[CA19-9 Producing Gastric Cancer with Early Recurrence of Multiple Colon Metastases Following Gastrectomy-A Case Report].

A 90-year-old male underwent total gastrectomy for gastric cancer 10 months earlier. The cancer was pathologically diagnosed as Stage ⅢA. Preoperative serum CA19-9 level was as high as 1,326 U/mL, but quickly decreased after surgery. Although the serum CA19-9 level gradually re-increased, CT did not reveal recurrence of the disease. Ten months following surgery, the patient visited our hospital due to vomiting, and ileus was suspected because of finding of sigmoid colon tumors in the abdominalCT. Colonoscopy showed a circumferentialtumor with severe stenosis in the sigmoid colon, which was diagnosed as tubular adenocarcinoma by biopsy. After preoperative diagnosis of multiple colon cancers, sigmoidectomy was performed. A total of 4 tumors were revealed in the resected specimen. Pathological findings showed cancer cells with nuclear atypia in all tumors, which was very similar to findings in the previous gastric cancer. Immunohistochemical staining confirmed high expression of CA19-9 in both gastric and colon tumors. We concluded that the tumors were metastases of the CA19-9 producing gastric cancer.

The morphogenesis-related NDR kinase pathway of Colletotrichum orbiculare is required for translating plant surface signals into infection-related morphogenesis and pathogenesis.

Plant infection by pathogenic fungi involves the differentiation of appressoria, specialized infection structures, initiated by fungal sensing and responding to plant surface signals. How plant fungal pathogens control infection-related morphogenesis in response to plant-derived signals has been unclear. Here we showed that the morphogenesis-related NDR kinase pathway (MOR) of the cucumber anthracnose fungus Colletotrichum orbiculare is crucial for appressorium development following perception of plant-derived signals. By screening of random insertional mutants, we identified that the MOR element CoPag1 (Perish-in-the-absence-of-GYP1) is a key component of the plant-derived signaling pathway involved in appressorium morphogenesis. Constitutive activation of the NDR kinase CoCbk1 (Cell-wall-biosynthesis-kinase-1) complemented copag1 defects. Furthermore, copag1 deletion impaired CoCbk1 phosphorylation, suggesting that CoPag1 functions via CoCbk1 activation. Searching for the plant signals that contribute to appressorium induction via MOR, we found that the cutin monomer n-octadecanal, degraded from the host cuticle by conidial esterases, functions as a signal molecule for appressorium development. Genome-wide transcriptional profiling during appressorium development revealed that MOR is responsible for the expression of a subset of the plant-signal-induced genes with potential roles in pathogenicity. Thus, MOR of C. orbiculare has crucial roles in regulating appressorium development and pathogenesis by communicating with plant-derived signals.

[Long-Term Survival in a Patient with Combined Hepatocellular-Cholangiocarcinoma Treated with Multidisciplinary Therapy - A Case Report].

The patient was a woman in her 50's. She went to the hospitalfor epigastric discomfort. Numerous hypovascular tumors spreading almost to the entire liver were detected via abdominal CT. We diagnosed the tumors as unresectable intrahepatic cholangiocarcinoma(ICC). Chemotherapy with gemcitabine(GEM)and S-1 was given. The tumors shrunk markedly, and stable disease status was maintained for 2 years and 4 months. Thereafter, the regimen was changed to GEM plus cisplatin (CDDP)because of progressive disease, which was then uncontrolled. Although proton beam therapy and radiofrequency ablation were subsequently performed, multiple lesions appeared in the liver, and metastasis was also observed shortly in the left lung. Right hepatic trisegmentectomy and thoracoscopic left lung partial resection were performed in 2 stages. Histopathological findings showed morphological hepatocellular carcinoma(HCC)as well as positive immunostaining with CEA, CK7, and CK19. The final diagnosis was combined hepatocellular-cholangiocarcinoma. The patient is without recurrence for 9 months following operation, and long-term survivalfor 4 years and 6 months has been obtained.

