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Background and Aim: Endoscopic sphincterotomy (ES) combined with large balloon dilation (ESLBD) can be useful for extracting large and multiple bile duct stones. Although there are many studies on the feasibility and short-term outcome, there are few reports about mid- to long-term outcome after ESLBD. The aim of our study is to prospectively evaluate the mid-term outcome of ESLBD., Methods: One hundred eighty-three patients who underwent ESLBD between November 2006 and May 2012 were included. The patients were followed up periodically after the procedure until April 2013. Papillary dilation was performed at the time of initial ES or prior ES. Early and late adverse events and stone recurrence were evaluated in this study., Results: The patients' mean age was 76.6 ± 10.7 years. Surgically altered anatomy was present: Billroth I gastrectomy (2), Billroth II gastrectomy (13), and gastrectomy with Roux-en Y reconstruction (18). Seventy-eight (42.6%) patients had periampullary diverticulum. Prior ES had been performed in 40 (21.9%) patients. The mean follow-up period was 43.5 ± 19.7 months (range 11-78). Eight (4.4%) patients had stone recurrence. There was rare stone recurrence after initial ESLBD treatment and native gastrointestinal anatomy. Univariate and multivariate analyses showed that prior ES and previous history of stone recurrence were predictive of stone recurrence (P < 0.001)., Conclusion: At mid-term outcome, ESLBD is associated with a low rate of recurrent bile duct stones, although long-term follow up is needed., (© 2014 Journal of Gastroenterology and Hepatology Foundation and Wiley Publishing Asia Pty Ltd.)

Aim: To investigate how complete laparoscopic anterior resection with natural orifice specimen extraction (NOSE), as a novel minimally invasive surgery, compares to conventional laparoscopic surgery., Methods: Twenty patients who underwent complete laparoscopic anterior resection with NOSE and 50 patients who underwent laparoscopic assisted anterior resection by the conventional method between 2011 and 2012 were studied. Selection for complete laparoscopic anterior resection with NOSE was decided on the basis of tumor size, localization of the tumor, and body mass index. Outcomes related to surgery, including operation time, postoperative wound pain, hospital stay after surgery, the number of totally dissected lymph nodes, postoperative complications (suture failure and wound infection), and anal function, were reviewed retrospectively. Anal function was assessed at 3 and 6 mo after surgery using the Wexner fecal incontinence scoring system., Results: Complete laparoscopic resection with NOSE was performed to completion in all 20 patients. There was no patient emergency that required conversion to conventional laparoscopic surgery or open surgery. The comparison between complete laparoscopic resection with NOSE and conventional laparoscopic surgery showed no significant differences in the maximal diameter of the tumor, number of totally dissected lymph nodes, bleeding volume, mean operation time, time to start of oral ingestion, postoperative hospital stay, and postoperative complications. On the other hand, with regard to pain after epidural anesthesia, the total usage of analgesia in this novel surgical technique was 1.85 ± 1.8 times, whereas it was 5.89 ± 2.86 in conventional laparoscopic surgery (P < 0.001). The postoperative pain period was 1.9 ± 1.9 d in this novel surgical technique, whereas it was 3.43 ± 1.41 d in conventional laparoscopic surgery (P < 0.004). In complete laparoscopic surgery with NOSE, the mean postoperative follow-up period was 20 mo (range: 12-30 mo). Neither local recurrence nor remote metastasis was observed during the follow-up period., Conclusion: Complete laparoscopic anterior resection using NOSE does not require any incision and has excellent cosmetic properties, with mitigated postoperative pain.

Background/aims: To achieve R0 resection, pancreaticoduodenectomy with right-side half dissection of the superior mesenteric artery nerve plexus is performed for pancreatic cancer with extrapancreatic nerve plexus invasion in many facilities. However, this cancer mainly spreads behind the superior mesenteric artery., Methodology: Forty-two patients underwent pancreaticoduodenectomy with right-oblique posterior dissection of the superior mesenteric artery nerve plexus from the 4 to 10 o'clock position for pancreatic ductal adenocarcinoma. The cancer spread was evaluated using preoperative multi-detector computed tomography and postoperative pathological examination., Results: Thirty-one patients (73.8%) showed extrapancreatic nerve plexus invasion on multi-detector computed tomography. In 20 patients (47.6%), the tumor extended within 5 mm of the superior mesenteric artery, ranging between the 4-10 o'clock position in 19 (95.0%) patients. Although pathological examination revealed that the cancer infiltrated within 3 mm of the superior mesenteric artery margin in 17 (54.8%) patients with extrapancreatic nerve plexus invasion, R0 resection was achieved in 95.2% of cases. Six patients (14.3%) experienced postoperative diarrhea requiring administration of antidiarrheal agents., Conclusions: Pancreatic head cancer spreads mainly right-posterior of the superior mesenteric artery; and therefore, right-oblique posterior dissection is a logical procedure to achieve negative margin resection with complete clearance of nerve plexus involvement.

Only hepatic metastasectomy has been shown to have a therapeutic effect for hepatic metastases of colorectal cancer if the primary tumor can be removed radically according to the 2014 guidelines of the Japanese Society for Cancer of the Colon and Rectum. However, only a few patients with hepatic metastases are candidates for metastasectomy due to tumor factors other than the hepatic metastases. Furthermore, hepatic metastasectomy is frequently judged to be impossible in older patients due to underlying diseases and surgical tolerability. In such cases, therefore, treatment is often difficult. Herein, we report 2 patients with hepatic metastases of colorectal cancer treated with radiofrequency ablation, raising the possibility of local control in the liver.

According to the 2014 guidelines for the treatment of Stage IV colorectal cancer, surgical therapy is recommended if R0 resection is possible by resecting both distant metastases and the primary tumor. However, there was no clear evidence regarding the treatment of non-regional lymph node metastases and recurrent lymph node metastases. Thus, we evaluated cases of non-regional lymph node metastases and recurrent lymph node metastases at our hospital from 2005 to the present date. The 5-year overall survival (OS) rate was 50.0%. There was no significant difference in the survival rate between synchronous and metachronous metastases. The survival rate increased significantly in cases that received additional local treatment compared to that in cases that did not receive additional treatment (p=0.013). Multivariate analysis based on the 5-year OS rate of 50.0% revealed a statistically significant difference only in disease free interval (DFI). Although no predictive factor for OS was identified, DFI was considered to be a critical factor. Performing chemotherapy during the perioperative period and determining the appropriate timing of resection based on the response to the chemotherapy and presence/absence of metastases in other organs are expected to increase the survival rate.

An 18-week-old female breeding Yorkshire pig displayed symptoms of astasia and subsequently died. Histologically, severe disseminated suppurative meningoencephalomyelitis was detected, as were numerous myocardial microabscesses. Gram-positive cocci were detected in these suppurative lesions, and these cocci reacted with an antibody against Streptococcus C group species. Gram-positive cocci were isolated from the liver, spleen, kidney, heart, lungs, pleural abscess and articular fluid of the right tarsal joint. The isolates were β-hemolytic, categorized into Lancefield group C and were identified as Streptococcus dysgalactiae subspecies equisimilis by analysis of the 16S ribosomal DNA sequence. This is the first report of systemic S. equisimilis infection in a pig with severe disseminated suppurative meningoencephalomyelitis.

