Signet Ring Cell Differentiation in Salivary Duct Carcinoma with Rhabdoid Features: Report of Three Cases and Literature Review.

Salivary duct carcinoma with rhabdoid features (SDCRF) is a rare salivary tumor with poor prognosis and is proposed as a salivary counterpart of pleomorphic lobular carcinoma of the breast (PLCB). Here, we report three cases of SDC with rhabdoid features (SDCRF) mimicking PLCB. Pleomorphic adenoma (PA) component was accompanied in all the cases confirming carcinoma ex PA. One patient had frequent rhabdoid features and showed invasive growth into the surrounding tissue. The other two patients had intracapsular tumor but with rhabdoid features. The patients with intracapsular SDCRF survived for > 5 years after surgery with no evidence of recurrence, whereas the patient with extracapsular SDCRF died 10 months after biopsy, and autopsy revealed disseminated metastasis to the central nervous system. Histologically, tumor cells in all three cases resembled PLCB, with a discohesive appearance, abundant cytoplasm, enlarged hyperchromatic nuclei, and similar immunohistochemical profiles, namely loss of membranous E-cadherin, obscured expression of membranous ß-catenin, diffuse positivity of androgen receptor, gross cystic disease fluid protein-15, mitochondrial adenosine triphosphate synthase subunit ß, MUC1, and INI-1. Estrogen and progesterone receptors were negative, and HER2 immunoreactivities were variable. The tumor cells of extracapsular invasive SDCRF exhibited higher MIB-1 labeling index and more frequent intracytoplasmic lumina than those of intracapsular SDCRF. Ultrastructurally, rhabdoid cells contained intracytoplasmic lumina with microvillous structure, analogous to those reported in PLCB. No intracytoplasmic intermediate filament aggregation was observed. These observations indicate that SDCRF is a salivary counterpart of PLCB and under signet ring cell differentiation.

Antisense RNA transcripts in the blood may be novel diagnostic markers for colorectal cancer.

Numerous genetic studies have been conducted regarding the occurrence of colorectal cancer (CRC) and the prognosis using microarrays. However, adequate investigations into the diagnostic application of microarrays have yet to be performed. The simplicity and accuracy of diagnosis and prognosis tracking are important requirements for its processes, and the use of blood cells for diagnosis is considered to be suitable to meet these requirements. The patients involved in the study were 28 preoperative patients with CRC and 6 healthy individuals who served as controls. RNA was extracted from the blood cells of the patients and analyzed using a sense/antisense RNA custom microarray. In the patients with CRC, the expression levels of 20 sense RNA and 20 antisense RNA species were identified as being significantly altered compared with that of the healthy volunteers (P<0.05; fold-change, >2.0). Cluster analysis of these RNA species revealed that the top 10 antisense RNAs significantly clustered patients with cancer and healthy individuals separately. Patients with stage I or II CRC exhibited significant changes in the expression levels of 33 sense and 39 antisense RNA species, as compared with healthy volunteers (P<0.01; fold-change >2.0). Cluster analysis demonstrated that patients with stage I or II CRC and healthy volunteers formed separate clusters only among the top 20 antisense RNA species. A tracking study of expression levels of haloacid dehalogenase-like hydrolase domain-containing 1 (HDHD1) antisense RNA was performed and a significant difference was identified between the CRC and healthy groups revealing that the levels at one week and three months following surgical removal of the cancerous tissue, decreased to almost same levels of the healthy individuals. The results of the current study indicate that HDHD1 antisense RNA may serve as a potential biomarker for the prognosis of CRC.

Loss of NDRG2 Expression Confers Oral Squamous Cell Carcinoma with Enhanced Metastatic Potential.

