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Emphysematous cholecystitis (EC) is a severe and rare variant of acute cholecystitis characterized by ischemia of the gallbladder wall with gas-forming bacterial proliferation. Open cholecystectomy is traditionally the gold standard approach to treatment due to difficulty in isolating Calot’s triangle in the setting of intense inflammation. We present a case of EC successfully and safely treated by laparoscopic surgery.

Highlights • Isolated spontaneous dissection of the superior mesenteric artery is very rare condition. • Imaging studies are effective for diagnosis. • Common treatment strategy consists of three methods as follows; conservative therapy, endovascular treatment, and surgery. • The etiology and the best treatment have not been established yet., Introduction Isolated spontaneous dissection of the superior mesenteric artery (SMA) is rare and a treatment strategy has not been established yet. In this paper, we present our experience with two cases and review the literature. Presentation of case Both cases were treated conservatively as they did not show signs of bowel ischemia. They were symptom free with no evidence of disease progression after a median follow-up of 3.5 years. Discussion There are three methods for the treatment of isolated SMA dissection; observation with medical therapy, endovascular surgery, and open surgery. Most patients with isolated SMA dissection can be treated with observation alone. Although the indications for surgery are still controversial, patients with bowel ischemia should undergo invasive treatment in the form of either endovascular or open surgery. Conclusion We recommend observation with medical therapy as the first choice for isolated SMA dissection. However, long term follow-up is necessary as the extent of the dissection may change over time.

Because angiogenesis is essential for tumor growth and metastasis, the development of antiangiogenic agents is an urgent issue in cancer treatment. Zerumbone, a component of subtropical ginger, has been shown to exhibit anticancer activities in various cancer cells; however, little is known about its biological mechanisms against angiogenesis in pancreatic cancer (PaCa). Here, we evaluated the effects of zerumbone on PaCa angiogenesis.The cytotoxicity of zerumbone in PaCa was measured using premix WST-1 cell proliferation assays. The influence of zerumbone on the angiogenic factors vascular endothelial growth factor and interleukin 8 was measured using the reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assays. Changes in nuclear factor-κB (NF-κB) activities were measured using NF-κB transcription factor assays. We also examined the effects of zerumbone on PaCa-induced angiogenesis using angiogenesis assays.Zerumbone inhibited mRNA expression and protein secretion of angiogenic factors and NF-κB activity. Tube formation in human umbilical vein endothelial cells was enhanced by coculture with PaCa cells, and these enhancements were significantly inhibited by zerumbone treatment.Zerumbone blocked the PaCa-associated angiogenesis through the inhibition of NF-κB and NF-κB-dependent proangiogenic gene products.

Foreign body retrieval surgery sometimes does not need excision and reconstruction of the gastrointestinal tract. To avoid unnecessarily large incisions is now feasible for such retrieval surgery. We report herein a case of a patient who underwent retrieval surgery with a single-incision laparoscopic surgery (SILS) to minimize surgical incision. Our case concerns a 72-year-old female with a 3 cm length needle incidentally discovered as a peritoneal foreign body without any apparent symptoms. Retrieval was performed safely with SILS port inserted through a 2.5 cm transverse incision below the umbilicus. Two flexible forceps introduced through the SILS port offer easy operation for retrieving foreign bodies even if dissection or other surgical maneuvers are required. Our technique of abdominal foreign body retrieval with the SILS port is a useful clinical application of minimally invasive surgery for such cases.

Objective Cardiac tamponade is a serious complication of central venous catheter (CVC) insertion. The position of the carina has been shown to be near the pericardial reflection and can easily be identified as a landmark on routine chest x-ray (CXR). The purpose of this study was to show a simple way to predict optimal CVC depth, thereby facilitating safe positioning of the CVC tip. Methods Subjects included 119 inpatients undergoing gastroenterologic surgery. Central venous catheterization was performed through the right internal jugular vein or the right subclavian vein. The insertion depth was measured. Postoperatively, the CVC tip position was confirmed by CXR and the distance between the CVC tip and the carina was measured. We compared the “original measurement” up to the carina from the insertion point with the “calculated measurement” derived by adding half the length of the right clavicle and the vertical length between the sternal head of the right clavicle and the carina on the CXR. Results There was a significant correlation between the original measurement and the calculated measurement when performed through the internal jugular vein and the subclavian vein. Conclusion The appropriate length of CVC inserted through the right internal jugular vein or right subclavian vein could be estimated by the calculated measurement of adding half the length of the right clavicle and the vertical length between the sternal head of the right clavicle and the carina.

