Use of ANGPTL2 mRNA levels in formalin-fixed paraffin-embedded tissues as a biomarker to diagnose gastric cancer and to evaluate the extent of vascular invasion.

With the recent advances in medical technologies, gastric cancer can often be removed with minimally invasive surgical techniques when identified early. Surgery must remove all gastric cancer, since residual cancerous tissue may lead to recurrence. Resected cancerous tissues are pathologically evaluated to determine whether all cancerous areas have been removed, but such assessments are rarely straightforward, and cancer markers could inform such pathological evaluations of cancer. An ideal marker would be identifiable in formalin-fixed paraffin-embedded (FFPE) tumor tissue. The first objective of the present study was to compare levels of angiopoietin-like protein 2 (ANGPTL2) in cancerous and noncancerous areas of FFPE tissues to determine whether ANGPTL2 is a marker relevant to the pathological diagnosis of cancer. The second objective was to evaluate whether ANGPTL2 mRNA is useful as a marker of the extent of vascular invasion of gastric cancer. Out of the 15 patients studied, 12 had a higher ANGPTL2 mRNA levels in cancerous areas compared with noncancerous areas. This finding indicated that ANGPTL2 mRNA is useful as a biomarker for identifying cancerous areas in FFPE tissues, at least for male patients. Spearman's rank correlation analysis showed a significant correlation between the ANGPTL2 mRNA level and the degree of vascular invasion of cancer (r=0.66; P=0.01). In receiver operating characteristic curve analysis of the association between the ANGPTL2 mRNA level and the degree of vascular invasion, the area under the curve was 0.92 (95% confidence interval, 0.78-1.00; P=0.01), indicating a significant association. The present study demonstrates that ANGPTL2 mRNA in FFPE tissues is a potential biomarker that informs the pathological diagnosis of gastric cancer and that ANGPTL2 mRNA may be predictive of vascular invasion, which is an indicator of metastasis in gastric cancer.

Behavioural Response Alteration in Caenorhabditis elegans to Urine After Surgical Removal of Cancer: Nematode-NOSE (N-NOSE) for Postoperative Evaluation.

The technique used for cancer monitoring is essential for effective cancer therapy. Currently, several methods such as diagnostic imaging and biochemical markers have been used for cancer monitoring, but these are invasive and show low sensitivity. A previous study reported that Caenorhabditis elegans sensitively discriminated patients with cancer from healthy subjects, based on the smell of a urine sample. However, whether C. elegans olfaction can detect the removal of cancerous tumours remains unknown. This study was conducted to examine C. elegans olfactory behaviour to urine samples collected from 78 patients before and after surgery. The diagnostic ability of the technique termed Nematode-NOSE (N-NOSE) was evaluated by receiver operating characteristic (ROC) analysis. The ROC curve of N-NOSE was higher than those of classic tumour markers. Furthermore, we examined the change in C. elegans olfactory behaviour following exposure to preoperative and postoperative samples. The results suggest that a reduction in attraction indicates the removal of the cancerous tumour. This study may lead to the development of a noninvasive and highly sensitive tool for evaluating postoperative cancer patients.

Autoclavable physically-crosslinked chitosan cryogel as a wound dressing.

Moist wounds were known to heal more rapidly than dry wounds. Hydrogel wound dressings were suitable for the moist wound healing because of their hyperhydrous structure. Chitosan was a strong candidate as a base material for hydrogel wound dressings because the polymer had excellent biological properties that promoted wound healing. We previously developed physically-crosslinked chitosan cryogels, which were prepared solely by freeze-thawing of a chitosan-gluconic acid conjugate (CG) aqueous solution, for wound treatment. The CG cryogels were disinfected by immersing in 70% ethanol before applying to wounds in our previous study. In the present study, we examined the influence of autoclave sterilization (121°C, 20 min) on the characteristics of CG cryogel because complete sterilization was one of the fundamental requirements for medical devices. We found that optimum value of gluconic acid content of CG, defined as the number of the incorporated gluconic acid units per 100 glucosamine units of chitosan, was 11 for autoclaving. An increased crosslinking level of CG cryogel on autoclaving enhanced resistance of the gels to enzymatic degradation. Furthermore, the autoclaved CG cryogels retained favorable biological properties of the pre-autoclaved CG cryogels in that they showed the same hemostatic activity and efficacy in repairing full-thickness skin wounds as the pre-autoclaved CG cryogels. These results showed the great potential of autoclavable CG cryogels as a practical wound dressing.