Multiplanar reconstruction (MPR) images are used for assessing horizontal and vertical extent of hilar cholangiocarcinoma, while peroral cholangioscopy (POCS) is used for diagnosing ductal spread and mapping biopsy. We fused conventional 3-dimensional computed tomography (3DCT) with MPR images and POCS findings for preoperative assessment of cholangiocarcinoma. The extent of cancer was assessed using MPR images, which were plotted and fused onto 3DCT cholangiography. In addition, the results of mapping biopsy performed under POCS were marked on virtual endoscopic imaging and transferred onto a 3DCT image. Once an angiographic CT image was fused, a multi-3DCT image was created. The incision line was determined based on these images. Multi-3DCT images were created for 13 patients with hilar cholangiocarcinoma. Of 10 patients who underwent POCS, superficial spread was observed in two. Resection was performed in 12 patients. In two cases, the cut end of the intrahepatic bile duct was positive, resulting in 83.3% diagnostic accuracy for horizontal spread. In all patients, the estimated number of bile ducts was the same as the number of the actual resections. R0 resection was achieved in 10 patients (83.3%). Multi-3DCT imaging proved useful in diagnosing longitudinal ductal spread of hilar cholangiocarcinoma., (© 2014 Japanese Society of Hepato-Biliary-Pancreatic Surgery.)

To date, several endoscopic ultrasonography (EUS)-guided interventions have been reported. Of these interventions, EUS-guided pancreatic duct (PD) intervention seems to be the most difficult and challenging procedure to accomplish. We provide suggestions for EUS-guided PD intervention, which includes PD stenting and the rendezvous technique following EUS-guided pancreatography. PD stenting is performed between the PD and the gastrointestinal tract (stomach, duodenum, or jejunum). The rendezvous technique is performed using a guidewire across the papilla or anastomotic site for retrograde stent insertion. EUS-guided PD intervention is still challenging because it is not established yet. Thus, although EUS-guided PD intervention seems to be promising, it should be performed in selected patients such as those of failed standard endoscopic retrograde cholangiopancreatography., (© 2013 Japanese Society of Hepato-Biliary-Pancreatic Surgery.)

The mucosal immune system provides the first line of defense against inhaled and ingested pathogenic microbacteria and viruses. This defense system, to a large extent, is mediated by the actions of secretory IgA. In this study, we screened 140 strains of lactic acid bacteria for induction of IgA production by murine Peyer's patch cells. We selected one strain and named it Lactobacillus plantarum AYA. We found that L. plantarum AYA-induced production of IL-6 in Peyer's patch dendritic cells, with this production promoting IgA(+) B cells to differentiate into IgA-secreting plasma cells. We also observed that oral administration of L. plantarum AYA in mice caused an increase in IgA production in the small intestine and lung. This production of IgA correlated strongly with protective ability, with the treated mice surviving longer than the control mice after lethal influenza virus infection. Our data therefore reveals a novel immunoregulatory role of the L. plantarum AYA strain which enhances mucosal IgA production and provides protection against respiratory influenza virus infection.

Objectives: Shear wave elastography is a novel noninvasive method for assessing liver fibrosis by measuring liver stiffness. This study was conducted to evaluate how pathologic changes could have an impact on measured elasticity values in both resected hepatocellular carcinomas and adjacent liver tissue., Methods: Intraoperative shear wave elastography was performed in 7 patients who underwent liver resection at our institution; 7 hepatocellular carcinomas and adjacent liver tissue were subjected to elastographic measurements. A total of 48 circular regions of interest (ROIs; 3-8 mm in diameter) were located in the hepatocellular carcinomas (n = 37) and adjacent liver tissue (n = 11), and mean stiffness values were obtained from each ROI. All of the histologic images corresponding to the 48 ROIs after surgery were transformed into digital microscopic images by a scanning system, and histologic parameters, such as the proportions of nuclear areas, fatty areas, fibrous areas, and vessel areas, were quantitatively assessed. Relationships between the mean stiffness and the histologic parameters were investigated by the mixed effects model., Results: By univariate analysis, the proportions of collagen fiber areas (P = .039), fibrous areas (P = .045), hepatocellular nuclear areas (P = .045), and nuclear areas other than hepatocellular and lymphoplasmacytic areas (P = .039) showed statistically positive associations with mean stiffness values. Multivariate analysis indicated that the proportion of collagen fiber areas was the strongest pathologic determinant of mean stiffness (P = .008), with hepatocellular nuclear areas also having a significant effect (P = .010)., Conclusions: Fibrosis predictably affects elastographic estimation, but hepatocellular density (ie, hepatocellular nuclear areas) also alters elastographic assessment.

Objective: To determine if the POSSUM, SOFA, MPI, and SAS scores provide a better measure of severity for patients with prognostic factors undergoing surgery for colorectal perforation., Subjects: Fifty-nine patients who underwent surgery between 1996 and 2012., Methods: We retrospectively reviewed background factors, blood and physiological test results, and intraoperative findings of patients who survived and those who died. We also compared the POSSUM, SOFA, MPI, and SAS scores. Multivariate analysis was performed for factors that were significant by univariate analysis, and selected factors were used to produce a predictive prognostic model., Results: Univariate analysis revealed significant differences in age, anticoagulant/steroid administration, serum creatinine level, PF ratio, base excess (BE), chest radiography, pulse rate, and severity of peritoneal soiling. Age, serum creatinine level, pulse rate, and severity of peritoneal soiling were selected for multivariate analysis; only pulse rate was significantly different. There were significant differences between the two groups in POSSUM PS, OSS, SOFA, and MPI scores, and a comparison in terms of the ROC curve showed that our model had the highest peak; the area under the curve was 94.8% compared with 70-80% for the other systems, suggesting that our model is better than those systems., Conclusions: POSSUM and SOFA are valid methods of evaluating risk from colorectal perforation, but our study revealed addition risk factors: (1) the PF ratio and BE, which are not included in POSSUM; (2) the pulse rate and severity of peritonitis, which are not included in SOFA; and (3) anticoagulant/steroid hormone administration., (Copyright © 2014 Surgical Associates Ltd. Published by Elsevier Ltd. All rights reserved.)

Endoscopic ultrasonography-guided (EUS)-guided pancreatic interventions have gained increasing attention. Here we review EUS-guided pancreatic duct (PD) access techniques and outcomes. EUS-guided PD intervention is divided into two types, antegrade and rendezvous techniques, following EUS-guided pancreatography. In the antegrade technique, pancreaticoenterostomy is carried out by stent placement between the PD and the stomach, duodenum, or jejunum. Transenteric antegrade PD stenting is conducted by stent placement, advancing anteriorly into the PD through the pancreatic tract. The rendezvous technique is carried out by using a guidewire through the papilla or anastomotic site for retrograde stent insertion. In terms of EUS-guided PD stenting, 11 case reports totaling 75 patients (35 normal anatomy, 40 altered anatomy) have been published. The technical success rate was greater than 70%. Early adverse events, including severe hematoma and severe pancreatitis,occurred in seven (63.6%) of 11 reports. Regarding the rendezvous technique, 12 case reports totaling 52 patients (22 normal anatomy, 30 altered anatomy) have been published. The technical success rate ranged from 25% to 100%. It was 48% in one report that involved more than 20 cases. Once stents were placed, all patients became free of symptoms. Early mild adverse events occurred in four (36.4%) of 11 reports. In conclusion, although it can be risky because of possible serious or even fatal adverse events, including pancreatic juice leakage, perforation and severe acute pancreatitis, EUS-PD access seems to be promising for treating symptomatic pancreatic diseases caused by PD stricture and pancreaticoenterostomy stricture., (© 2013 The Authors. Digestive Endoscopy © 2013 Japan Gastroenterological Endoscopy Society.)