Observational cohort study focused on treatment continuity of patients administered XELOX plus bevacizumab for previously untreated metastatic colorectal cancer.

BACKGROUND: There has been remarkable progress in systemic chemotherapy for metastatic colorectal cancer due to the widespread use of irinotecan, oxaliplatin, anti-vascular endothelial growth factor antibody, and anti-epidermal growth factor receptor antibody. It is important to continue treatment with the optimal combination of these drugs and prolong progression-free survival (PFS) to improve overall survival (OS). We conducted a prospective observational cohort study of 40 patients treated with XELOX plus bevacizumab for previously untreated metastatic colorectal cancer to investigate treatment continuity. PATIENTS AND METHODS: Eligibility criteria were as follows: 1) histologically confirmed metastatic colorectal cancer; 2) lesions evaluable by imaging; 3) previously untreated; 4) suitable condition to receive XELOX plus bevacizumab; and 5) written informed consent. Outcomes were treatment continuity, overall response rate, resection rate, liver resection rate, time to treatment failure, PFS, and OS. Forty patients were enrolled and followed up for 2 years. RESULTS: Between July 2010 and June 2012, 40 patients were enrolled. The median number of treatment cycles was 7.5, and the reasons for discontinuation of treatment were as follows: complete response (five patients), resection (ten patients), progression (15 patients), adverse events (seven patients), and patient refusal (three patients). The overall response rate was 57.5%, resection rate was 25%, and liver resection rate was 15%. After a median follow-up of 31.4 months, the median time to treatment failure, PFS, and OS were 5.3, 13.3, and 38.9 months, respectively. CONCLUSION: Although the median time to treatment failure was 5.3 months, the median PFS and OS were prolonged to 13.3 and 38.9 months, respectively. This may have resulted from the chemotherapy-free interval due to complete response in five patients and resection in ten patients.

Effects of macroporous hydroxyapatite carriers on the growth and function of human hepatoblasts derived from fetal hepatocytes.

Improvement of three-dimensional (3D) culture conditions, including substrates for cell growth, is needed for various cell-based applications. In this study, we developed hydroxyapatite (HAp) macroporous carriers having several pore size distributions and tried to obtain the findings about the effective pore sizes for the growth and function of hepatoblasts derived from human fetal hepatocytes. Cellular CYP3A4 activity was significantly enhanced when 20% HAp macroporous carrier was used, reaching 1.49±0.28 pmol/10(6) cells/min of benzyloxyresorufin-O-dealkylation activity, which is comparable to that of primary human hepatocytes from livers of adult donors. Analysis of the pore size (the radius of curvature) distribution of each HAp carrier using a 3D-electron beam surface roughness analyzer revealed two peaks of pore size distribution at 30-40 µm and 70-80 µm, respectively. Thirty-five percent of the pores in the 20% carrier had a size distribution within 50-80 µm. Especially, pores of 70-80 µm were more abundant in the 20% HAp carrier than in the 10% and 30% HAp carriers. These results suggested that a HAp carrier with the pore size distribution of 50-80 µm might be effective for cell growth and function in human hepatoblasts derived from fetal hepatocytes.

[Multiple Endocrine Neoplasia Type 1(MEN1)Presenting with Hypoglycemic Attacks - A Case Report].

A woman in her 60s visited our hospital because of frequent hypoglycemia and episodes of unconsciousness over the last 6 years. A 4 cm tumor was detected on the pancreatic tail using abdominal computed tomography and ultrasonography. An insulinoma was strongly suspected from the results of the fasting test and glucagon load test, and a distal pancreatectomy with splenectomy was performed. Pathological examination indicated an insulinoma and neuroendocrine tumor(NET)G2 based on the WHO 2010 classification. The patient's blood sugar and insulin levels returned to normal, and hypoglycemic attacks disappeared postoperatively. Six months later, a total parathyroidectomy was performed because of primary hyperparathyroidism with hypertrophy of the parathyroid glands. Furthermore, pituitary swellingwas also detected usinghead MRI. However, the patient has been under observation because the tumor was non-functional without any associated symptoms. Thus, we diagnosed the patient with multiple endocrine neoplasia type 1(MEN1). In patients with pancreatic NET, it is necessary to consider the possibility of MEN1.