A small pancreatic hamartoma with an obstruction of the main pancreatic duct and avid FDG uptake mimicking a malignant pancreatic tumor: a systematic case review.

BACKGROUND: Pancreatic hamartomas are extremely rare and may be misdiagnosed as malignant tumors. We report herein a case of a small, solid-type pancreatic hamartoma. CASE PRESENTATION: A 72-year-old female was incidentally detected pancreatic lesion by ultrasonography. Computed tomography and magnetic resonance imaging revealed a 2.0-cm solid lesion. The main pancreatic duct (MPD) was obstructed by the lesion in the head of the pancreas, and the upstream MPD was dilated. 18F-fluorodeoxyglucose (FDG) accumulated avidly in the lesion and increased in FDG intensity from the early to the delayed images. The histopathological studies confirmed the diagnosis of pancreatic hamartoma. Immunohistochemically, the cell membrane of the accessory glands and ducts showed homogeneous expression of glucose transporter type I and hexokinase II. CONCLUSION: Pancreatic hamartomas causing dilatation of the MPD are extremely rare, and this appears to be the first case of a hamartoma to take up FDG avidly. It was a rare occurrence and should be noted that pancreatic hamartomas can cause an obstruction of the MPD and show avid FDG uptake, thereby mimicking malignant pancreatic tumors.

Necrotic regions are absent in fiber-shaped cell aggregates, approximately 100 µm in diameter.

Microscopic, fiber-shaped cell aggregates, have been used as building blocks for fabricating macroscopic three-dimensional tissue architectures, in the field of tissue engineering. In this study, we examined the occurrence of necrotic regions in the most widely used, fiber-shaped cell aggregates, approximately 100 µm in diameter. Alginate hydrogel hollow microfibers were used as templates for the cell aggregates. We demonstrated negligible necrotic region formation occurred in the cell aggregates formed in the hollow microfibers. Furthermore, we improved on previously-reported methods for preparing the hollow microfibers to avoid common microfiber tangling during the fiber preparation process.

Correlations of (18)F-fluorothymidine uptake with pathological tumour size, Ki-67 and thymidine kinase 1 expressions in primary and metastatic lymph node colorectal cancer foci.

OBJECTIVE: To examine correlations of (18)F-fluorothymidine (FLT) uptake with pathological tumour size and immunohistochemical Ki-67, and thymidine kinase 1 (TK-1) expressions in primary and metastatic node colorectal cancer foci. METHODS: Thirty primary cancers (PCs) and 37 metastatic nodes (MNs) were included. FLT uptake was assessed by visual scores (non-visible: 0-1 and visible: 2-4), standardized uptake value (SUV), and correlated with size, Ki-67, and TK-1. SUV was measured in visible lesions. FLT heterogeneity was assessed by visual scores (no heterogeneous uptake: 0 and heterogeneous uptake: 1-4). RESULTS: Forty-two lesions were visible. The visible group showed significantly higher values than the non-visible group in size, Ki-67, and TK-1 (each p < 0.05). Size correlated significantly with visual score (PC; ρ = 0.74 and MN; ρ = 0.63), SUVmax (PC; ρ = 0.49, and MN; ρ = 0.76), and SUVmean (PC; ρ = 0.40 and MN; ρ = 0.76) (each p < 0.05). Visual score correlated significantly with size (ρ = 0.86), Ki-67max (ρ = 0.35), Ki-67mean (ρ = 0.38), TK-1max (ρ = 0.35) and TK-1mean (ρ = 0.25) (each p < 0.05). No significant correlations were found between FLT uptake and Ki-67 or TK-1 in 42 visible lesions (each p > 0.05). Heterogeneous FLT uptake was noted in 73 % (22/30) of PCs. CONCLUSION: FLT uptake correlated with size. Heterogeneous FLT distribution in colorectal cancers may be one of the causes of weak or lack of FLT uptake/Ki-67 or TK-1 correlation. KEY POINTS: FLT uptake correlated well with tumour size in colorectal cancer. Weak or lack of FLT uptake/Ki-67 and TK-1 correlations were observed. Immunohistochemical Ki-67 and TK-1 expressions are not always correlated with FLT uptake.

Diagnosis of pancreatic neoplasms using a novel method of DNA methylation analysis of mucin expression in pancreatic juice.

