterologous differentiation. One case of H3K27me3-deficient DDLPS exhibited homozygous loss of EEDaccording to targeted next-generation sequencing, whereas there were no alterations in NF1and CDKN2A. In conclusion, high-level MDM2amplification strongly suggests DDLPS over MPNST. Although a good marker for MPNST, H3K27me3 deficiency also uncommonly occurs in DDLPS in association with PRC2 mutational inactivation. Because both markers are imperfectly specific, rare sarcomas with dual features could be encountered, and their classification should integrate other parameters.

To assess lung respiratory movement (“lung sliding”) in dogs using B‐mode ultrasonography (US) and to develop a method that assesses adhesions between the parietal pleura and the lung. Seventeen male beagles were anesthetized, and respiratory management was performed with intermittent positive pressure ventilation. Lung‐sliding assessments and adhesion examinations were performed with lung US under general anesthesia before and 2 weeks after thoracotomy. Lung sliding was scored on a 4‐level scale based on the percentage of the area that showed lung sliding (3, an area of roughly ≥80% of the intercostal space; 2, about 50% of the area of the intercostal space; 1, a small area of the intercostal space; or 0, movement absent); scores of 0, 1, and 2 indicated adhesions, whereas a score of 3 indicated no adhesions. The animals were then euthanized, and necropsy was performed to examine pleural adhesions. Lung US and necropsy findings were compared. The median lung‐sliding score for the 12 sites with pleural adhesions on necropsy was 1.5, whereas it was 3.0 for the 532 sites without pleural adhesions. The lung‐sliding score was significantly lower in the group with adhesions (P< .0001). Adhesion sites detected on necropsy were in accordance with the sites that had decreased lung‐sliding scores. Lung US could detect pleural adhesions with sensitivity of 100.0% and specificity of 87.8%. Examination of lung sliding by thoracic US has high diagnostic value for detecting canine pleural adhesions and is useful in predicting adhesion sites before thoracic surgery in healthy dogs.

Supplemental Digital Content is available in the text.

ObjectiveMetabolic reprogramming of tumour cells that allows for adaptation to their local environment is a hallmark of cancer. Interestingly, obesity-driven and non-alcoholic steatohepatitis (NASH)-driven hepatocellular carcinoma (HCC) mouse models commonly exhibit strong steatosis in tumour cells as seen in human steatohepatitic HCC (SH-HCC), which may reflect a characteristic metabolic alteration.DesignNon-tumour and HCC tissues obtained from diethylnitrosamine-injected mice fed either a normal or a high-fat diet (HFD) were subjected to comprehensive metabolome analysis, and the significance of obesity-mediated metabolic alteration in hepatocarcinogenesis was evaluated.ResultsThe extensive accumulation of acylcarnitine species was seen in HCC tissues and in the serum of HFD-fed mice. A similar increase was found in the serum of patients with NASH-HCC. The accumulation of acylcarnitine could be attributed to the downregulation of carnitine palmitoyltransferase 2 (CPT2), which was also seen in human SH-HCC. CPT2 downregulation induced the suppression of fatty acid β-oxidation, which would account for the steatotic changes in HCC. CPT2 knockdown in HCC cells resulted in their resistance to lipotoxicity by inhibiting the Src-mediated JNK activation. Additionally, oleoylcarnitine enhanced sphere formation by HCC cells via STAT3 activation, suggesting that acylcarnitine accumulation was a surrogate marker of CPT2 downregulation and directly contributed to hepatocarcinogenesis. HFD feeding and carnitine supplementation synergistically enhanced HCC development accompanied by acylcarnitine accumulation in vivo.ConclusionIn obesity-driven and NASH-driven HCC, metabolic reprogramming mediated by the downregulation of CPT2 enables HCC cells to escape lipotoxicity and promotes hepatocarcinogenesis.

AbstractProduction of bioethanol from rice straw has attracted attention from the point of effective use of agricultural residue. Starch content is an important determinant for bioethanol production from rice straw. The overexpression of CO2-responsive CCT protein(CRCT), which is the positive regulator of starch synthesis in vegetative organs, notably increased the starch content in rice straw. To produce the bioethanol from rice straw, the dilute acid pretreatment is a general pretreatment method. Importantly, the glucose yields in liquid hydrolyzate after dilute acid pretreatment was markedly increased in the CRCToverexpression lines compared with non-transgenic rice. In addition, the overexpression of CRCTenhanced the biomass production. In contrast, CRCTdid not affect on the glucose yields from cellulose in acid-insoluble residue obtained after dilute acid pretreatment. With respect to byproduct in liquid hydrolyzate which inhibits the fermentation, the formic acid content was increased, whereas the furfural, 5-hydroxymethylfurfural and acetic acid contents were unchanged by the overexpression of CRCT. These results demonstrate that genetic engineering of CRCTis an effective method to increase the bioethanol production from rice straw.

