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6-Methoxy-4-quinolone (6-MOQ, 1 ), an oxidation product derived from 5-methoxyindole-3-acetic acid, is a novel fluorophore, which has several useful characteristics for biomedical analysis. Compound 1 has strong fluorescence with a large Stokes’ shift in aqueous media, and the maximum fluorescence excitation and emission wavelengths are 243 nm and 374 nm, respectively. The molar absorptivity at the maximum excitation wavelength and fluorescence quantum yield in aqueous 10% (v/v) methanol are 32 600 L mol −1 cm −1 and 0.38, respectively. The fluorescence intensity of 1 is scarcely affected by changing the medium pH, showing strong fluorescence from pH 2.0 to 11.0. In addition, 1 is highly stable against light and heat, and no degradation was observed at 60 °C for 3 days with exposure to daylight. As a fluorescent labeling reagent, [(6-methoxy-4-oxo-1,4-dihydroquinolin-3-yl)methyl]amine (6-MOQ-NH 2 , 2 ) was synthesized, and determination of carboxylic acids was demonstrated; 50 pmol of standard propionic acid and isobutyric acid were derivatized, and the obtained S/N ratios for 10 fmol (injection amount) of these two acids were 206 and 164, respectively.

The amount of endogenous melatonin in the individual pineal glands of inbred mice has been determined using reversed-phase micro-high-performance liquid chromatography after precolumn oxidation of melatonin to a compound having strong fluorescence. The fluorescent compound was identified as N-[(6-methoxy-4-oxo-1,4-dihydroquinolin-3-yl)methyl]acetamide. The excitation and emission wavelengths of this compound are 245 and 380 nm, respectively, and the fluorescence intensity is 6.8 times greater than that of melatonin. Molar absorptivity and fluorescence quantum yield of this compound are 46,300[L mol(-1)cm(-1)] and 0.31 (245 nm), respectively. The lower quantification limit of melatonin in biological samples using this precolumn oxidation method is 200 amol, and the calibration curve of spiked melatonin is linear from 200 amol to 50 fmol (r>0.999). The sensitivity of the present method is almost 10 times higher than that of the previous method. The values of endogenous melatonin obtained for ICR, C57BL, BALB/c, and AKR mice are 4.7, 6.1, 7.4, and 18.8 fmol/pineal gland, respectively. The amounts of endogenous pineal melatonin of these strains had not been clearly reported due to the poor enzymatic activities for melatonin biosynthesis; this is the first report that clearly demonstrates the existence of endogenous melatonin in these inbred mice.

We have assessed levels of surface-expressed complement regulatory proteins, decay-accelerating factor (DAF) and membrane cofactor protein (MCP) on cells from patients with hematological malignancies. Neither malignant cells nor unaffected nucleated blood cells from the patients lacked MCP. On the other hand, complete deficiency of DAF was found in 2/10 of non-Hodgkin's lymphoma (NHL), while none of the 38 patients with acute nonlymphocytic leukemia (ANLL) (14 cases), chronic myelogenous leukemia (CML) (6 cases), acute lymphocytic leukemia (ALL) (12 cases) and chronic lymphocytic leukemia (CLL) (6 cases) lacked DAF. The two patients with DAF-negative NHL had no history of paroxysmal nocturnal hemoglobinuria (PNH), and their peripheral blood cells were DAF-positive. One DAF-negative NHL exhibited T cell markers and the other those of B cell. In both cases, treatment of the DAF-negative lymphoma cells with antibody against MCP (M177) followed by Mg(2+)-EGTA-serum resulted in efficient deposition of homologous C3. These results infer that some NHL specifically lack DAF and, through treatment with M177, are targeted by homologous C3.