Mucins (MUC) play crucial roles in carcinogenesis and tumor invasion in pancreatic ductal adenocarcinoma (PDAC) and intraductal papillary mucinous neoplasms (IPMNs). Our immunohistochemistry (IHC) studies have shown a consensus position on mucin expression profiles in pancreatic neoplasms as follows: MUC1-positive but MUC2-negative expression in PDACs; MUC1-negative but MUC2-positive expression in intestinal-type IPMNs (dangerous type); MUC1-negative and MUC2-negative expression in gastric-type IPMNs (safe type); High MUC4 expression in PDAC patients with a poor outcome; and MUC4-positive expression in intestinal-type IPMNs. We also showed that three mucin genes (MUC1, MUC2 and MUC4) expression in cancer cell line was regulated by DNA methylation. We have developed a novel 'methylation-specific electrophoresis (MSE)' method to analyze the DNA methylation status of mucin genes by high sensitivity and resolution. By using the MSE method, we evaluated pancreatic juice samples from 45 patients with various pancreatic lesions. The results were compared with final diagnosis of the pancreatic lesions including IHC of mucin expression in the paired pancreatic tissues. The results indicated that the DNA methylation status of MUC1, MUC2 and MUC4 in pancreatic juice matched with the mucin expression in tissue. Analyses of the DNA methylation status of MUC1, MUC2 and MUC4 were useful for differential diagnosis of human pancreatic neoplasms, with specificity and sensitivity of 87% and 80% for PDAC; 100% and 88% for intestinal-type IPMN; and 88% and 77% for gastric-type IPMN, respectively. In conclusion, MSE analysis of human pancreatic juice may provide useful information for selection of treatment for pancreatic neoplasms.

Effect of chitosan-gluconic acid conjugate/poly(vinyl alcohol) cryogels as wound dressing on partial-thickness wounds in diabetic rats.

We previously developed chitosan cryogels from chitosan-gluconic acid conjugate without using toxic additives for wound care. In this study, we improved physiological characteristics of the previous cryogels by incorporating poly(vinyl alcohol) that also form cryogels. Mechanical strength of the cryogels was more than two times higher than that of the previous cryogels. Furthermore, the incorporation of poly(vinyl alcohol) enhanced water retention and resistance to degradation of the gels by lysozyme. The cryogels retained the favorable biological properties of the previous cryogels that they accelerate infiltration of inflammatory cells into wound sites. Time period for repairing 50 % of initial area of partial-thickness skin wound treated with the cryogels (4.0 ± 1.1 days) was shorter than those with gauze (6.5 ± 0.3 days) or a commercial hydrogel dressing (5.7 ± 0.3 days). Finally, we confirmed that incorporation of basic fibroblast growth factor into the cryogels was effective to further accelerate wound healing (2.7 ± 1.0 days). These results demonstrate that the cryogels in this study are promising for wound care.

Diagnostic performance of ¹8F-fluorothymidine PET/CT for primary colorectal cancer and its lymph node metastasis: comparison with ¹8F-fluorodeoxyglucose PET/CT.

PURPOSE: To examine the diagnostic performance of (18)F-fluorothymidine (FLT) PET/CT in primary and metastatic lymph node colorectal cancer foci in comparison with (18)F-fluorodeoxyglucose (FDG) PET/CT. METHODS: The study population comprised 28 patients with 30 newly diagnosed colorectal cancers who underwent surgical resection of the primary lesion and regional lymph nodes after both FLT and FDG PET/CT. The associations between SUVmax levels and pathological factors were evaluated using the Mann-Whitney U or Kruskal-Wallis test. Differences in diagnostic indexes for detecting nodal metastasis between the two tracers were estimated using the McNemar exact or χ(2) test. RESULTS: All 30 primary cancers (43.0 ± 20.0 mm, range 14 - 85 mm) were visualized by both tracers, but none of the FLT SUVmax values exceeded the FDG SUVmax values in any of the primary cancers (6.6 ± 2.4 vs. 13.6 ± 5.8, p < 0.001). The sensitivity, specificity and accuracy for detecting nodal metastasis were 41% (15/37), 98.8% (493/499) and 94.8% (508/536) for FDG PET/CT, and 32% (12/37), 98.8% (493/499) and 94.2% (505/536) for FLT PET/CT, respectively. The sensitivity (p = 0.45), specificity (p = 0.68) and accuracy (p = 0.58) were not different between the tracers. Nodal uptake of FLT and FDG was discordant in 7 (19%) of 37 metastatic nodes. There were ten concordant true-positive nodes of which six showed higher FDG SUVmax and four showed higher FLT SUVmax, but the difference between FDG and FLT SUVmax was not significant (5.56 ± 3.55 and 3.62 ± 1.45, respectively; p = 0.22). CONCLUSION: FLT has the same potential as FDG in PET/CT for the diagnosis of primary and nodal foci of colorectal cancer despite significantly lower FLT uptake in primary foci.