Recent successes in tumor immunotherapies have highlighted the importance of tumor immunity. However, most of the work conducted to date has been on T cell immunity, while the role of B cell immunity in cancer remains more elusive. In this study, immunogenetic repertoire profiling for tumor-infiltrating B and T cells in gastric cancers was carried out to help reveal the architecture of B cell immunity in cancer. Humoral immunity in cancer was shown to involve oligoclonal expansions of tumor-specific and private B cell repertoires. We find that B cell repertoires in cancer are shaped by somatic hypermutation (SHM) either with or without positive selection biases, the latter of which tended to be auto-reactive. Importantly, we identified sulfated glycosaminoglycans (GAGs) as major functional B cell antigens among gastric tumors. Furthermore, natural anti-sulfated GAG antibodies discovered in gastric cancer tissues showed robust growth-suppressive functions against a wide variety of human malignancies of various organs.

A primitive cell-like gene expression signature is associated with aggressive phenotypes of various cancers. We assessed the expression of phenotypic markers characterizing primitive cells and its correlation with clinicopathologic and molecular characteristics in gastric cancer. Immunohistochemical analysis of a panel of primitive phenotypic markers, including embryonic stem cell markers (OCT4, NANOG, SALL4, CLDN6, and LIN28) and known oncofetal proteins (AFP and GPC3), was performed using tissue microarray on 386 gastric cancers. On the basis of the expression profiles, the 386 tumors were clustered into 3 groups: group 1 (primitive phenotype, n=93): AFP, CLDN6, GPC3, or diffuse SALL4 positive; group 2 (SALL4-focal, n=56): only focal SALL4 positive; and group 3 (negative, n=237): all markers negative. Groups 1 and 2 predominantly consisted of intestinal-type adenocarcinoma, including 13 fetal gut-like adenocarcinomas exclusively in group 1. Group 1 was significantly associated with higher T-stage, presence of vascular invasion and nodal metastasis when compared with groups 2 and 3. Group 1 was associated with patients’ poor prognosis and was an independent risk factor for disease-free survival. Group 1 showed frequent TP53 overexpression and little association with Epstein-Barr virus or mismatch repair deficiency. Further analysis of the Cancer Genome Atlas data set validated our observations and revealed that tumors with primitive phenotypes were mostly classified as “chromosomal instability” in the Cancer Genome Atlas’ molecular classification. We identified gastric cancer with primitive enterocyte phenotypes as an aggressive subgroup of intestinal-type/chromosomal instability gastric cancer. Therapeutic strategies targeting primitive markers, such as GPC3, CLDN6, and SALL4, are highly promising.

Supplemental Digital Content is available in the text.

Cancer cells use PD-L1 to evade antitumor immunity through interaction with programmed cell death protein 1 (PD-1) on T cells. Recent whole-genome sequence studies revealed frequent gene amplification of PD-L1 in Epstein–Barr virus-associated gastric cancer (EBVaGC). To investigate the significance of PD-L1 in cancer cells and their microenvironment in EBVaGC, we studied PD-L1 expression by analysis of the public database and immunohistochemistry with fluorescent in situ hybridization (FISH) of the PD-L1 gene. Analysis of the database from The Cancer Genome Atlas also disclosed high expression of PD-L1 in EBVaGC compared with other molecular subtypes of GC. Expression of PD-L1 was frequently detected in cancer cells of EBVaGC (33/96; 34%), with infiltration of PD-L1+immune cells in its stroma (43/96; 45%). Both expression of PD-L1 in cancer cells and PD-L1+immune cell infiltration in EBVaGC were significantly correlated with diffuse histology according to Lauren’s classification and tumor invasion (pT1b or more). As a prognostic indicator, PD-L1 expression in cancer cells correlated with poor outcomes in both overall survival and disease-specific survival (P=0.0498, 0.007). PD-L1-positive cancers had dense infiltration of PD-L1+immune cells as well as CD8+and PD-1+cells in EBVaGC. FISH analysis of representative samples of the tumor demonstrated gene amplification of PD-L1 in 11% of cases. PD-L1-amplified cells corresponded to PD-L1-positive cells showing high-intensity immunohistochemical staining among cancer cells showing weak or moderate intensities. Taken together, PD-L1 expression in cancer cells and their microenvironment may contribute to the progression of EBVaGC, and gene amplification occurs as clonal evolution during progression. This specific subtype of GC infected with EBV is potentially a good candidate for immunotherapy targeting of the PD-L1/PD-1 axis.