Objectives: Clinically relevant postoperative pancreatic fistula (POPF) after pancreatoduodenectomy is often accompanied by bacterial infection. To elucidate the mechanism of bacterial infection associated with POPF, we investigated the relationship between POPF and bacteria isolated from ascitic fluid and removed drains., Methods: Subjects were 101 patients who had undergone pancreatoduodenectomy. Microbial culture was performed using ascitic fluid obtained from drains. We also compared the isolated bacteria from removed drains on postoperative day (POD) 4 and after POD 7., Results: In 23 patients (22.8%), microbial cultures were already positive on POD 1, although purulent discharge was not observed. Among patients with grade B/C POPF, bacteria were detected on POD 1 in 53.8%; these isolated bacteria were the same as those isolated after POPF formation. In contrast, only 7.7% of patients with grade A POPF were positive on POD 1. The number of bacteria isolated from drains removed after POD 7 significantly increased compared with those isolated from drains removed on POD 4., Conclusions: Bacterial contamination in ascitic fluid may be an initiating event that leads to the development of clinically relevant POPF. Therefore, it is important to perform both the administration of the appropriate antibiotics and early drain removal.

Purpose: Detailed procedures for optimal therapeutic hypothermia (TH) have yet to be established. We examined how duration of well-controlled core temperature within the first 24 hours after cardiac arrests (CA) correlated with neurological outcomes of successfully resuscitated out-of-hospital CA (OHCA) patients., Methods: OHCA patients who survived over 24 hours and treated with TH were included. Core temperature was measured every hour. Physicians intended to maintain temperature at 33 °C ± 1 °C for 24 hours. Cerebral performance categories (CPC) of patients at 6 months were recorded and patients were retrospectively divided into favorable (CPC1,2) and poor (CPC3-5) neurological outcome groups. Total time while the core temperature reached to 33 °C ± 1 °C within the first 24 hours after CA was measured and this duration was defined that of well-controlled temperature. receiver-operating characteristic analysis was performed on duration of well-controlled temperature to select the optimal cutoff value. Neurological outcome predictors were investigated by logistic regression analysis., Results: Fifty-six patients were included. Optimal cutoff value of duration of well-controlled temperature was 18 hours. Ratio of male sex, witnessed by emergency medical service (EMS) personnel, first electrocardiogram as shockable, and duration of well-controlled core temperature ≥ 18 h of favorable neurological outcome group (n = 21) were significantly larger than that of poor neurological outcome group (n = 35). Logistic regression analysis identified "witnessed by EMS", "performed bystander CPR," and "the duration ≥ 18 h" as independent predictors of favorable neurological outcome., Conclusion: TH maintained at target temperature of 33 °C ± 1 °C over 18 hours independently correlated with favorable neurological outcome. Therefore, stable core temperature control may improve neurological outcome of successfully resuscitated OHCA., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: The effects of ablation with various settings of powers and times using a newly developed radiofrequency (RF) ablation device, the Habib™ EndoHPB catheter, are not well known. In the present study, we examined the effects of a novel RF ablation catheter using resected fresh pig livers and evaluated the macro- and microscopic effects of RF ablation under various conditions., Materials and Methods: The RF application was performed step by step at 5, 10, 15, and 20 W power and 60, 90, 120 s, respectively. Macroscopic and microscopic findings of the ablation area were evaluated at each setting., Results: The mean lengths of the short axis of the ablation area at 10 W and 60, 90 and 120 s were 8.0 ± 1.0, 8.3 ± 1.2, and 9.7 ± 0.6 mm, respectively. The mean lengths of the long axis at 10 W power and 60, 90 and 120 s were 20.3 ± 0.6, 21.3 ± 1.6, and 28.3 ± 2.1 mm, respectively. Although the lengths of the short and long axes at 5 and 10 W increased gradually with power, there were no obvious differences in either short or long axis lengths between 15 and 20 W. Of all the settings, only at 5 W and 60 and 90 s did the long axis of the ablation show separate areas around the 2 ring electrodes., Conclusions: Although other sequelae including hemorrhage, pancreatitis, acute inflammatory changes, perforation and late fibrosis could not be investigated in our ex-vivo pig model, our study clarified the relationship between ablation powers and times and the effects concerning depth and longitudinal spread of ablation. Although the clinical ablation setting at 7-10 W power and 2 min is suitable, ultimately the ablation power and time should be adjusted according to the size of masses using examples from the present results.

Type IV-A choledochal cysts (CCs) are a congenital biliary anomaly which involve dilatation of the extrahepatic and intrahepatic bile ducts. We present the case of a 30-year-old woman with type IV-A CC, on whom three-dimensional computed tomography (3D CT) and virtual endoscopy were performed. 3D CT revealed partial dilatation in the posterior branch of the intrahepatic bile duct and a relative stricture between it and the extrahepatic bile duct. Virtual endoscopy showed that this stricture was membrane-like and separated from the surrounding blood vessels. Based on these image findings, complete cyst resection, bile duct plasty for the stricture, and hepaticojejunostomy were safely performed. To the best of our knowledge, there are no reports of imaging by virtual endoscopy of the biliary tract which show the surrounding blood vessels running along the bile duct.

Background/aims: Single nucleotide polymorphism (SNP) of the genes for ATP-binding cassette transporters is related to the side effects of anticancer drugs and that of drug metabolism-related enzyme genes is involved in the activation of gemcitabine (GEM)., Methodology: Forty eight patients treated with adjuvant GEM chemotherapy after pancreatic cancer resection was examined for the SNP of multidrug-resistance 1 (MDR1) 2677, MDR1 3435, breast cancer resistance protein (BCRP) 421, ribonucleotide reductase M1 (RRM1)(-)524, RRM1(-)37 and deoxycytidine deaminase (CDA) 208. We divided the patients according to normal group: patients homozygous for a wild-type allele or heterozygous for a mutant allele and mutant group: those homozygous for a mutant allele. Both groups were compared regarding the outcome and the occurrence and severity of side effects., Results: MDR1 2677, MDR1 3435, BCRP421, RRM1(-) 524, RRM1(-) 37 and CDA mutant groups comprised 37.5, 31.3, 0, 12.5, 4.2 and 4.2%, respectively. The occurrence of >G3 side effects was the most frequent in the MDR1 2677 mutant group at 39%. The disease-free survival and overall survival tended to be longer in the MDR1 2677 mutant group., Conclusions: A correlation between the SNP of MDR1 2677 and drug response in patients receiving GEM chemotherapy.