[Successful Placement of Self-Expandable Metallic Stent for Malignant Gastrojejunostomy Obstruction Following Pancreatoduodenectomy - A Case Report].

A man in his 50 s, who had undergone subtotal stomach-preserving pancreatoduodenectomy with modified Child's reconstruction for pancreatic cancer 8 months back, was hospitalized because of vomiting and difficulty in feeding.Radiological and endoscopic studies revealed a malignant obstruction of the gastrojejunostomy site due to peritoneal recurrence of the cancer.Although a self-expandable metallic stent(SEMS)was placed in the anastomotic site, it slipped back into the stomach 3 days later.It was suggested that the migration was caused by antiperistalsis, because the SEMS was placed in the afferent loop.Although the SEMS was required to be placed in concordance with the peristaltic direction, it was impossible to pass a guidewire directly from the stomach into the efferent loop.Therefore, the guidewire was placed antidromically through a narrow site from the distal portion of the efferent loop via Braun anastomosis, and the SEMS was subsequently placed without any complication.This allowed the patient to maintain oral intake throughout his remaining life.Our antidromic approach for SEMS placement could be beneficial if performing a standard procedure is difficult.

[Gastric Cancer Diagnosed with Metastasis of the Navel(Sister Mary Joseph's Nodule) - A Case Report].

A woman in her 60s was admitted to our hospital with pain and induration of the navel. She was diagnosed with gastric cancer with metastasis to the navel and underwent total gastrectomy and navel extraction. Because disseminated nodules were detected in the Douglas pouch and sigmoid colon, sigmoidectomy was performed to prevent bowel obstruction. The navel tumor was histologically diagnosed as a metastasis of the gastric cancer. One month after surgery, a chest skin tumor, which was also a skin metastasis of the gastric cancer[T4aN3M1(SKI, OTH)H0P1, fStage IV ], was detected, and tumor enucleation was performed. Enucleation was followed by 47 courses of systemic chemotherapy consisting of capecitabine, cisplatin, and trastuzumab. No recurrence or metastasis has been observed via FDG-PET/CT as of 5 years after surgery. Gastric cancer with peritoneal dissemination in addition to navel metastasis has been reported to have an extremely poor prognosis. However, long-term, recurrence-free survival was obtained in this case owing to aggressive surgical resection, followed by persistent systemic chemotherapy.

[Adjuvant Surgery for Initially Unresectable Locally Advanced Pancreatic Cancer following Gemcitabine and S-1 Chemotherapy--A Case Report].

A man in his 60s was admitted with obstructive jaundice. A hypovascular tumor, 55 mm in diameter, was detected in the pancreas head on imaging. The superior mesenteric vein showed severe stenosis bilaterally and the roots of all branches were invaded by the tumor. The tumor was diagnosed as unresectable pancreatic cancer, and chemotherapy of gemcitabine and S-1 was administered, resulting in a remarkable reduction of the tumor size. Following 7 courses of chemotherapy, a subtotal stomach-preserving pancreatoduodenectomy was carried out. Microscopic examination revealed no residual cancer cells in the resected specimen, indicating that pathological complete remission was obtained. Although some reports suggest that surgical treatment for patients with initially unresectable pancreas cancer who show excellent response to chemotherapy may improve the prognosis, further studies are needed.

[Surgical treatment for gall bladder cancer with a confluence stone-a case report].