Having reached the target lesion, macroscopically complete resection was achieved in 28/31 patients (90%). Full thickness resection was confirmed histologically in 26/ 31 cases (84%). Histological complete resection was achieved in 23/31 cases (74%). No perforations were observed during or after resection. In two cases there was a minor bleeding which was controlled with an injection therapy and two patients developed a post-polypectomy syndrome which was managed with antibiotic therapy. Conclusion: Full thickness resection in the lower gastrointestinal tract with the novel FTRD is feasible and effective. Prospective studies are needed to further evaluate the technique and device. Sa1524 Identification of Esophago-Gastric Junction During Per-Oral Endoscopic Myotomy-Benefit of Indocyanine Green Injection Hitomi Minami*, Hajime Isomoto, Haruhiro Inoue, Shigetoshi Urabe, Hiroyuki Shoji, Hiroko Fukuda, Naoyuki Yamaguchi, Kayoko Matsushima, Yuko Akazawa, Ken Ohnita, Fuminao Takeshima, Kazuhiko Nakao Department of Gastroenterology and Hepatology, Nagasaki University Hospital, Nagasaki, Japan; Digestive Disease Center, Showa University Hospital, Tokyo, Japan Introduction: Per-oral endoscopic myotomy (POEM) has become one of the best therapeutic approaches for esophageal achalasia. During POEM, esophagogastric junction (EGJ) can be estimated from inside of the submucosal tunnel by insertion length of the scope, visualizing palisade vessel, and resistance to the endoscope. However, it is often difficult to accurately recognize the EGJ especially in technically challenging cases such as post-operative cases and sigmoid type esophagus. Therefore, we evaluated the usefulness of indocyanine green (ICG) injection at the cardia for precise identification of EGJ during POEM procedure. Material and Method: Eighty-eight cases (male: female 37: 51) aged from 18 to 85 (mean 51.5) who underwent POEM in our institution between August 2010 and November 2014 were enrolled in the study. Mean duration after symptom onset was 11.7 years (1-50, median 5). POEM was performed according to Inoue’s original procedure under general anesthesia and carbon dioxide gas insufflation. For 36 cases since August 2013, 0.5ml of ICG was injected into submucosal layer at the level of cardia prior to POEM as a marking for precise identification of EGJ. Results of the procedure including operation time, subjective symptom score, esophageal manometry study, and complication rate were compared between ICG injected group and noninjection group. Results: There was no significant difference in patients’ background such as dilation grade, duration after onset, X-ray type, and subjective symptom score before the procedure between both groups. Subjective symptom score (Eckardt score; 0.74 vs 0.70, n.s.), endoscopic feature, and mean lower esophageal sphincter pressure (11.8 vs 18.7 mmHg, n.s.) were remarkably improved after the procedure in both groups. No severe complication such as perforation and mediastinitis was seen. Incidence of symptomatic post-POEM GERD, which was easily controlled by normal dose of PPI, did not differ between both groups. Mean operation time of ICG group was no longer than in without ICG injection group (77.5 vs 91.0 min). Discussion & Conclusion: ICG injection enabled us to precisely identify the EGJ and did not affect on the results of the procedure. Especially in the technically challenging cases such as post-operative cases and sigmoid type with severe tortuosity, green color of ICG was effective in discriminating EGJ. Sa1525 Efficacy and Learning Curve of a New Scissor-Like Device for Endoscopic Submucosal Dissection (ESD) in an Established Experimental Setting Helmut Neumann*, Silke Loffler, Markus F. Neurath, Andreas Naegel Department of Medicine 1, University of Erlangen-Nuremberg, Erlangen, Germany Introduction: ESD has become the standard technique for early cancerous lesions, particularly in Asian countries. However, current limitations of ESD include the relative high complication rate, a distinct learning curve and the need of various devices, resulting in substantial costs of the procedure. Aims: To assess the efficacy and learning curve of a new alligator scissor-like device for ESD that will be officially introduced at DDW. Material & Methods: We performed a prospective ex vivo study. Ex vivo porcine models were utilized in the EASIE endoscopic simulator of interventional endoscopy. Artificial lesions, each 2x2 cm in size, were created in fresh ex vivo porcine stomachs at the fundus, corpus and antrum. Two beginners with less than 10 ESD procedures each participated in the study. All study parameters were independently recorded. ESD was performed after marking of the lesions with the closed ESD-instrument, followed by lifting of the mucosa with submucosal injection of saline. Afterwards, circular incision of the lesions was performed with the new ESD-instrument. For resection, the submucosa was grasped with the scissor-like device, elevated and cut. Resection specimens were retrieved to evaluate if all marks were included (R0). Results: Average size of removed lesions was 35 mm. En-bloc resection rate was 100% and R0 resection rate was 86%. After the initial three cases, R0 resection was achieved in all cases. The total procedure time improves significantly during the study (52.4 minutes versus 21.10 minutes; p! 0.05). One perforation occurred which could be managed with endoclips. Endoscopists satisfaction was high throughout all cases. Conclusion: The new scissor-like device for ESD, www.giejournal.org Vol which will be officially introduced at DDW 2015, seems to be a safe and efficient new instrument for ESD which can relatively easy being learned. Further studies should now focus on in vivo studies to confirm these initial results. Sa1526 Natural Orifice Endoscopic Surgery (NOTES) Techniques for Full Thickness R0 Endoscopic Resection of Deep Seated Subepithelial Tumors (SETs): a Single Center Experience Stavros N. Stavropoulos*, Rani J. Modayil, David Friedel, Collin E. Brathwaite, John Allendorf, James H. Grendell Medicine, Winthrop Univ. Hosp., Roslyn, NY Most UGI SETs are GISTs, potentially malignant tumors. Surgery recommended for suspected GISTsR2 cm and endoscopic surveillance of those!2cm. This approach creates a large burden of surgery and endoscopy for small SETs the majority of which are low risk lesions. Several groups have reported ESD to “excavate” muscularis propria (MP) based SETs. However, such enucleation may leave microscopic residual tumor within the MP and is not applicable to tumors with extraluminal component. Recently, pioneering Asian centers reported two endoscopic techniques that achieve R0 en bloc resection of MP-based SETs: Submucosal Tunnel Endoscopic Resection (STER)an offshoot of POEM utilizing the submucosal tunnel method to ensure secure closure of the full-thickness defect in the wall of the GI tract-, and endoscopic full-thickness resection (EFTR)-direct full thickness resection with closure of the defect with clips or sutures. We report the first series of EFTR and STER in the US. Procedures were performed from 4/2012-11/2014. Lesion location/ size, knives, anesthesia, ASA class, length of procedure, closure technique, histologic diagnosis, adverse events (AEs), and length of stay (LOS) were retrieved from a prospectively maintained database. 44 resections (33 EFTRs, 11 STERs) were performed by a gastroenterologist with 5 years POEM and 8 years ESD experience (SNS). Baseline characteristics seen in Table 1. 20 GISTs, 9 leiomyomas, 1 leiomyosarcoma, 6 other tumors, 8 benign “pseudotumors” (not appreciated on pre-op EUS as benign). Mean 23 mm (10-55). Mean resection time 82 minutes (21-400). 84% of patients were admitted mainly for observation. Complete en bloc resection in 41/44 (91%). Piecemeal resection 2 cases (not tumors). 1 morcelized in stomach for extraction (5 cm GIST). AEs: 2 needle decompression of capnoperitoneum, 5 patients 12-48 hr ICU observation, 1 patient with prolonged stay for pleural effusion requiring chest tube drainage. Two patients with challenging resection were done with laparoscopic overview for immediate assistance that was not needed. 1 pt had a 4-5 mm artery adjacent to the tumor identified by EFTR. We elected to complete the resection laparoscopically. EFTR and STER represent a NOTES alternative to laparoscopic wedge resection. We present the first US series with a large number of EFTR cases. Advantages of these techniques include: 1. Scar-less NOTES approach 2. “Wedge” resection of SETs at locations where laparoscopic “wedge” resection is challenging or impossible such as the GE junction, esophagus and gastric cardia 3. Definitive diagnosis and complete resection obviating any further endoscopic surveillance for low risk lesions. The techniques require advanced skills for closure, resection, hemostasis and control of capnoperitoneum and should be pursued only once ESD mastery has been achieved with surgical back-up. Baseline Characteristics ume 81, No. Procedural Data 5S : 2015 GASTROINTESTINAL E Follow-up

To study the localization of myosin in platelets, we utilized polyclonal antibody to heavy chain of platelet myosin. Both immunofluorescence and indirect immunogold staining techniques were employed. (1) In the unactivated platelets, myosin was distributed homogeneously throughout the cytosol. The cytosolic myosin was removed after platelets were treated with Triton X-100. The association of myosin with actin microfilaments in unactivated platelets was minimal. (2) In the surface-activated platelets, myosin was unextractable by Triton X-100. The myosin antibody heavily decorated the actin cable-networks which characterized the activated platelets. (3) In the Triton-unextractable cytoskeleton of both unactivated and activated platelets, we found fine fibrils (about 1-nm wide) that were often associated with immunogolds. These fibrils were similar to purified myosin molecules observed in rotary-shadowed metal replicas and ultrathin sections. These results indicate that cytosolic myosin becomes associated with actin cable-networks after the activation of platelets.