Colorectal cancer resembling submucosal tumor (SMT) is very rare. We herein report two cases of small colon carcinoma resembling SMT (80-year-old female and 67-year-old male), which massively invaded into the submucosal layer and accompanied marked lymphatic invasion and lymph node metastasis. We also reviewed the reported cases of colorectal carcinoma resembling SMT (SMT-like group, n=70) and analyzed the clinicopathological characteristics of this group compared with typical colorectal carcinoma cases operated at our institution (control group, n=1723). Tumors in the SMT-like group were significantly smaller in size compared with the control group; the median diameter measured 22mm vs. 37mm (P<0.01), respectively. Histologically, although the tumors in the SMT-like group were small in diameter, they almost all invaded into the submucosal (T1) or deeper layer (T2–4), and the rate of poorly differentiated adenocarcinoma or mucinous adenocarcinoma was significantly higher than that in the control group (48.6% vs. 7.7%; P<0.01). In the subgroup analysis of T1 tumors, the rate of lymphatic invasion in the SMT-like group was also significantly higher than that in the control group (43.8% vs. 15.4%; P<0.01). Carcinoma resembling SMT appears to be invasive and has a high risk of lymphatic invasion even if small in size. Therefore, surgical treatment with dissection of the regional lymph nodes might be necessary in cases with any signs of massive submucosal invasion.

This is a case report of a 75-year-old man scheduled for apical resection and cystectomy of odontogenic cysts involving both maxillary central incisors who presented with a previously unknown laryngeal mass that was discovered prior to intubation. Following induction and easy mask ventilation, direct laryngoscopy revealed a large mass on the right side of the glottis that impeded passage of a standard oral endotracheal tube. Successful atraumatic intubation was performed with the combination of a video laryngoscope (King Vision, Ambu Inc, Ballerup, Denmark) and a gum elastic bougie (GEB). Although a GEB may not be used routinely for tracheal intubation, it facilitated smooth advancement of the endotracheal tube without damaging the laryngeal mass when used in combination with video laryngoscopy.

This is a case report of a 21-year-old male patient with Cornelia de Lange syndrome (CdL) and unrepaired tetralogy of Fallot scheduled for dental treatment under general anesthesia. Anticipated dental care consisted of restorative treatment and extractions. Surgical correction of the patient's congenital cardiac abnormalities had not occurred by the time of dental treatment. As such, the developed anesthetic plan included the following goals: prevention of any anoxic episodes or spell and preparation for difficult airway management due to micrognathia secondary to CdL. To help ensure adequacy of oxygenation during induction, the normal anesthetic face mask was specially modified with a hole to permit use of the fiberoptic scope during induction and intubation. With preoperative consultation involving a cardiologist as well as the use of the modified mask, general anesthesia was safely administered without any complications.

AimsHidradenoma papilliferum (HP) is a benign vulvar neoplasm that arises from anogenital mammary-like glands, and its morphology is similar to mammary intraductal papilloma. The aim of this study was to investigate oncogenic mutations involved in the tumourigenesis of HP. We focused specifically on PIK3CAand AKT1mutations, which are both reported to be detected in 33% of mammary intraductal papillomas.MethodsIn total, seven HP cases were analysed. Clinicopathological analyses and immunohistochemistry for oestrogen receptor, p63, smooth muscle actin (SMA), p53 and β-catenin were performed. Furthermore, PIK3CA, AKT1, BRAFand KRAShot spot mutations were examined by Sanger sequencing.ResultsMorphologically, all HPs had a papillary and tubular architecture with a biphasic pattern of epithelial and myoepithelial cells. Immunohistochemistry revealed that oestrogen receptor expression was restricted to epithelial cells, whereas p63 and SMA were exclusively expressed in myoepithelial cells. The patterns of p53 and β-catenin immunostaining suggested wild-type genotypes. Direct sequencing revealed the presence of somatic PIK3CAmutations (Ex9. c.1633G>A, p.E545K and Ex20. c.3140A>G, p.H1047R) in two of the HPs and an AKT1(c.49G>A, p.E17K) mutation in one. BRAFand KRASmutations were not found in any of the HP cases.ConclusionsPIK3CAand AKT1are frequently mutated in HP tumours (29% and 14%, respectively). PIK3CA/AKT1 pathway alterations in HP further support the hypothesis that HP is the vulvar (anogenital mammary-like gland) analogue of breast intraductal papilloma.