Loss of the tumor suppressor NDRG2 has been implicated in the development of oral squamous cell carcinoma (OSCC), acting by modulating PI3K/AKT-mediated dephosphorylation of PTEN at S380/S382/T383 (STT). Here, we show that the majority of OSCC tumors with lymph node metastasis, a major prognostic factor, exhibit high levels of phosphorylated AKT-S473 and PTEN-STT and low levels of NDRG2 expression. In Ndrg2-deficient mice, which develop a wide range of tumors, we developed a model of OSCC by treatment with the tobacco surrogate 4-nitroquinoline-1-oxide (4-NQO). In this model, both the number and size of OSCC tumors were increased significantly by Ndrg2 deficiency, which also increased invasion of cervical lymph nodes. 4-NQO treatment of human OSCC cell lines exhibiting low NDRG2 expression induced epithelial-mesenchymal transition via activation of NF-κB signaling. Conversely, ectopic expression of NDRG2 reversed the EMT phenotype and inhibited NF-κB signaling via suppression of PTEN-STT and AKT-S473 phosphorylation. Our results show how NDRG2 expression serves as a critical determinant of the invasive and metastatic capacity of OSCC. Cancer Res; 77(9); 2363-74. ©2017 AACR.

Three-dimensional reconstruction of vascular arrangement including the hepatic artery and left gastric vein during gastric surgery.

BACKGROUND: During gastric surgery, precise recognition of the anatomical variations and relationships among gastric tumors and vessels, including the hepatic artery (HA) and left gastric vein (LGV), is required. We utilized a three-dimensional (3D) reconstructed image as a preoperative simulation for gastric surgery. METHODS: We retrospectively analyzed 84 patients who underwent gastrectomy at Tsukuba Medical Center Hospital. This cohort was sequentially divided into a without-3D group (n = 42) and with-3D group (n = 42), and the perioperative outcomes were compared. The 3D image could be used to classify the HA or LGV arrangement pattern. RESULTS: Regarding the HA arrangement, the right HA of 1 patient (2.3 %) was arising from the superior mesenteric artery, the left HA of 8 patients (19 %) was arising from the left gastric artery, 29 patients (69 %) presented a normal rearrangement, and 4 patients (9.5 %) exhibited other arrangements. The analysis of the LGV arrangement revealed that the LGV in 15 patients (36 %) was located on the dorsal side of the common HA, the LGV in 5 patients (12 %) was located on the ventral side of the common HA, the LGV in 12 patients (29 %) was found on the ventral side of the splenic artery, the LGV in 6 patients (14 %) was located on the dorsal side of the splenic artery, and 4 patients (9.5 %) presented other arrangements. The intraoperative blood loss in the without-3D and with-3D groups was 276 ± 430 and 157 ± 170 g, respectively (p = 0.027). CONCLUSIONS: The 3D reconstruction technique was useful for understanding and sharing anatomic information during gastric surgery.

Nasogastric tube syndrome induced by an indwelling long intestinal tube.

The nasogastric tube (NGT) has become a frequently used device to alleviate gastrointestinal symptoms. Nasogastric tube syndrome (NTS) is an uncommon but potentially life-threatening complication of an indwelling NGT. NTS is characterized by acute upper airway obstruction due to bilateral vocal cord paralysis. We report a case of a 76-year-old man with NTS, induced by an indwelling long intestinal tube. He was admitted to our hospital for treatment of sigmoid colon cancer. He underwent sigmoidectomy to release a bowel obstruction, and had a long intestinal tube inserted to decompress the intestinal tract. He presented acute dyspnea following prolonged intestinal intubation, and bronchoscopy showed bilateral vocal cord paralysis. The NGT was removed immediately, and tracheotomy was performed. The patient was finally discharged in a fully recovered state. NTS be considered in patients complaining of acute upper airway obstruction, not only with a NGT inserted but also with a long intestinal tube.

Three-dimensional reconstruction of the vascular arrangement including the inferior mesenteric artery and left colic artery in laparoscope-assisted colorectal surgery.