The patient was a 71-year-old man. In September 2011, he experienced abdominal pain with high fever. Abdominal computed tomography (CT) diagnosed acute cholecystitis with a confluence stone (corlette classification type II). He underwent total cholecystectomy and placement of a T-tube in the main bile duct through the gall bladder duct. However, pathological investigations revealed gall bladder cancer in the neck and body part of the gall bladder, leading to a diagnosis of gall bladder adenocarcinoma(Gbn, Flat type, tub2, INF ß,pSS, pHinf0, pBinf1, pPV0, pA0, pT3) with a confluence stone. We suspected that the tumor was present in the common bile duct. Therefore, in October 2011, he underwent choledochectomy, resection of the liver bed, lymph node dissection, and choledocho-jejunostomy. Pathological findings revealed that the tumor was present in the common bile duct. He died 8 months after the last surgery because of recurrence of peritoneal metastasis.

Arbuscular mycorrhizal fungal spores host bacteria that affect nutrient biodynamics and biocontrol of soil-borne plant pathogens.

The aim of this research was to isolate and characterize bacteria from spores of arbuscular mycorrhizal fungi (AMF). We designated these bacteria 'probable endobacteria' (PE). Three bacterial strains were isolated from approximately 500 spores of Gigaspora margarita (Becker and Hall) using a hypodermic needle (diameter, 200 µm). The bacteria were identified by morphological methods and on the basis of ribosomal gene sequences as Bacillus sp. (KTCIGM01), Bacillus thuringiensis (KTCIGM02), and Paenibacillus rhizospherae (KTCIGM03). We evaluated the effect of these probable endobacteria on antagonistic activity to the soil-borne plant pathogens (SBPPs) Fusarium oxysporum f. sp. lactucae MAFF 744088, Rosellinia necatrix, Rhizoctonia solani MAFF 237426, and Pythium ultimum NBRC 100123. We also tested whether these probable endobacteria affected phosphorus solubilization, ethylene production, nitrogenase activity (NA), and stimulation of AMF hyphal growth. In addition, fresh samples of spores and hyphae were photographed using an in situ scanning electron microscope (SEM) (Quanta 250FEG; FEI Co., Japan). Bacterial aggregates (BAs), structures similar to biofilms, could be detected on the surface of hyphae and spores. We demonstrate that using extraction with an ultrathin needle, it is possible to isolate AMF-associated bacterial species that are likely derived from inside the fungal spores.

Cell sheet technology for regeneration of esophageal mucosa.

The progress of tissue-engineering technology has realized development of new therapies to treat various disorders by using cultured cells. Cell- and tissue-based therapies have been successfully applied to human patients, and several tissue-engineered products have been approved by the regulatory agencies and are commercially available. In the review article, we describe our experience of development and clinical application of cell sheet-based regenerative medicine. Endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD) have been shown to be useful for removal of gastrointestinal neoplasms with less invasiveness compared with open surgery, especially in esophageal surgery. However, postoperative inflammation and stenosis are major complications observed after intensive mucosal resection. Therefore, we have developed novel regenerative medicine to prevent such complications and promote wound healing of esophageal mucosa after EMR or ESD. Transplantable oral mucosal epithelial cell sheets were fabricated from patients' own oral mucosa. Immediately after EMR or ESD, fabricated autologous cell sheets were endoscopically transplanted to the ulcer sites. We performed a preclinical study with a canine model. In human clinical settings, cell culture and cell sheet fabrication were performed in clean rooms according to good manufacturing practice guidelines, and pharmaceutical drugs were used as supplements to culture medium in place of research regents used in animal study. We believe that cell-based regenerative medicine would be useful to improve quality of life of patients after EMR or ESD.

Phytochemicals to suppress Fusarium head blight in wheat-chickpea rotation.