ABSTRACTThe search for an enhanced glass medium to immobilize the sludge at the Fukushima Daiichi Nuclear Power Plant is our main purpose. The iron phosphate glass (IPG) is a potential candidate as we set about assessing it by means of theoretical and experimental investigation. Based on the results of this study, the IPG showed favorable characteristics as a vitrification medium for the sludge.

Trastuzumab-based chemotherapy is now a standard approach for patients with human epidermal growth factor receptor 2 (HER2)-positive advanced gastric cancer. However, histopathologic changes after treatment have yet to be elucidated. This study aims to characterize the histologic response of gastric cancer to trastuzumab treatment and its correlation with HER2 status. Twenty-one advanced HER2-positive gastric cancers treated with trastuzumab-based chemotherapy, including 10 surgically resected specimens and 11 biopsy samples from patients with inoperable tumors, were evaluated for the histologic responses and HER2 status of residual cells. We also reviewed, as controls, 10 cases undergoing surgical resection of tumors after chemotherapy without trastuzumab. Complete and partial histologic responses were obtained in 2 and 8 of the surgical cases, respectively. HER2-positive neoplastic cells were recognized at least focally in the 8 cases. Notably, the proportion of HER2-positive cells was always higher in superficial (mucosal/submucosal) layers than in deeper layers. Three specimens contained HER2-positive neoplastic cells exclusively in the superficial area or intravascular space, whereas deeply invasive or metastatic components almost completely disappeared or were HER2 negative when still present. In contrast, HER2-negative cells or residual tumor cells in control cases tended to survive as well or better in deeply invasive areas or in metastases than in superficial areas. Biopsy samples from nonoperative patients remained HER2 positive after treatment in 8 of 11 patients. Our observations suggest that HER2-positive neoplastic cells tend to survive within superficial areas or intravascular spaces after trastuzumab therapy, even when deeply invasive or metastatic lesions responded well to therapy.

Supplemental Digital Content is available in the text.Previous studies have identified 2 clinically significant morphologic subtypes of intrahepatic cholangiocarcinoma (ICC) on the basis of anatomic location and/or histologic appearances. Recognizing that these classification schemes are not always applicable practically, this study aimed to establish a novel classification system based on mucin productivity and immunophenotype and to determine the rationale of this classification by examining the clinicopathologic and genetic characteristics of the 2 subtypes defined by this method. We retrospectively investigated 102 consecutive ICC cases and classified them on the basis of mucin productivity and immunophenotype (S100P, N-cadherin, and NCAM). We found that 42 and 56 cases were classified as type 1 and type 2 ICCs, respectively, and only 4 cases were of indeterminate type. Type 1 ICC, generally characterized by mucin production and diffuse immunoreactivity to S100P, arose less frequently in chronic liver diseases and showed higher levels of serum CEA and CA 19-9 than did type 2 ICC, which generally showed little mucin production and exhibited immunoreactivity to N-cadherin and/or NCAM. Type 1 ICC was characterized by several pathologic features, including higher frequencies of perineural invasion and lymph node metastasis. Although the log-rank test demonstrated that type 1 ICC had significantly worse survival, the multivariate Cox regression analysis showed no prognostic significance of this histologic subtype. Genetic analyses revealed that KRASmutation was significantly more frequent in type 1 ICC, whereas IDHmutation and FGFR2translocation were restricted to type 2 ICC. In conclusion, the present classification of ICC based on mucin productivity and immunophenotype identified 2 subtypes with clinicopathologic significance.

Pulmonary capillary leakage followed by influx of blood fluid into the air space of lung alveoli is a crucial step in the progression of acute lung injury (ALI). This influx is due to increased permeability of the alveolar–capillary barrier. The extracellular matrix (ECM) between the capillary and the epithelium would be expected to be involved in prevention of the influx; however, the role of the ECM remains to be addressed. Here, we show that the ECM architecture organized by periostin, a matricellular protein, plays a pivotal role in the survival of bleomycin-exposed mice. Periostin was localized in the alveolar walls. Although periostin-null mice displayed no significant difference in lung histology and air–blood permeability, they exhibited early lethality in a model of bleomycin-induced lung injury, compared with their wild-type counterparts. This early lethality may have been due to increased pulmonary leakage of blood fluid into the air space in the bleomycin-exposed periostin-null mice. These results suggest that periostin in the ECM architecture prevents pulmonary leakage of blood fluid, thus increasing the survival rate in mice with ALI. Thus, this study provides an evidence for the protective role of the ECM architecture in the lung alveoli.