Background/aims: Since there is a difference in the slice thickness between preoperative images of liver metastases (1-3mm slices) and surgical liver pathology specimens (5mm slices), micrometastases may not be detected in these specimens. In addition, the accuracy of preoperative imaging for the detection of metastases degenerated by chemotherapy is unclear., Methodology: Five patients with liver metastases from colorectal cancer who had received adjuvant chemotherapy and undergone hepatectomy were included. The whole resected liver was sliced at approximately 1mm intervals and the slices were examined carefully for gross lesions. The preoperative CT and EOB-MRI findings of each lesion were compared with gross and histopathological findings., Results: The accuracy of EOB-MRI was higher than that of CT for the detection of liver metastases. The number of lesions detected on EOB-MRI was in agreement with that of histopathologically proven liver metastases in 4 of the 5 patients. All lesions that were grossly identified but turned out to be non-neoplastic were regenerative nodules associated with drug-induced liver injury or lobular nodules associated with marked fatty change, measuring about 1mm in diameter., Conclusions: EOB-MRI was the most accurate method for the preoperative detection of liver metastases, enabling the visualization of almost all liver metastases.

We previously reported that the administration of bevacizumab for pancreatic neuroendocrine tumors inhibited angiogenesis in the host, resulting in tumor growth inhibition. In light of these results, we compared the effect of bevacizumab/gemcitabine/S-1 combination therapy vs. bevacizumab monotherapy. The QGP-1 pancreatic neuroendocrine carcinoma cell line and the BxPC-3 ductal cell carcinoma cell line were transplanted into the subcutaneous tissue of mice, and the mice were treated for 3 weeks with bevacizumab [50 mg/kg intraperitoneally (i.p.) twice weekly], gemcitabine (240 mg/kg i.p. once weekly) and S-1 (10 mg/kg orally five times weekly). The antitumor effect and side effects were evaluated by measuring the tumor volume and weight and by changes in body weight, respectively. The tumor volume became smaller (from the maximum volume) in the group treated with bevacizumab, gemcitabine and S-1 (BGS) and the group treated with bevacizumab and gemcitabine (BG). A significant difference was noted in the tumor weight between the BG group and the group treated with bevacizumab alone. A relatively significant decrease in the body weight was observed in the BGS and BG groups. We conclude that gemcitabine is appropriate as a drug used in combination with bevacizumab for pancreatic neuroendocrine tumors.

In addition to the use of chemotherapeutic agents for the prevention of multiple liver metastases from colorectal cancer, the anti-vascular endothelial growth factor (VEGF) antibody, bevacizumab, is often used, and its effectiveness has been established. By contrast, it has been reported that the use of bevacizumab prior to or following surgery delays wound healing or liver regeneration. In this study, we investigated whether the administration of bevacizumab following hepatectomy inhibits remnant liver regeneration or the growth of remnant metastases. Mice were partially hepatectomized (31% of the liver was removed), transplanted with the murine colorectal cancer cell line, CT26, in the remnant lobe, and intraperitoneally injected with bevacizumab (4 mg/kg) for a total of 6 times. Serum VEGF levels were measured on day 1 following surgery, and each lobe of the liver was weighed on day 14. Serum VEGF levels in non-hepatectomized, tumor-bearing mice exceeded those in their non-tumor-bearing counterparts; however, the administration of bevacizumab did not reduce the serum VEGF levels. The volume of the liver lobe of the hepatectomized, CT26-transplanted and non-CT26-transplanted mice was 1,349.6 and 735.5 mg, respectively, indicating rapid growth of the CT26 transplant (p=0.023). The volume of the CT26-transplanted lobe of the bevacizumab-administered mice was 1,379.0 mg, which was not significantly different from that (1,349.6 mg) of the non-bevacizumab-administered mice. The volume of the remnant lobe of the bevacizumab-administered mice was 1,051.0 mg, which did not significantly differ from that (957.3 mg) of the non-bevacizumab-administered mice. The administration of bevacizumab following hepatectomy did not delay remnant liver regeneration, and did not suppress the growth of metastases in the remnant lobes or remnant liver regeneration.

Background/aims: Anticancer drugs are essential to pancreatic cancer therapy. The multidrug-resistance 1 (MDR1) gene codes for one of the ATP binding cassette (ABC) transporters. The neutralizing antibody of MDR1 reduces the activity of MDR1 and may add to the sensitivity of anti-cancer drugs. We investigated the relationship of the single nucleotide polymorphisms (SNPs), 2677G and 3435C, in the MDR1 gene and the effect of the anti-MDR1 single chain antibody (scAb) using pancreatic cancer cell lines., Methodology: We exposed the pancreatic cancer cell lines, AsCP-1, Panc-1, BxPC-3, MIAPaCa-2 and QGP-1 to 0.1-1,000µ g/mL of 5-FU for 72h and calculated the cytotoxic reactions. Combined therapy with an established anti-MDR1 neutralizing scAb and 10µg/mL of 5-FU was also performed., Results: AsCP-1 contained wild types of MDR1 2677G and 3435C, and showed the most 5-FU resistance. The anticancer effect of AsPC-1 increased with anti-MDR1 scAb, but the effect was not significant compared with other cell lines., Conclusions: The cells with the wild type SNPs of MDR1 showed drug resistance, but we were not able to confirm a remarkable effect of the anti-MDR1 antibody.

The normal pancreas has an abundant blood flow, in contrast to pancreatic cancer, which is a hypovascular tumor. During hypoxia under a hypovascular environment, the transcription factor hypoxia-inducible factor-1α (HIF-1α) is activated. High HIF-1α expression reduces sensitivity to gemcitabine (GEM) which is used as a treatment for pancreatic cancer. The objective of this study was to clarify HIF-1α expression in pancreatic cancer and the association of its effects to GEM treatment. We used the human pancreatic ductal carcinoma cell lines AsPC-1 and BxPC-3 to evaluate cell proliferation, HIF-1α protein expression and sensitivity to GEM in a hypoxic environment of 1% O2 in 48 pancreatic cancer patients who received adjuvant GEM treatment after pancreatectomy. We divided the patients according to HIF-1α expression and the presence of single nucleotide polymorphisms, and we based our evaluation on the adverse events associated with GEM chemotherapy and patient outcome. The hypoxic environment promoted cell proliferation, induced HIF-1α expression and increased GEM resistance, especially in AsPC-1 cells, which included a mutant homozygote for HIF-1α(C1772T). There were no significant differences between the HIF-1α(-) and HIF-1α(+) groups in either adverse events or patient outcomes. HIF-1α enhanced neo-microvascularity in a hypoxic environment and increased drug resistance. The period until recurrence was shorter in the patients with a strong HIF-1α expression, than that in those with a weak HIF-1α expression.

Background/aims: Pancreaticobiliary maljunction (PBM) is a high risk factor in biliary tract cancer. The relation of CD44s and CD44v6 expression in biliary epithelium with PBM and the carcinogenetic process was immunohistochemically examined., Methodology: One hundred and seven lesions were randomly selected from gallbladders and bile ducts, which were resected from 25 patients with PBM, and immunostaining for CD44s, CD44v6 and MIB-1 was carried out., Results: Among gallbladder lesions, cancerous lesions (dysplasia, cancer) had a higher immunoreactivity with statistical significance for CD44s and CD44v6 compared to non-cancerous lesions (normal, hyperplasia). In bile ducts as well, cancerous lesions had a higher immunoreactivity for CD44s and CD44v6. In both gallbladders and bile ducts, positive cases of CD44s and CD44v6 had a higher statistical significance of Ki-67 labeling index in comparison with negative cases., Conclusions: In biliary epithelium with PBM, CD44 was indicated to be strongly related to cancer progression via an increase of cellular proliferative potential.