Background and Objectives Interleukin (IL)-1α plays an important role in colon cancer progression and angiogenesis. We here asked whether IL-1α derived from cancer cells modulates vascular endothelial cell growth, migration and tubule formation. Methods The existence of IL-1α mRNA and protein in colon cancer cell lines (WiDr, HT-29, Caco-2, COLO 320) were investigated with RT-PCR and ELISA. Proliferation and invasion were investigated by MTS assay and Matrigel-double chamber assay. To answer our main question, we performed angiogenesis assay used an in vitro model consisting of co-cultivated tumor cells and stromal cells. Results IL-1α mRNA and protein were detected in highly metastatic colon cancer cells (WiDr and HT-29). Recombinant IL-1α significantly enhanced growth and invasiveness of human umbilical vein endothelial cells (HUVEC) (P

CXC-chemokines are involved in the chemotaxis of neutrophils, lymphocytes and monocytes. However, role of these chemokines in tumorigenesis, especially with regard to interaction between tumor and its microenvironment, has not been clearly elucidated. The purpose of this study was to analyze the co-operative role of CXCL8 and CXCL12 in the tumor-stromal interaction in pancreatic cancer (PaCa). Using enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR), we initially confirmed the expression of ligands and receptors, respectively, of CXC-chemokines in PaCa and stromal cells. We examined the co-operative role of CXCL8 and CXCL12 in proliferation/invasion of PaCa and human umbilical vein endothelial cells (HUVECs), and in HUVEC tube-formations through tumor-stromal interaction by MTS, Matrigel invasion, and angiogenesis assays, respectively. We detected expression of CXCR4, but not CXCR2, in all PaCa cells and fibroblasts. PaCa cells secreted CXCL8, and fibroblast cells secreted CXCL12. CXCL8 production in PaCa was significantly enhanced by CXCL12, and CXCL12 production in fibroblasts was significantly enhanced by co-culturing with PaCa. CXCL8 enhanced proliferation/invasion of HUVECs but did not promote proliferation/invasion of PaCa. Both recombinant and PaCa-derived CXCL8 enhanced tube formation of HUVECs that were co-cultured with fibroblast cells. CXCL12 enhanced the proliferation/invasion of HUVECs and the invasion of PaCa cells but had no effect on tube formation of HUVEC. We showed that PaCa-derived CXCL8 and fibroblast-derived CXCL12 cooperatively induced angiogenesis in vitro by promoting HUVEC proliferation, invasion, and tube formation. Thus, corresponding receptors CXCR2 and CXCR4 are potential antiangiogenic and antimetastatic therapeutic targets in PaCa.

Epidemiologic studies suggest that fish oil, rich in n-3 polyunsaturated fatty acids (PUFA), possesses antitumor activity, whereas n-6 PUFAs may stimulate the development of cancers. The aim of this study was to evaluate the effects of n-6 and n-3 PUFAs on the growth of pancreatic cancer.The n-6 PUFA arachidonic acid (AA) stimulated the growth of cyclooxygenase (COX) 2 positive human pancreatic cancer (PaCa) cells, which was mediated by COX-2 generated prostaglandin E2 (PGE2) binding to EP2 and EP4 receptors. In contrast, the n-3 PUFA eicosapentaenoic acid decreased the growth of COX-2-positive and COX-2-negative PaCa cells. The COX-2-dependent mechanism of eicosapentaenoic acid was mediated by binding of PGE3 to EP2 and EP4 receptors. Dietary intake of n-3 PUFAs decreased the growth of pancreatic cancers in a xenograft model, which was accompanied by a decrease of PGE2 and an increase of PGE3 in the tumors.Our studies provide evidence that n-3 PUFAs possess antitumor activities, whereas n-6 PUFAs stimulate pancreatic tumor growth. The opposite effects of n-3 and n-6 PUFAs are mediated by the formation of different prostaglandin species. n-3 PUFAs may prove beneficial as monotherapy or combination therapy with standard chemotherapeutic agents in pancreatic cancer patients.