High FDG and low FLT uptake in a thyroid papillary carcinoma incidentally discovered by FDG PET/CT.

We report a 58-year-old man whose incidentally discovered papillary thyroid carcinoma in the left lobe showed high FDG and low FLT uptake on PET/CT. The SUVmax was 19.7 for FDG and 3.0 for FLT. The Ki-67 labeling index of the tumor was 1.9%. Thus, the low FLT uptake might be attributed to the low proliferative activity of this cancer.

[Two cases of circumferential rectal invasion from prostatic cancer].

We report two cases of prostatic cancer invading the rectum with circumferential rectal wall thickening. In both cases, reddish and uneven edematous mucosa was observed by colonoscopy and the boundary of the lesion was indistinct. No mass or ulcer formation was not observed. CT scan showed irregular prostatic swelling and circumferential rectal wall thickening adjacent to the prostate. The rectal biopsy specimen showed moderately to poorly differentiated adenocarcinoma, and it was positive on PSA immunostaining. Although rectal invasion of prostatic cancer is relatively rare, it is important to be aware that prostatic cancer can cause circumferential stenosis of the rectum.

[A retrospective endoscopic case of adenoid cystic carcinoma of the esophagus].

We report a case of a 64-year-old Japanese man with adenoid cystic carcinoma. An elevated lesion covered by intact epithelium in the thoracic esophagus was found in September, 2007 and been followed. After dysphagia appeared follow-up endoscopy was performed in January, 2010, and morphological change into a protruding tumor was recognized. Since adenoid cystic carcinoma was detected by endoscopic biopsy, the patient underwent esophageal resection. The resected specimen showed a cribriform pattern and a certain amount of mucous substance which was positive for Alcian blue, within a solid nest. The tumor cells were positive for S-100 protein and negative for αSMA, so the tumor was diagnosed as an adenoid cystic carcinoma.

[A case of eosinophilic gastroenteritis with ring-like discoloration in the lower end of the esophagus].

We report the case of a 15-year-old Japanese boy with eosinophilic gastroenteritis. The patient complained of abdominal pain and watery diarrhea and had a history of allergic rhinitis. Laboratory data on admission showed leukocytosis with remarkable eosinophilia. Microscopic examination of the biopsied specimens taken from the esophagus, stomach, duodenum, lower ileum and colon showed eosinophilic infiltration. Especially in the lower esophagus, there was a ring-like discoloration with remarkable eosinophil infiltration. We diagnosed eosinophilic gastroenteritis and his clinical symptoms and eosinophilia improved following starting corticosteroid therapy. After 5 months therapy with prednisolone, discoloration of upper digestive tract disappeared. There have been no reports describing discoloration in the lower end of the esophagus seen by gastroscopy.

Clinical significance of primary lesion FDG uptake for choice between oesophagectomy and endoscopic submucosal dissection for resectable oesophageal squamous cell carcinomas.

OBJECTIVES: To correlate primary oesophageal squamous cell carcinoma (SCC) (18)F-fluoro-deoxyglucose (FDG) uptake with pathological factors and examine its significance regarding choice of therapy. METHODS: We retrospectively examined the factors affecting visible and non-visible FDG uptake in 37 primary lesions in 32 oesophageal SCC patients who underwent PET/CT before oesophagectomy or endoscopic submucosal dissection (ESD). We divided the lesions into pathological depth invasion ≥sm2 oesophagectomy (n = 18) and ≤sm1 ESD (n = 19) indicated groups and compared the diagnostic accuracy of FDG-PET with that of endoscopic ultrasound (EUS) performed for 23 superficial lesions to discriminate between these groups. RESULTS: There were 17 visible and 20 non-visible lesions. The lesion visibility was significantly higher in the larger (≥40 mm), non-flat type, more deeply invaded, positive vascular invasion (P < 0.001 each), positive nodal metastasis (P = 0.04) and higher Glut-1 score (P = 0.005) tumour groups. When the visible and non-visible lesions indicated a need for oesophagectomy and ESD respectively, the sensitivity, specificity and accuracy of oesophagectomy were 94% (17/18), 100% (19/19) and 97% (36/37) and those of EUS were 75% (3/4), 79% (15/19) and 78% (18/23) respectively. CONCLUSIONS: Primary lesion FDG visibility can be one of the indicators for choosing between oesophagectomy and ESD for resectable oesophageal SCCs.