Fusarium diseases cause major economic losses in wheat-based crop rotations. Volatile organic compounds (VOC) in wheat and rotation crops, such as chickpea, may negatively impact pathogenic Fusarium. Using the headspace GC-MS method, 16 VOC were found in greenhouse-grown wheat leaves: dimethylamine, 2-methyl-1-propanol, octanoic acid-ethyl ester, acetic acid, 2-ethyl-1-hexanol, nonanoic acid-ethyl ester, nonanol, N-ethyl-benzenamine, naphthalene, butylated hydroxytoluene, dimethoxy methane, phenol, 3-methyl-phenol, 3,4-dimethoxy-phenol, 2,4-bis (1,1-dimethyethyl)-phenol, and 1,4,7,10,13,16-hexaoxacyclooctadecane; and 10 VOC in field-grown chickpea leaves: ethanol, 1-penten-3-ol, 1-hexanol, cis-3-hexen-1-ol, trans-2-hexen-1-ol, trans-2-hexenal, 3-methyl-1-butanol, 3-hydroxy-2-butanone, 3-methyl-benzaldehyde and naphthalene. Also found was 1-penten-3-ol in chickpea roots and in the root nodules of two of the three cultivars tested. Chickpea VOC production pattern was related (P=0.023) to Ascochyta blight severity, suggesting that 1-penten-3-ol and cis-3-hexen-1-ol were induced by Ascochyta rabiei. Bioassays conducted in Petri plates established that chickpea-produced VOC used in isolation were generally more potent against Fusarium graminearum and Fusarium avenaceum than wheat-produced VOC, except for 2-ethyl-1-hexanol, which was rare in wheat and toxic to both Fusarium and tetraploid wheat. Whereas exposure to 1-penten-3-ol and 2-methyl-1-propanol could suppress radial growth by over 50% and octanoic acid-ethyl ester, nonanol, and nonanoic acid-ethyl ester had only weak effects, F. graminearum and F. avenaceum growth was completely inhibited by exposure to trans-2-hexenal, trans-2-hexen-1-ol, cis-3-hexen-1-ol, and 1-hexanol. Among these VOC, trans-2-hexenal and 1-hexanol protected wheat seedlings against F. avenaceum and F. graminearum, respectively, in a controlled condition experiment. Genetic variation in the production of 2-ethyl-1-hexanol, a potent VOC produced in low amount by wheat, suggests the possibility of selecting Fusarium resistance in wheat on the basis of leaf VOC concentration. Results also suggests that the level of Fusarium inoculum in chickpea-wheat rotation systems may be reduced by growing chickpea genotypes with high root and shoot levels of trans-2-hexen-1-ol and 1-hexanol.

Probing force-induced unfolding intermediates of a single staphylococcal nuclease molecule and the effect of ligand binding.

Single-molecule manipulation techniques have given experimental access to unfolding intermediates of proteins that are inaccessible in conventional experiments. A detailed characterization of the intermediates is a challenging problem that provides new possibilities for directly probing the energy landscape of proteins. We investigated single-molecule mechanical unfolding of a small globular protein, staphylococcal nuclease (SNase), using atomic force microscopy. The unfolding trajectories of the protein displayed sub-molecular and stochastic behavior with typical lengths corresponding to the size of the unfolded substructures. Our results support the view that the single protein unfolds along multiple pathways as suggested in recent theoretical studies. Moreover, we found the drastic change, caused by the ligand and inhibitor bindings, in the mechanical unfolding dynamics.

Synthesis of diphenylmethyl analogues and their affinity for the melanocortin-4 receptor and the serotonin transporter.

While examining antagonists of the melanocortin-4 receptor (MC4 receptor), we found that compound 12b, containing a diphenylmethyl moiety, had a relatively high affinity for the MC4 receptor. When diphenylmethyl analogues were further examined, compounds 12c and 18 were also found to exhibit a high affinity for the MC4 receptor (IC(50)=46.7 nM and 33.2 nM, respectively). Furthermore, compound 12c was also found to show a high affinity for the serotonin transporter (IC(50)=10.7 nM). Here, we describe the synthesis and biological evaluation of various diphenylmethyl analogues in relation to their actions on the MC4 receptor and the serotonin transporter.

Novel piperazines: potent melanocortin-4 receptor antagonists with anxiolytic-like activity.