At present, no effective chemotherapy for pancreatic neuroendocrine carcinoma (PNEC) exists. However, anti-angiogenic therapy is expected to be effective for PNEC, a hypervascular tumor. We treated PNEC and hypovascular pancreatic ductal cell carcinoma (DCC) cell lines with the anti-vascular endothelial growth factor (VEGF) antibody bevacizumab, and compared the antitumor effect between the two different types of cell lines. The PNEC cell line QGP-1 and the DCC cell lines BxPC-3 and AsPC-1 were used. We evaluated the ability of the cell lines to proliferate and secrete VEGF in vitro, the antitumor effect of bevacizumab administration in vivo and the side effects of bevacizumab on the pancreas in a caerulein-induced pancreatitis model. Comparison of the QGP-1 and DCC cell lines showed that QGP-1 secreted a higher level of VEGF under a hypoxic environment than the DCC cell line, and bevacizumab exerted the most marked growth-inhibitory effect on QGP-1; the number of intratumoral blood vessels decreased and the percentage of proliferating cells was approximately the same. In the pancreatitis model, bevacizumab administration did not adversely affect the pancreatitis or the associated hypoxic environment. Bevacizumab does not affect the pancreas itself; therefore, its potent inhibitory effect on the growth of pancreatic neuroendocrine tumors alone can be expected.

Background: Recently, the usefulness of endoscopic ultrasound (EUS) elastography has been reported for the diagnosis of pancreatic lesions. In the present study, we retrospectively assessed EUS elastography as a diagnostic tool by evaluating tissue elasticity distribution and elasticity semiquantification, using the strain ratio (SR) of tissue elasticity, in patients with pancreatic masses., Methods: One hundred and nine patients who underwent EUS elastography between September 2006 and May 2009 were retrospectively evaluated. The final diagnosis was chronic pancreatitis (CP) in 20 patients [6 with non-mass-forming pancreatitis, 7 with mass-forming pancreatitis (MFP), and 7 with autoimmune pancreatitis (AIP)], pancreatic cancer (PC) in 72, pancreatic neuroendocrine tumor (PNET) in 9, and normal pancreas in 8. The tissue elasticity distribution calculation was performed in real time, and the results were represented in color in fundamental B-mode imaging. In addition, we performed quantification using the SR (non-mass area/mass area)., Results: Elastography for all PC patients showed intense blue coloration, indicating malignant lesions. In contrast, MFP presented with a mixed coloration pattern of green, yellow, and low-intensity blue. Normal controls showed an even distribution of green to red. The mean SR was 23.66 ± 12.65 for MFP and 39.08 ± 20.54 for PC (P < 0.05)., Conclusions: Endoscopic ultrasound elastography is a promising diagnostic tool for defining the tissue characteristics of pancreatic masses. In addition, semiquantitative analysis of elasticity using the SR may allow the differentiation of MFP from PC.

Background and Purpose: Recently, biodegradable pancreatic stents have been designed and placed in vivo and in vitro. The aim of this study is to investigate the feasibility of endoscopic stenting using the novel, braided, self-expandable, biodegradable, pancreatic and biliary stent in a pig model., Methods: A braided, self-expandable, biodegradable stent was endoscopically placed into the pancreatic duct and bile duct in 4 pigs. Eventually, necropsy was performed to evaluate the stent placement after the procedure., Results: Pancreatic and biliary stents were successfully inserted and easily deployed across the papilla into the main pancreatic duct and bile duct, respectively, in all the animals under endoscopic and fluoroscopic guidance. Necropsy performed immediate following stent placement, found that stents had been placed across the papilla and stent expansion had occurred in all cases., Conclusions: Although this stent is not radiopaque and the number of cases was small in this experimental study, endoscopic stenting using this novel, braided, self-expandable, biodegradable, pancreatic and biliary stent was feasible in the pig model. Further animal studies to evaluate the short-term patency, tissue reactivity and degradability of the stents are warranted.

Purpose: For hepatic tumors that cannot be identified on routine ultrasonography (US), we marked the target area using real-time virtual sonography (RVS) with indocyanine green (ICG)-ethanol (1:100) during surgery, and performed hepatic resection while observing the fluorescence., Experiment: An ICG-ethanol mixture locally injected into mouse liver was retained in the same area for more than 4 h. The same mixture locally injected into pig liver at a depth of 3 cm could be observed using an infrared camera., Case: An ICG-ethanol mixture (500 μl) was locally injected under RVS guidance into a metastatic hepatic tumor that was visible only on magnetic resonance imaging (MRI), and hepatic resection was performed while Photodynamic Eye (PDE) images were being observed. The metastatic lesion (3 mm in diameter in a pathological specimen) could be successfully resected., Conclusion: This method was useful for resecting US-invisible hepatic tumors.

Background: Magnetic compression anastomosis (MCA) is a revolutionary, minimally invasive method of performing choledochoenterostomy or choledochocholedochostomy without using surgical techniques in patients with biliary stricture or obstruction. Herein, we describe a case series of MCA for severe biliary stricture or obstruction, which could not be treated with conventional therapies., Patients and Methods: Two patients with biliary obstruction were treated using MCA for choledochocholedochostomy and choledochoenterostomy at Tokyo Medical University Hospital and Tokyo Medical University Hachioji Medical Center. Endoscopically, a samarium-cobalt (Sm-Co) rare-earth magnet was placed at the superior site of obstruction through the percutaneous transhepatic biliary drainage route and another Sm-Co magnet was placed at the inferior site of obstruction. A comprehensive computer-aided literature search for MCA was performed up to September 2009 by using MEDLINE and EMBASE., Results: MCA techniques enabled complete anastomosis in both cases without procedure-related complications., Conclusions: The MCA technique is a revolutionary method of performing choledochocholedochostomy and choledochoenterostomy interventionally in patients with biliary obstruction, for whom the conventional endoscopic procedure is not available, or in candidates who are deemed unsuitable for surgery.

Background/aims: Pancreaticobiliary maljunction (PBM) is a high risk factor of biliary tract cancer. The chemopreventive effects of Vitamin K2 (menaquinone-4: MK4) in a hamster PBM model were investigated., Methodology: The extrahepatic bile duct at the distal end of the common duct was ligated and cholecystoduodenostomy was performed (Group I). The same surgery was performed and from 4 weeks after surgery, 10 mg/kg of N-nitrosobis (2-oxopropyl) amine was subcutaneously injected once a week with a one-week interval (Group II). In addition of Group II, MK4 was orally administered once a day, five times with every week (Group III). The hamsters were sacrificed 20 weeks after surgery and histopathological findings of gallbladder were investigated., Results: Group I showed predominantly proper epithelium without cancer. In Group II, atypical epithelium (AE) was observed in 75% of animals and early cancer was observed in 25%. Group III showed less AE and no cancer. The PCNA labeling index in Group III was statistically significantly lower than in Group II. In addition, no statistically significant differences were noted among the groups in terms of the apoptosis labeling index., Conclusions: MK4 suppressed biliary carcinogenesis by the induction of cell cycle arrest in a hamster biliary carcinogenetic model.