BACKGROUND: We performed three-dimensional (3D) reconstruction to investigate the vascular anatomy, including the inferior mesenteric artery (IMA), left colic artery (LCA), and inferior mesenteric vein (IMV), for laparoscope-assisted left-side colorectal surgery. Furthermore, we also examined the distances from the root of the IMA to the bifurcation of the LCA and to the IMV using 3D imaging. METHODS: We retrospectively analyzed 46 patients who underwent laparoscope-assisted left-side colorectal surgery via 3D surgical reconstruction at Tsukuba Medical Center Hospital. The branching patterns among the IMA, LCA, and sigmoidal colic artery (SCA) in colon cancer could be classified into three groups (types A, B, and C): type A, in which both arteries (LCA and SCA) branch off from the same point of the IMA; type B, in which the common trunk of the LCA and SCA branches off from the IMA; and type C, in which the LCA and SCA branch off separately from the IMA. The shortest length from the root of the IMA to bifurcation of the LCA and SCA branches (D mm) or to the IMV (d mm) was measured by 3D imaging. RESULTS: The mean D mm and d mm for all cases were 39.4 ± 11.2 and 27.9 ± 9.21 mm, respectively. The D mm from the IMA root to the LCA or SCA branch in types A, B, and C was 37.8 ± 9.21, 40.5 ± 12.7, and 38.6 ± 10.2 mm, respectively. Similarly, the d mm from the IMA root to the IMV in types A, B, and C was 30.2 ± 11.3, 29.9 ± 7.27, and 25.2 ± 10.3 mm, respectively. CONCLUSION: The present 3D reconstruction technique was useful for determining the 3D vascular anatomical pattern including the relative positions of the IMA, SCA, and IMV during laparoscope-assisted left-side colorectal surgery.

[Suspected dihydropyrimidine dehydrogenase deficiency in a patient receiving capecitabine as adjuvant chemotherapy after colon resection].

We report here the case of a 75-year-old male patient who developed severe side effects after treatment with capecitabine (Xeloda®) that he received as adjuvant chemotherapy. He was suspected to have partial dihydropyrimidine dehydrogenase (DPD) deficiency. The patient underwent sigmoidectomy for sigmoid cancer and was treated with capecitabine as adjuvant chemotherapy. He was admitted to our hospital 14 days after the start of treatment with appetite loss, diarrhea, and a high body temperature. After admission, he developed severe neurotoxicity (Grade 4). We measured the DPD activity in peripheral mononuclear cells, which indicated partial DPD deficiency.

Development of a complete human anti-human transferrin receptor C antibody as a novel marker of oral dysplasia and oral cancer.

Oral squamous cell carcinoma (OSCC) is the sixth most common cancer worldwide. Up to 20% of oral dysplasia cases have been suggested to undergo malignant transformation to OSCC; however, there are no methods to predict OSCC development. In this study, to identify the genes associated with oral dysplasia progression, we performed genomic copy number analyses of genomic DNA samples isolated from primary oral dysplasia and OSCC via the microdissection method and found elevated expression of transferrin receptor C (TfR1/TFRC) with genomic amplification in oral dysplasia and OSCC. The expression rate of TFRC in OSCC was significantly higher than that in dysplasia, suggesting that OSCC disease progression might be related to TFRC expression. Additionally, we investigated the in vitro and in vivo impacts of a newly established anti-human TFRC monoclonal antibody, which was isolated from a human cDNA library using the phage-display method, on cell proliferation and survival. The anti-TFRC antibody blocked the interaction between transferrin and TFRC and consequently inhibited iron uptake, leading to the iron deprivation-mediated suppression of cell growth and induction of apoptosis. Moreover, we demonstrated that the anti-TFRC antibody efficiently inhibited tumor growth in a murine xenograft OSCC model. Therefore, we suggest our developed complete human anti-human TFRC antibody as a useful, novel treatment for oral dysplasia and OSCC.

Sparse phase-stepping in two-dimensional x-ray phase contrast imaging.

We have developed a sparse phase-stepping (SPS) method for x-ray Talbot-Lau interferometry, which first constructs a SPS intensity pattern of fewer images than the conventional phase-stepping (PS) method and then fills the data gap with neighboring pixels for phase retrieval. The SPS method is highly beneficial in practice since the fundamental difference in spatial resolution between the SPS and PS methods becomes negligible due to the blur caused by an interferometer. The concept of the SPS method has been proved by the experiment using a small effective source size. Furthermore, the experiment using a large effective source size has verified that in practical situations the SPS method can reduce the required number of images for phase retrieval and still offer the retrieved images with as high a spatial resolution as the PS method.

A phase demodulation method with high spatial resolution for two-dimensional single-shot X-ray Talbot interferometry.