MUC1 mucin expression in follicular dendritic cells and lymphoepithelial lesions of gastric mucosa-associated lymphoid tissue lymphoma.

Membrane-associated mucin MUC1 is expressed in various adenocarcinoma cells and active T lymphocytes. We tried to find out whether MUC1 is expressed in gastric mucosa-associated lymphoid tissue (MALT) lymphoma lesion. MUC1 was not expressed in infiltrating T lymphocytes; however, MUC1 was found on the cell surface of follicular dendritic cells (FDC) of germinal centers and in the epithelial cytoplasm of lymphoepithelial lesion (LEL) of the lymphoma, which were immunohistochemically detected by monoclonal antibodies DF3 and MY.1E12. MUC1 was also expressed in the FDC of control cases (gastrectomy specimen containing reactive lymphoid follicles, n = 10, MUC1/ DF3, 100%; MUC1/MY.1E12, 40%), and FDC in MALT lymphomas (n = 59) showed lower MUC1 expression rates (MUC1/ DF3, 32%; MUC1/MY.1E12, 0%) than the control (P < 0.001). Lymphoepithelial lesion in the low-grade MALT lymphomas (n = 23) showed a higher MUC1/DF3 expression rate (30%) than those in the high-grade MALT lymphomas (n = 36; 6%; P < 0.05). T lymphocytes in the surface mucosa were more frequent in MALT lymphoma (91.4 +/- 80.6/unit area) than those in the control (20.0 +/- 23.6) (P < 0.001). S100-positive dendritic cells around LEL were more frequent in the low-grade (19.0 +/- 9.4/unit area) than in the high-grade (11.7 +/- 9.7) (P < 0.005). This study demonstrated MUC1 mucin expression on FDC for the first time. Mucosa-associated lymphoid tissue lymphoma, especially low-grade, shows immunologically active state, where FDC MUC1 expression may be suppressed by some factors released from lymphoma cells. Further study to elucidate the pathogenetic role of MUC1 in MALT lymphoma is necessary.

Expression of MUC1 and MUC2 mucins in extrahepatic bile duct carcinomas: its relationship with tumor progression and prognosis.

Our previous immunohistochemical studies in the pancreas, intrahepatic bile duct, and ampulla of Vater demonstrated that an invasive carcinoma with a poor outcome showed a pattern of MUC1 (membrane-bound mucin) positive and MUC2 (intestinal-type secretory mucin) negative, whereas many of the non-invasive tumors with favorable outcome showed a pattern of MUC1 negative and MUC2 positive. The aim of this study is to compare the expression profiles of MUC1 and MUC2 mucins in extrahepatic bile duct carcinomas to gain insight into the relationship between the biological nature of the carcinomas and the role of mucins. We examined the expression profiles of MUC1 of different glycoforms and MUC2 in 60 extrahepatic bile duct carcinomas using immunohistochemistry.The expression of MUC1/CORE (core peptide of MUC1), MUC1/DF3 (core peptide of MUC1 with sialyl oligosaccharides) and MUC1/MY.1 E12 (sialylated MUC1) showed a significant relationship with tumor progression factors such as poor differentiation, deep invasion, lymph node metastasis, lymphatic invasion or perineural invasion. In contrast, the expression of MUC1/HMFG-1 (fully glycosylated MUC1) did not show a significant relationship with the tumor progression factors. In the different glycoforms of MUC1 examined, the expression of MUC1/DF3 and MUC1/MY.1E12 was related with the poor outcome of the patients. In contrast, the expression of MUC2 was inversely related with the tumor progression factors and poor outcome. In the 52 patients with advanced tumors, only MUC1/DF3 high expression correlated with poor prognosis. In conclusion, MUC1/DF3 was the most useful prognosis indicator among the various glycoforms of MUC1 mucins.