In the present study, we found that a novel piperazine compound, 11a, showed a moderate affinity (IC(50)=333nM) for the MC4 receptor. We developed the new type of piperazine compounds and found that mono-piperazine 11b exhibited a high-affinity (IC(50)=40.3nM) for the MC4 receptor. We also found that a series of biphenyl analogues exhibited a high-affinity for the receptor, and in particular, compound 11j exhibited the highest affinity for the MC4 receptor with an IC(50) value of 14.5nM. Furthermore, some of these compounds, when administered orally, significantly reversed the stress-induced anxiety-like behavior in rats. In this paper, we report the synthesis, structure-activity relationships, and oral activity of the novel mono-piperazines as MC4 receptor antagonists.

Structure-activity relationships of novel piperazines as antagonists for the melanocortin-4 receptor.

During the investigation of antagonists for the MC4 receptor, we found that 10ab having a naphthyl group showed almost the same binding affinity for the MC4 receptor as that of the lead compound 1 with a benzoyl group. We also developed a new type of compounds, namely, bis-piperazines, and found that the bis-piperazines 10 exhibited a high affinity for the MC4 receptor. In particular, (-)-10bg exhibited the highest affinity for the MC4 receptor with an IC50 value of 8.13nM. In this paper, we present the design, synthesis, and structure-activity relationships of the novel bis-piperazines as MC4 receptor antagonists.

Design and synthesis of 1-(4-benzoylphenyl)imidazole derivatives as new potent 20-HETE synthase inhibitors.

Structural modification of the novel 20-HETE synthase inhibitor 1 (IC(50) 310nM) is described. Introduction of a side chain with a carboxylic acid at the terminal position to 1 resulted in increased ability to inhibit human renal microsomal production of 20-HETE (7c: IC(50) 7.9nM), with good selectivity toward CYP2D6 and cyclooxygenases (COX)-1 and -2.

Rapid genetic diagnosis with the transcription-reverse transcription concerted reaction system for cancer micrometastasis.

BACKGROUND: Detection of cancer micrometastases is required for improvement of cancer therapy. The aim of this study was to establish a rapid and practical genetic assay to detect micrometastasis in gastric cancer and to assess its clinical significance with respect to prognosis. METHODS: A novel RNA amplification system with transcription-reverse transcription concerted reaction (TRC) was introduced for quantitative detection of cancer-specific carcinoembryonic antigen messenger RNA. The sensitivity and quantitative aspects of the assay were assessed with the full-length carcinoembryonic antigen messenger RNA, a gastric cancer cell line (MKN-45), and metastatic lymph nodes obtained from patients with gastric cancer. Peritoneal lavage fluid specimens that were collected from gastric cancer surgery were subjected to the assay, and the clinical significance of the results was examined for prediction of recurrence and survival. RESULTS: The quantification, sensitivity, and reproducibility of the assay with the TRC reaction were equal to those of quantitative reverse transcriptase-polymerase chain reaction with LightCycler. The most important advantages of the assay were its simplicity and rapidity. Molecular diagnosis of peritoneal lavage fluid by the TRC reaction significantly correlated with depth of invasion, peritoneal metastasis, clinical stage, overall survival, and peritoneal recurrence-free survival. CONCLUSIONS: Molecular diagnosis of peritoneal lavage fluid with the TRC reaction could be a useful prognostic indicator for peritoneal recurrence and survival. Because the TRC reaction is more rapid and simpler than reverse transcriptase-polymerase chain reaction as a format for detecting RNA sequences, it may enhance the genetic diagnosis of cancer micrometastasis and may improve cancer therapy.

A new class of chiral pyrrolidine-pyridine conjugate base catalysts for use in asymmetric Michael addition reactions.

Direct catalytic asymmetric Michael addition reactions of ketones to nitroolefins using newly developed chiral pyrrolidine-pyridine conjugate bases as catalysts are described. The desired 1,4-adducts are obtained in excellent yields with high enantio- and diastereoselectivities.