We describe the surgical method of cases showing a distended gallbladder. Because the most important thing does not cause biliary tract injury, it is to find orientation carefully. The frequency of incidental gallbladder cancer was in 7 (0.7%) of the 983. Only cholecystectomy is necessary to be performed for Tis or T1 cancer, and surgery has to be changed to radical surgery for T2 cancer or deeper invasion. Laparoscopic cholecystectomy is already an established standard operation. In the presence of acute or severe chronic inflammation, special attention should be paid to these points.

Magnetic compression anastomosis (MCA) is a minimally invasive method of performing choledochocholedochostomy without surgery in patients with biliary stricture or obstruction. We describe a successful case involving magnetic compression duct-to-duct biliary reconstruction in right-lobe living donor liver transplantation (RL-LDLT). Endoscopically, a samarium-cobalt (Sm-Co) rare-earth magnet was placed at the superior site of obstruction via the percutaneous transhepatic biliary drainage route, and another Sm-Co magnet was also placed at the inferior site of obstruction with the aid of an endoscope. MCA techniques enabled complete anastomosis without procedure-related complications. In conclusion, the MCA technique is a revolutionary method of performing choledochocholedochostomy in patients with biliary obstruction after LDLT.

A novel technique of anastomosis with a short-term degradable stent for pancreaticojejunostomy has been developed and its shape-retentive capability evaluated under conditions of exposure to digestive fluid and its clinical feasibility. The stent was braided manually using Monocryl and PDS-II of 2-0 wire and heat treated at 80 degrees C for 3 hours under a vacuum. We measured its maximum resistance to compression strength and elastic modules of the stent. We evaluated the feasibility of the stent in 8 patients under pancreaticojejunostomy and hepaticojejunostomy. The compression strength of the stent was preserved for about 3 weeks, after which point it collapsed. The elastic features were retained under exposure to gastric fluid, but they were lost under an intestinal juice exposure. Using this stent, "suturing of tissue with the stent on" was possible as a novel technique of anastomosis and induced few complications.

The aim of the present study was to determine whether symptoms of atrial fibrillation (AF) differ between patients with and without subsequent permanent AF. Sixty-four patients (68 +/- 10 years old, 45 males) were recruited. AF follow-up was started at the age of 61 +/- 10 years and accomplished in a median period of 4.9 years (396 person-years). Permanent AF, defined as lasting > 180 days, developed in 17 patients (14 males) (43 per 1000 person-years). The AF follow-up period was longer in the permanent AF group than in the non-permanent AF group (median, 9.8 versus 4.2 years, P < 0.001). For baseline characteristics, hypertension was less frequent in the permanent AF group than in the nonpermanent AF group (18% versus 45%, P < 0.05). A retrospective questionnaire survey regarding initial AF symptoms was conducted. The severity of AF symptoms by a 4-grade scale was significantly milder in the permanent AF group than in the nonpermanent AF group (P < 0.05). Cox proportional hazards model analysis revealed that the severity of initial AF symptoms was related to the subsequent development of permanent AF (hazard ratio 0.46 per grade, 95% confidence interval 0.23 - 0.93, P < 0.05), but age, gender, hypertension, diabetes mellitus, organic heart disease, and left atrial dimension were not. The permanent AF-free rate was significantly lower in 33 patients with mild symptoms than in 31 patients with severe symptoms (log-rank test, P < 0.05). These results point to an inconspicuous feature in the development of permanent AF.

Background: Recently, a second-generation photosensory agent for photodynamic therapy (PDT), mono-L: -aspartyl chlorine e6 (NPe6), which degrades rapidly in vivo, has been developed. We evaluated its feasibility and efficacy for treatment in biliary tract carcinoma., Methods: A transmittance of semiconductor laser light (664 nm), sensitivity of a human biliary tract carcinoma cell line, and disorder to normal tissue including Glissonian constructs and adjacent hepatocytes were investigated., Results: The transmittance of the laser was 85-91% through yellow clear bile and that of the bile including 50 microg/ml NPe6 was 17-48%. The effective concentration of NPe6 which showed LD50 for a cell line was 12.5 microg/ml, and that of LD95 was 25 microg/ml. NPe6 in the supernatant reduced laser transmissiveness, but it had little influence on the antitumor effect in supernatant with or without NPe6. The NOZ cell-tumor volume was reduced significantly 14 days after irradiation in the PDT group (PDT 69.9 +/- 44.6 mm(3) vs control 296.3 +/- 239.9 mm(3) P < 0.05). No severe hepatic disorder including Glisson components was observed by the histological findings., Conclusion: NPe6 PDT was effective in carcinomas even in the presence of bile, and causes no serious complication for the liver and Glisson structure. Therefore, NPe6 PDT will be a useful candidate as a new therapy for biliary tract carcinomas.

Aim: To examine the coexistence of metachronous and synchronous cancer in branch duct intraductal papillary mucinous neoplasms of the pancreas (IPMN)., Methods: We reviewed the records of 145 patients with branch duct IPMN between January 1991 and April 2008 and assessed the relationship between IPMN and intra- or extra-pancreatic carcinoma and the outcome of IPMN., Results: The mean observation period was 55.9 +/- 45.3 mo. Among the 145 patients, the frequency of extra-pancreatic cancer was 29.0%. The frequency of gastric cancer, colon cancer, breast cancer, and pancreatic cancer were 25.5%, 15.7%, 13.7%, and 9.8%, respectively. Twenty (13.8%) of the patients died. The cause of death was extra-pancreatic carcinoma in 40%, pancreatic cancer in 25%, IPMN per se in 20%, and benign disease in 15% of the patients., Conclusion: The prognosis for IPMN depends not on the IPMN per se, but on the presence of intra- or extra-pancreatic cancer.

Humanized antibodies vary and have certain effects, but they are expensive and require repeated administration. We developed cells which constantly express a humanized antibody, and we performed anticancer humanized antibody therapy involving cell transplantation. Genes with the same amino acid sequence as that of the variable region of trastuzumab (Herceptin) as the humanized anti-HER2 monoclonal antibody were produced by overlap-PCR and were connected to the anti-human CD16 antibody [anti-HER2+anti-CD16 single-chain antibody (anti-HER2+CD16 scAb)]. For transplantation, stem cell-like cells that are immunologically tolerant and do not transform into cancer [mouse embryo fibroblast cell line C3H10T1/2 (10T1/2)] were used. The antibody was incorporated into 10T1/2 (antibody-expressing cells) using the pMX-IRES-EGFP retroviral vector. Cell supernatants and human monocytes were exposed to the human breast cancer strain HTB131 expressing HER2, and the in vitro antibody-dependent cell-mediated cytotoxicity (ADCC) effects were evaluated. After the transplantation of antibody-expressing and HTB131 cells into SCID mice, human monocytes were intermittently administered, and the in vivo ADCC effects were evaluated. We found that the ex vivo dead cell rate was 15.4% for Herceptin, 5.6% for anti-HER2+CD16 scAb, 1.5% for anti-CD16 scAb, and 2.1% for the control, demonstrating the antitumor effects of anti-HER2+CD16 scAb. In an antibody-expressing cell transplantation model, the inhibitory effects of this antibody on HTB131 cell establishment were observed. In conclusion, the establishment of breast cancer cells in the peritoneum was inhibited by the transplantation of antibody-expressing cells. Since this method requires cell transplantation only once, the drug cost may be reduced.