In this study, we examined the role of c-kit receptor (KIT) signal transduction on the proliferation and invasion of colorectal cancer cells. We found that c-kit was expressed in 2 colorectal cancer cell lines as determined by RT-PCR, Western blot, and flow cytometry. In KIT-positive lines, KIT was activated by stem cell factor (SCF). SCF enhanced cellular proliferation of positive lines as demonstrated by the WST-1 proliferation assay. Furthermore, SCF enhanced the invasive ability of KIT-positive cell lines. SCF stimulation upregulated p44/42 mitogen-activated protein kinase (MAPK) and Akt as shown by Western blot. We examined the roles played by p44/42 MAPK and phosphatidylinositol 3-kinase (PI3K)/Akt pathways in proliferation and invasion. PI3K/Akt activity strongly correlated with proliferation and invasion and p44/42 MAPK was correlated with only invasion. In conclusion, the SCF-enhanced proliferation and invasion of KIT-positive colorectal cancer cells is achieved mainly through the PI3K/Akt pathway.

Background The glial cell line–derived neurotrophic factor (GDNF) is a member of neurotrophic polypeptide family, which promotes survival and rescue of various neural cells in the central and peripheral nerve systems. We previously reported that GDNF promotes tumor cell invasion in pancreatic cancer cell lines. The purpose of this study was to investigate GDNF family expression and the status of related receptors in actual cancer tissues, and assess correlations with clinicopathologic behavior. Methods Immunohistochemical assessment of GDNF, neurturin, persephin, artemin, GDNF family receptor α-1 and α-2, and RET was performed for 51 cases of surgically resected pancreatic cancer. Results In all intrapancreatic nerves, GDNF and artermin were expressed strongly. In pancreatic cancer tissues. The expression of RET was stronger than that seen in normal ductal cells and was significantly related to the survival rate after resection ( P = .026) and lymphatic invasion ( P = .014). Intrapancreatic neural invasion was significantly related to the expression of GDNF ( P = .047). Conclusions We conclude that the expression of RET in pancreatic cancer tissues may be a useful prognostic marker and GDNF may play an important role in neural invasion.

Objective Misplacement of subclavian vein catheters has been reported. We assessed the rate of misplacement of a subclavian vein catheter with and without a novel stylet. Methods A prospective, randomized, controlled clinical trial was conducted between September 2001 and June 2003 in a university hospital. Two hundred sixteen adult patients were enrolled to receive subclavian vein catheterization under non-emergency conditions. Patients were randomly assigned to undergo right subclavian vein catheterization with the stylet ( n = 109, stylet group) or without the stylet ( n = 107, control group). The rate of subclavian vein catheter misplacement was determined and risk factors for failure and complications were analyzed. Results There was no incidence of catheter misplacement in the stylet group, but this occurred in 12 patients in the control group in whom the catheter was misplaced into the ipsilateral internal jugular vein (0% versus 11.2%, P = 0.0003). In multivariate analyses, use of the stylet (odds ratio = 0.062, 95% confidence interval = 0.008 to 0.495, P = 0.009) and a close to average body mass index (odds ratio = 0.73, 95% confidence interval = 0.54 to 0.98, P = 0.038) were associated with low risks of complications and failure, respectively. Conclusions This stylet is useful for decreasing the incidence of misplacement of subclavian vein catheters. Body mass index is predictive of failed vein puncture.