A new phase demodulation approach is proposed that uses windowed Fourier transforms to achieve high spatial resolution in fringe pattern analysis with a high signal-to-noise ratio for single-shot X-ray grating-based interferometry. Conventionally, Fourier transforms have been used to demodulate single-fringe patterns, but this requires a fringe pattern with a long period to obtain an acceptable signal-to-noise ratio among the demodulated parameters. However, by controlling the signal-to-noise ratio, the spatial resolution of demodulated parameters is degraded below that obtained from the phase-stepping method, which requires several images to obtain these parameters. In this paper, we introduce the use of a windowed Fourier transform with a process for analyzing the objective spectrum in isolation from other spectra on the Fourier space to overcome the limitations of the Fourier transform method. It is proved that with suitable assumptions the objective spectrum is isolated theoretically, and the spatial resolution is improved by practically accepting the limitations from the assumptions. We demonstrate the validity of the proposed method by comparing the modulation transfer function of a synthetic phantom with the conventional FT method. The proposed method is also valid on practical data obtained by an experimental setup, by which it is demonstrated that a high spatial resolution with high signal-to-noise ratio can be achieved by our proposed method.

Loss of NDRG2 expression activates PI3K-AKT signalling via PTEN phosphorylation in ATLL and other cancers.

Constitutive phosphatidylinositol 3-kinase (PI3K)-AKT activation has a causal role in adult T-cell leukaemia-lymphoma (ATLL) and other cancers. ATLL cells do not harbour genetic alterations in PTEN and PI3KCA but express high levels of PTEN that is highly phosphorylated at its C-terminal tail. Here we report a mechanism for the N-myc downstream-regulated gene 2 (NDRG2)-dependent regulation of PTEN phosphatase activity via the dephosphorylation of PTEN at the Ser380, Thr382 and Thr383 cluster within the C-terminal tail. We show that NDRG2 is a PTEN-binding protein that recruits protein phosphatase 2A (PP2A) to PTEN. The expression of NDRG2 is frequently downregulated in ATLL, resulting in enhanced phosphorylation of PTEN at the Ser380/Thr382/Thr383 cluster and enhanced activation of the PI3K-AKT pathway. Given the high incidence of T-cell lymphoma and other cancers in NDRG2-deficient mice, PI3K-AKT activation via enhanced PTEN phosphorylation may be critical for the development of cancer.

A phase demodulation method for two-dimensional grating-based X-ray interferometry.

This paper presents a novel approach to achieving high spatial resolution in the demodulation of images produced by a two-dimensional X-ray Talbot interferometry (XTI) system. Currently, demodulation of XTI images is mainly performed by either phase-stepping (PS) or Fourier transform (FT) methods. However, the PS method for two-dimensional XTI demodulation requires a larger number of exposures and a more complex grating control process than that of one-dimensional XTI. On the other hand, although the FT method uses only a single-fringe image, it gives lower spatial resolution than the PS method. For practical application of two-dimensional XTI, a simpler exposure process with high spatial resolution is required. In this paper, we introduce a hybrid method combining the PS and FT methods. This method simplifies the exposure process in comparison with the PS method required in two-dimensional XTI while achieving higher spatial resolution than the FT method in the demodulation of images. The method works by using additional exposures to eliminate unnecessary spectral components that appear in the FT method. Furthermore, the proposed method is demonstrated by using actual two-dimensional XTI data and shown to achieve high spatial resolution in the demodulation of images for both the x- and y-differential phase components.

Differential expression profiles of sense and antisense transcripts between HCV-associated hepatocellular carcinoma and corresponding non-cancerous liver tissue.

Recent studies have demonstrated that natural antisense transcripts, which are complementary sequences to messenger RNA, have important cellular functions such as the stabilization and silencing of mRNA. However, the possible contribution of antisense transcripts in hepatocellular carcinoma (HCC) development has not been described. Therefore, we simultaneously investigated the sense and antisense transcripts of HCC and non-cancerous tissues to explore the possible contribution of antisense transcripts to HCC progression. RNA was prepared from 15 HCV-associated HCCs and from 6 corresponding non-cancerous tissues and was subjected to expression profile analysis of sense and antisense transcripts using a human custom microarray. Differential expression of 161 sense and 25 antisense transcripts was observed with more than 2-fold between HCC and non-cancerous tissue (p<0.001). The expression of the sense and antisense transcripts was used to cluster cancer and non-cancerous tissues, and the cancer and non-cancerous tissues were found to be clearly separated into different clusters. Additionally, the sense and antisense expression profiles were analyzed with regard to HCC differentiation (p<0.001), resulting in 71 sense and 43 antisense transcripts. These unique transcripts did not overlap with those found in the discrimination of HCC from non-cancerous tissues. When the HCC tissues were clustered by transcript expression, the antisense transcripts resulted in clustering of HCC that was consistent with grouping based on histology. These findings strongly indicate that the antisense transcripts together with the sense transcripts are involved in liver tumorigenesis.