The aim of this study is to evaluate a prototype variable stiffness duodenoscope (VSD) for diagnostic and therapeutic ERCP in comparison with standard duodenoscopes. We performed retrospective analysis on the success rate of intubation of the second duodenum, overall procedural success rate, and comparative frequency of the necessity to change duodenoscopes from standard JF-260V and TJF 260V or to change stiffness using the VSD. A total of 213 nonconsecutive procedures in 196 patients with pancreaticobiliary diseases. There was no statistically significant difference in endoscope intubation rate or technical success rate between the different duodenoscopes. In one patient with severe duodenal stenosis, the VSD using the moderately stiff mode allowed the major papilla to be reached when the TJF-260V endoscope could not. There were no serious procedure-related adverse events. In conclusion, while the VSD performed well, the present models do not appear to offer obvious advantages over the standard duodenoscopes for routine diagnostic and therapeutic ERCP. Prospective studies may be warranted to identify those patients who would benefit from this new technology.

Inhibition of mitochondrial permeability transition (mPT) has emerged as a promising approach for neuroprotection and development of well-tolerated mPT inhibitors with favorable blood-brain barrier penetration is highly warranted. In a recent study, 28 clinically available drugs with a common heterocyclic structure were identified as mPT inhibitors e.g. trifluoperazine, promethazine and nortriptyline. In addition, neuroprotection by structurally unrelated drugs e.g. neurosteroids, 4-hydroxy-tamoxifen and trimetazidine has been attributed to direct inhibition of mPT. The regulation of mPT is complex and highly dependent on the prevailing experimental conditions. Several features of mPT, such as swelling, depolarization or NADH oxidation, can also occur independently of the mPT phenomenon. Here, in isolated rodent brain-derived and human liver mitochondria, we re-evaluate drugs promoted as potent mPT inhibitors. We address the definition of an mPT inhibitor and present strategies to reliably detect mPT inhibition in vitro. Surprisingly, none of the 12 compounds tested displayed convincing mPT inhibition or effects comparable to cyclophilin D inhibition by the non-immunosuppressive cyclophilin inhibitor D-MeAla(3)-EtVal(4)-Cyclosporin (Debio 025). Propofol and 2-aminoethoxydiphenyl borate (2-APB) inhibited swelling in de-energized mitochondria but did not increase calcium retention capacity (CRC). Progesterone, trifluoperazine, allopregnanolone and 4-hydroxy-tamoxifen dose-dependently reduced CRC and respiratory control and were thus toxic rather than beneficial to mitochondrial function. Interestingly, topiramate increased CRC at high concentrations likely by a mechanism separate from direct mPT inhibition. We conclude that a clinically relevant mPT inhibitor should have a mitochondrial target and increase mitochondrial calcium retention at concentrations which can be translated to human use.

Pancreaticobiliary maljunction (PBM) is associated with the occurrence of biliary cancer due to pancreatobiliary reflux. We present a case of simultaneous double cancer of the gallbladder and bile duct. A 77-year-old woman who had jaundice, intra- and extra-hepatic biliary ductal dilatation and a space-occupying lesion in the gallbladder and lower bile duct underwent pancreatoduodenectomy. The gallbladder cancer showed papillary carcinoma without mutation of the K-ras gene and with p53 non-sense mutation of CCA (Pro) to CA (Stop) on codon 301 in exon 8. The bile duct cancer revealed a well-differentiated adenocarcinoma without mutation of the K-ras gene and with p53 miss-sense mutation of GTG (Val) to GAG (Glu) on codon 272 in exon 8. There were no mutations of either the K-ras or p53 gene in non-cancerous epithelia. In contrast, only the mucosa of the common channel had p53 protein accumulation and high cell proliferation activity. Therefore, the genetic pathway might be the same in both the gallbladder and bile duct cancer, and a high potential for carcinogenesis might be present in the epithelium of the common channel in patients with PBM.

Aim: This study was designed to clarify differences in serum phospholipid fatty acid compositions and estimated desaturase activities between Japanese men with and without metabolic syndrome (MetS)., Methods: From among 227 males, 40 to 59 years of age, excluding those receiving treatment for lipid disorders, 165 subjects (including 27 with MetS) were selected for this study. Serum phospholipid fatty acid compositions were determined, and desaturase activities were estimated., Results: The C15:0 and C17:0 fatty acids associated with hepatic function were lower, while the C20:3n-6 and C20:4n-3 fatty acids were higher, in subjects with than without MetS (p<0.05). The estimated desaturase activity for D5D(n-6) was lower in subjects with than without MetS (p<0.01). Body fat percentage was an independent negative predictor of C17:0, and a positive predictor of log C20:3n-6 and log C20:4n-3 (p<0.01). HDL-C was an independent negative predictor of log C15:0 and of C17:0 (p<0.01)., Conclusion: Decreases in minor saturated fatty acids, accumulation of C20:3n-6 and C20:4n-3 and low estimated D5D activity were confirmed to be associated with MetS.

A case of gliosarcoma composed of glioblastoma and liposarcoma is presented. A 70-year-old Japanese man was admitted to hospital because of dysarthria and aphasia. Magnetic resonance imaging indicated a brain tumor located in the temporal-parietal area of the left hemisphere. He rejected any therapy and died of respiratory failure. At autopsy the tumor was well-demarcated with firm consistency and myxoid appearance, accompanied by necrosis and hemorrhage. Microscopically the tumor consisted of both glial and sarcomatous components, compatible with a gliosarcoma. Lipoblast-like tumor cells were identified in the sarcomatous area. Glial component was observed in the periphery and was diffusely positive for CD56 and S100 protein and focally for glial fibrillary acidic protein. Only a small number of tumor cells in the sarcomatous area expressed neurogenic markers. Lipoblast-like tumor cells were positive for S100 protein but negative for any other neurogenic markers. A significant number of tumor cells were positive for retinoblastoma protein (pRB) in the glial area, whereas only a few of them were positive in the sarcomatous area, indicating alteration of pRB in sarcomatous component. The present tumor is a rare gliosarcoma with liposarcomatous differentiation; alteration of pRB may play a role in sarcomatous transformation of glial component.