Background and Objectives It is generally accepted that the malignancy of pancreatic cancer is dependent upon the extent of invasion as well as metastasis. However, the factors and mechanisms are incompletely understood. We investigated whether glial cell lined-derived neurotrophic factor (GDNF) enhances the invasive and adhesive behaviors of pancreatic cancer cells by altering of the expression of integrins. Methods The expression of the GDNF receptor in pancreatic cancer cell lines (SW1990 and Capan-2) was confirmed by RT-PCR. Then we determined the expression of integrin subunits and the alteration of their expression by GDNF using flow-cytometric analysis and a cellular enzyme-linked immunosorbent assay (CELISA). Adhesion and invasion assay were performed to investigate whether increased integrin expression affected the interaction between cancer cells and ECM proteins. Results The GDNF receptor subunits were expressed in pancreatic cancer cells. GDNF enhanced the expression of some of the integrin subunits and increased their adhesive and invasive abilities. The enhanced expression and associated increase in adhesive and invasive abilities were inhibited by blocking the GDNF receptor or the integrin β1 subunit. Conclusion The enhancement of integrin expression by GDNF signaling through the GDNF receptor strongly influences invasion and adhesion to ECM proteins by pancreatic cancer cells. J. Surg. Oncol. 2005;91:77–83. © 2005 Wiley-Liss, Inc.

OBJECTIVES The invasive potential is increased by glial cell line-derived neurotrophic factor (GDNF) in human pancreatic cancer cell lines. We researched whether the signaling pathway activated by GDNF correlates with the nuclear factor-kappaB (NF-kappaB) in human pancreatic cancer cell lines and whether the inhibition of NF-kappaB activity is associated with suppression of invasive potential. METHODS Proliferation of human pancreatic cancer cell lines (BxPC-3 and MIA PaCa-2) was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays (MTT assay). NF-kappaB activity was examined by dual luciferase assay and electrophoretic mobility shift assay. In addition, to investigate the invasive potential, an in vitro invasion assay was performed. RESULTS Proliferation of both cell lines was decreased by a proteasome inhibitor, MG132, in a dose-dependent manner, but proliferation of control and IkappaBalphaM vector-transfected BxPC-3 cells was similar. The invasion cell number and the NF-kappaB activity were increased by GDNF stimulation. However, in the presence of MG132 or IkappaBalphaM, which blocks the nuclear localization of NF-kappaB, both were significantly suppressed. Furthermore, reduced activity of both remained unchanged by GDNF stimulation. CONCLUSION These results indicate that GDNF promotes NF-kappaB activation and that the latter is involved in the invasive potential of human pancreatic cancer cells.

To investigate the mechanisms of metastasis formation in human pancreatic carcinoma, we examined the angiogenic capabilities of human pancreatic cancer cell lines with different metastatic potentials and the roles of inflammatory cytokines.Interleukin (IL)-8 secretion by human pancreatic cancer cells stimulated with IL-1alpha or IL-1 receptor antagonist (IL-1ra) was measured by enzyme-linked immunosorbent assay (ELISA). We then examined how cancer cells with different metastatic potentials influenced the proliferation and tube formation of human umbilical vein endothelial cells (HUVECs) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye reduction method (MTT assay) and an angiogenesis assay, respectively. We also examined the role of inflammatory cytokines in relation to tumor metastatic potential and angiogenesis.IL-8 secretion levels by pancreatic cancer cells were regulated by IL-1alpha and correlated with metastatic potential. Both HUVEC proliferation and tube formation were strongly enhanced by coculture with metastatic pancreatic cancer cells and were enhanced to a similar extent by culture in the presence of IL-1alpha and IL-8. In contrast, blockade of IL-1alpha or IL-8 inhibited HUVEC proliferation and angiogenesis.The inflammatory cytokines IL-1alpha and IL-8 may have an important role in metastasis via vascular endothelial cell proliferation and angiogenesis.