Overexpression of the DNA sensor proteins, absent in melanoma 2 and interferon-inducible 16, contributes to tumorigenesis of oral squamous cell carcinoma with p53 inactivation.

The development of oral squamous cell carcinoma (OSCC) is a multistep process that requires the accumulation of genetic alterations. To identify genes responsible for OSCC development, we performed high-density single nucleotide polymorphism array analysis and genome-wide gene expression profiling on OSCC tumors. These analyses indicated that the absent in melanoma 2 (AIM2) gene and the interferon-inducible gene 16 (IFI16) mapped to the hematopoietic interferon-inducible nuclear proteins. The 200-amino-acid repeat gene cluster in the amplified region of chromosome 1q23 is overexpressed in OSCC. Both AIM2 and IFI16 are cytoplasmic double-stranded DNA sensors for innate immunity and act as tumor suppressors in several human cancers. Knockdown of AIM2 or IFI16 in OSCC cells results in the suppression of cell growth and apoptosis, accompanied by the downregulation of nuclear factor kappa-light-chain-enhancer of activated B cells activation. Because all OSCC cell lines have reduced p53 activity, wild-type p53 was introduced in p53-deficient OSCC cells. The expression of wild-type p53 suppressed cell growth and induced apoptosis via suppression of nuclear factor kappa-light-chain-enhancer of activated B cells activity. Finally, the co-expression of AIM2 and IFI16 significantly enhanced cell growth in p53-deficient cells; in contrast, the expression of AIM2 and/or IFI16 in cells bearing wild-type p53 suppressed cell growth. Moreover, AIM2 and IFI16 synergistically enhanced nuclear factor kappa-light-chain-enhancer of activated B cells signaling in p53-deficient cells. Thus, expression of AIM2 and IFI16 may have oncogenic activities in the OSCC cells that have inactivated the p53 system.

Two-dimensional gratings-based phase-contrast imaging using a conventional x-ray tube.

A Talbot-Lau interferometer using two-dimensional gratings and a conventional x-ray tube has been used to investigate a phase-contrast imaging technique that is sensitive to phase gradients in two orthogonal directions. Fourier analysis of Moiré fringe patterns was introduced to obtain differential phase images and scattering images from a single exposure. Two-dimensional structures of plastic phantoms and characteristic features of soft tissue were clearly obtained at 17.5 keV. The phase-stepping technique was also examined to investigate the spatial resolution of different phase retrieval methods. In the presented setup we found that the choice of phase retrieval method made little difference in image blur, and a large effective source size was found to give a high intensity in the image plane.

Two-dimensional grating-based X-ray phase-contrast imaging using Fourier transform phase retrieval.

We demonstrate a single shot two-dimensional grating-based X-ray phase-contrast imaging using a synchrotron radiation source. A checkerboard designed phase grating for π phase modulation at 17 keV and 35 keV, and a lattice-shaped amplitude grating with a high aspect ratio to shield X-rays up to 35 keV were fabricated. A Fourier analysis of Moiré fringe generated by the gratings was introduced to obtain the two-dimensional differential phase-contrast image with a single exposure. The results show that soft tissues and cartilages of a chicken wing sample are clearly seen with differential phase variation in two-dimensional directions. Using this method not only the whole of an object but also only an inner part of the object can be imaged.

Identification of natural antisense transcripts involved in human colorectal cancer development.