The crystalline structure dependence of enzymatic degradation behavior was investigated for the polymorphic poly(3-hydroxypropionate) (P3HP), which has a basic backbone chemical structure of bacterial poly(3-hydroxyalkanoate)s (P3HAs). The P3HP films consisting of the beta-, gamma-, and/or delta-form crystal were cast or melt-crystallized as reported previously (Macromolecules 2005, 38, 6455; Macromolecules 2006, 39, 194-203) by controlling the molecular weight, crystallization temperature, and/or temperature of the melt. Their thermal properties, crystalline structures, morphologies, and (13)C solid spin-lattice relaxation dynamics were characterized by the differential scanning calorimetry, the wide-angle X-ray diffraction, the small-angle X-ray scattering (SAXS), and the (13)C solid-state NMR spectra (SNMR), respectively. Both the crystallinities and the lamellar thicknesses of P3HP films were found to decrease roughly in the order of beta-form > (or approximately) gamma-form > delta-form. From previous work, which indicates that the P3HA enzymatic degradation depends only on the degree of crystallinity and the lamellar thickness, their enzymatic degradation rates are then expected to increase in the order of beta-form < (or approximately) gamma-form < delta-form. Unexpectedly, their experimental P3HP enzymatic degradation rates in the presence of P3HA depolymerase isolated from Ralstonia pickettii T1 increase in the reverse order, i.e., delta-form < gamma-form < beta-form. The weight loss rate of the delta-form film is almost 1 order of magnitude smaller than that of the fastest degraded beta-form film. It is then strongly indicated that the crystalline structure plays a strikingly decisive role in the enzymatic degradation of P3HP. In particular, only when the conformation of crystalline chain accords with that of the bacterial poly(3-hydroxybutyrate) (P3HB) sample, i.e., the 2 1 helix conformation, is the P3HP sample degraded as slow as the P3HB sample. The inherent reason responsible for the unique P3HP enzymatic degradation behavior has been further clarified by comparing the molecular interaction and dynamics of polymorphic P3HP crystals.

Background/purpose: It has been reported that circulating tumor cells (CTCs) can be used to predict survival in metastatic breast cancer. In this preliminary study, we examined the level of CTCs in pancreatic cancer (PC) patients to elucidate whether we could predict survival in PC., Methods: The eligible subjects, at Tokyo Medical University Hospital, were 26 patients with PC, 11 with chronic pancreatitis, and 10 healthy volunteers. Three PC patients underwent surgery, 18 patients (who were stage IV) were treated with gemcitabine (GEM), and 5 patients received best supportive care (BSC)., Results: The CTC count was 1/7.5 ml blood or higher (defined as positive) in 11 of the 26 patients (42%; mean, 16.9/7.5 ml blood; range, 1-105/7.5 ml blood). Gemcitabine was administered to 6 of the 11 CTC-positive patients (3.8 courses on average). The treatment was continued for more than three courses in 2 patients, in both of whom the CTC count was only 1/7.5 ml blood. Operation was performed in 1 of the 11 CTC-positive patients. The remaining 4 patients of the 11 CTC-positive patients received only BSC. CTC was negative in 15 patients with PC (stage II, 1; stage III, 1; stage IVa, 7; and stage IVb, 6) and in the subjects with benign conditions. The median survival times (MSTs) of the CTC-positive and-negative patients were 110.5 and 375.8 days (P < 0.001). When the analysis was limited to the 14 stage-IVb patients, the MSTs of the CTC-positive and-negative patients were 52.5 and 308.3 days (P < 0.01)., Conclusions: The present study demonstrated that the detection of CTCs in peripheral blood may be useful to predict prognosis in patients with PC.

Background/purpose: The VIO soft-coagulation system (SC) is a new device for tissue coagulation. We hypothesized that this device would be an effective tool for sealing small pancreatic ducts, thus reducing pancreatic fistula following pancreatectomy., Methods: To confirm whether the SC could be used to seal small pancreatic ducts, we measured the burst pressure in sealed ducts in mongrel dogs. Eight dogs underwent distal pancreatectomy, with the remnant stump coagulated by using the SC. The animals were necropsied on postoperative day 10. In a clinical trial, 11 patients who underwent pancreatoduodenectomy with SC treatment (SC group), and 24 patients who underwent pancreatoduodenectomy without SC treatment (non-SC group) were compared., Results: In the experimental study, the burst-pressure test revealed that the SC had efficiently sealed the small pancreatic ducts. Histological examination revealed completely obstructed pancreatic ductal structures, ranging from large pancreatic ducts (diameter, 500 microm) to microscopic ducts. No pancreatic leakage was observed following distal pancreatectomy without main pancreatic duct (MPD) suturing in dogs that had an MPD diameter of less than 500 microm. In the clinical trial, pancreatic fistula developed in only one patient (9.1%) in the SC group, but a pancreatic fistula developed in five patients (20.8%) in the non-SC group., Conclusions: This novel technique using the SC is an effective procedure for preventing the development of pancreatic fistula following pancreatectomy.

Reaction processes of poly[(R)-3-hydroxybutyric acid] (P(3HB)) with two types of poly(hydroxybutyric acid) (PHB) depolymerases secreted from Ralstonia pickettii T1 and Penicillium funiculosum were characterized by means of atomic force microscopy (AFM) and quartz crystal microbalance (QCM). The PHB depolymerase from R. pickettii T1 consists of catalytic, linker, and substrate-binding domains, whereas the one from P. funiculosum lacks a substrate-binding domain. We succeeded in observing the adsorption of single molecules of the PHB depolymerase from R. pickettii T1 onto P(3HB) single crystals and the degradation of the single crystals in a phosphate buffer solution at 37 degrees C by real-time AFM. On the contrary, the enzyme molecule from P. funiculosum was hardly observed at the surface of P(3HB) single crystals by real-time AFM, even though the enzymatic degradation of the single crystals was surely progressed. On the basis of the AFM observations in air of the P(3HB) single crystals after the enzymatic treatments, however, not only the PHB depolymerase from R. pickettii T1 but also that from P. funiculosum adsorbed onto the surface of P(3HB) crystals, and both concentrations of the enzymes on the surface were nearly identical. This means both enzymes were adsorbed onto the surface of P(3HB) single crystals. Moreover, QCM measurements clarified quantitatively the differences in detachment behavior between two types of PHB depolymerases, namely the enzyme from R. pickettii T1 was hardly detached but the enzyme from P. funiculosum was released easily from the surface of P(3HB) crystals under an aqueous condition.

Background: Gemcitabine is an efficacious cytotoxic agent used in the treatment of unresectable pancreatic carcinoma (PC). Recently, gemcitabine resistance has been associated with the ribonucleotide reductase subunit M2 (RRM2). In this prospective study, we hypothesized that RRM2 expression in PC biopsy specimens would be a significant predictor of outcome., Methods: RRM2 mRNA expression in 35 endoscopic ultrasonography-guided fine needle aspiration biopsy (EUS-FNAB) samples was quantified using real-time quantitative reverse transcription-polymerase chain reaction., Results: Thirty-one of 35 biopsy specimens could be assessed for RRM2 expression levels. The mean RRM2 expression relative to glyceraldehyde-3-phosphate dehydrogenase was 0.248 (range, 0.00739 to 0.858). Eighteen patients (64.5%) had low RRM2 levels, and 13 patients (35.5%) had high RRM2 levels with a cutoff of 0.1. The median survival was 8.8 months for patients with low RRM2 levels and 5.0 months for patients with high levels (P < 0.05). In the low RRM2 expression group, a complete response (CR) was observed in one patient, and a partial response (PR) was observed in eight patients. In contrast, in the high RRM2 expression group, PR was observed in one patient, and CR was not observed. The overall response rate between the high and low expression groups was significantly different (50.0% vs. 7.7%, P < 0.05)., Conclusions: RRM2 mRNA expression of EUS-FNAB specimens is a key predictive marker of survival in gemcitabine-treated patients with PC.