Natural antisense transcripts (NATs) constitute a class of non-coding RNAs that have emerged as important regulators of gene expression. However, involvement of NATs in colorectal cancer (CRC) development has not been reported to date. In the present study, the up- and down-regulation of NATs were investigated in human CRC for their possible involvement in CRC development. Total RNAs isolated from 51 CRC tissues, 9 corresponding non-cancerous tissues and 19 liver metastatic tissues from surgically resected samples were subjected to expression analysis using a custom-microarray containing human sense/antisense probes for ca. 21,000 genes. Comparing CRC tissues with non-cancerous tissues, we identified 415 NATs differentially expressed in CRC and non-cancerous tissues to a significant degree (p<0.001, fold change >4.0 or ≤4.0). When a hierarchical clustering was performed on CRC and non-cancerous samples using these 415 NATs, the samples were separately clustered. Principal component analysis with the same NATs showed clear separation of CRC and non-cancerous samples using the first two principal components (PC1, 80%; PC2, 10%). To validate the expression results obtained from the microarray, the expressions of the 3 selected NATs were examined by strand-specific RT-qPCR, revealing that these expression profiles were consistent with those obtained from microarray analysis. In addition, the NAT expression patterns were found to be different between primary tumors with liver metastasis and those without liver metastasis. In conclusion, these findings taken together indicated that NATs identified in the present study would be involved in CRC development as well as possibly in its metastasis.

Laparoscopy-assisted esophagoenteral anastomosis using endoscopic purse-string suture instrument "Endo-PSI (II)" and circular stapler.

It is said that laparoscopic esophagoenteral anastomosis is not easy. In particular, purse-string suture of the abdominal esophagus is difficult when using a circular stapler. We have developed an endoscopic purse-string suture instrument, the "Endo-PSI (II)", and the instrument was employed clinically during laparoscopy-assisted total gastrectomy. The device was inserted into the abdominal cavity through a 4-cm minilaparotomy of the epigastrium, and pneumoperitoneum was established by closing a Lap Disc. The Endo-PSI (II) was attached to the abdominal esophagus and a straight needle with a 2-0 polypropylene suture was passed through the device laparoscopically. After a purse-string suture of the abdominal esophagus was made, the abdominal esophagus was transected laparoscopically and the removed stomach was pulled out through the minilaparotomy. The anvil head of a circular stapler was inserted into the abdominal cavity through the minilaparotomy, and insertion of the anvil into the esophagus and ligation of the purse-string suture were performed laparoscopically, too. The combination of using a circular stapler for esophagojejunostomy and closure of the jejunal stump was also performed laparoscopically. Between May 2007 and May 2008, these products were used in 23 patients during laparoscopy-assisted total gastrectomy. There were no cases that required conversion to a conventional open procedure. The newly developed Endo-PSI (II) was useful for laparoscopic purse-string suture of the esophagus.

A novel cinnamic acid derivative that inhibits Cdc25 dual-specificity phosphatase activity.

The Cdc25 dual-specificity phosphatases are key regulators of cell cycle progression through activation of cyclin-dependent kinases (Cdk). Three homologs exist in humans: Cdc25A, Cdc25B, and Cdc25C. Cdc25A and Cdc25B have oncogenic properties and are overexpressed in some types of tumors. Compounds that inhibit Cdc25 dual-specificity phosphatase activity might thus be potent anticancer agents. We screened several hundred compounds in a library using an in vitro phosphatase assay, with colorimetric measurement of the conversion of p-nitrophenyl phosphate (pNPP) to p-nitrophenol by the catalytic domain of recombinant human Cdc25, and discovered TPY-835, which inhibits Cdc25A and Cdc25B activity (IC50 = 5.1 and 5.7 microM, respectively). TPY-835 had mixed inhibition kinetics for Cdc25A and Cdc25B. TPY-835 caused cell cycle arrest in the G1 phase in human lung cancer cells (A549 and SBC-5) but not cell cycle arrest in the G2/M phase. After treatment with TPY-835, the activation of Cdk2 was suppressed and phosphorylation of the retinoblastoma (Rb) protein was decreased in SBC-5 cells. In addition, TPY-835 induced an increase of the sub-G1 phase cell population after 48-72 h treatment. The growth inhibitory effects of TPY-835 against cisplatin (CDDP)-, camptothecin- and 5-FU-resistant cell lines are comparable to the growth inhibitory effect on their parental lines, thus indicating that TPY-835 did not show cross-resistance to these cell lines. These results suggest that TPY-835 is a promising candidate for constructing a novel class of antitumor agents that can control the cell cycle progression of cancer cells.