To investigate the malignancy and differentiation of intraductal papillary-mucinous tumors (IPMTs) and mucinous cystic tumors (MCTs) of the pancreas, clinicopathologic characteristics and immunohistochemical features were analyzed.The clinicopathologic characteristics and immunohistochemical features of 24 patients with IPMT and 8 with MCT who underwent pancreatic resections at our hospital were examined. Immunohistochemical features analyzed included expression of p53 protein, proliferating cell nuclear antigen, integrins, interleukin-1 receptor type I, and hormone-associated receptors, and the factors correlated with malignancy were identified by multiple logistic regression.Among the IPMTs, there were 16 intraductal papillary adenomas, 5 intraductal papillary adenocarcinomas, and 3 moderate dysplasias. Among the MCTs, there were 6 mucinous cyst adenomas and 2 mucinous cyst adenocarcinomas. Multivariate analysis revealed that of the clinicopathologic characteristics, only the presence of mural nodules (odds ratio (OR) 7.12, P = 0.044) was independently correlated with the malignancy of IPMTs, and that of the immunohistochemical features, only alpha integrin subunit expression was independently correlated with malignancy of pancreatic mucinous tumors (OR 15.6, P = 0.036), especially IPMTs (OR 35.7, P = 0.012).These results indicate that alpha-containing integrin expression can be a significant marker of malignancy in pancreatic mucinous tumors.

INTRODUCTION AND AIMS Intraductal papillary-mucinous tumors (IPMTs) of the pancreas are intraductal tumors with diffuse or segmental dilation of the pancreatic ducts and intraductal papillary growth with abundant mucous secretion. We examined clinicopathological features and immunohistochemical findings to investigate the malignancy of IPMTs. METHODOLOGY Between April 1994 and December 2002, 23 patients with IPMT underwent pancreatic resections at Nagoya City University Hospital, Japan. We immunohistochemically analyzed the expression of p53 protein, proliferating cell nuclear antigen, alpha6-integrin subunit, alpha5beta1 integrin, and interleukin-1 receptor type I in tumor specimens from the 23 patients with IPMT. RESULTS The tumors were classified as intraductal papillary adenoma (n = 16), intraductal papillary adenocarcinoma and moderate dysplasia (n = 7). At a median follow-up of 42.9 months, 2 patients had died of this disease. The actuarial 5-year disease-free survival rate was 80.7%. Expression of the alpha6-integrin subunit was significantly strong in adenocarcinoma and moderate dysplasia tissues of IPMTs (P = 0.038). CONCLUSIONS Our current results indicate that alpha6-containing integrin expression can be a significant marker of malignancy in IPMTs. We emphasize that immunohistochemical investigation of resected specimens is indispensable in cases of IPMT, so that appropriate postoperative treatments for malignant IPMTs are initiated to improve prognosis.

Introduction Pancreatic cancer cells express a number of functionally active integrins that are related to their adhesive and invasive abilities. Aims To determine whether glial cell line-derived neurotrophic factor (GDNF) influences the expression of integrins in pancreatic cancer cell lines and to elucidate the mechanisms of adhesion and invasion to extracellular matrix (ECM) proteins. Methodology The expression of integrin subunits and the alteration of their expression by GDNF were examined by flow-cytometric analysis and cellular enzyme-linked immunosorbent assay in pancreatic cancer cell lines (MIA PaCa-2 and BxPC-3). Assays of adhesion and invasion of cancer cells to ECM proteins were conducted to investigate whether increased integrin expression affects the interaction between cancer cells and putative integrin ECM ligands. Results Expression of some of the integrin subunits in pancreatic cancer cells was enhanced by GDNF. The enhancement and associated increase in adhesive and invasive ability by GDNF were inhibited by blocking the GDNF receptor or the integrin β1 subunit. Conclusions In pancreatic cancer, the enhancement of integrin expression by GDNF signaling through the GDNF receptor strongly influences adhesion and invasion to ECM proteins.

To Investigate the prognostic indicator, we examined the expression of α6- and α5- integrin and interleukin-1 receptor type I (IL-1RI) immunohistochemically, and analyzed the correlation between immunohistochemical findings and clinicopathological factors in pancreatic cancer. In patients with a strongly expressing α6- integrin subunit or weakly expressing α5β1-integrin in pancreatic cancer tissues there was a significant association with advanced TNM stage (P = 0.027 and 0.014, respectively), presence of liver metastases (P = 0.032 and 0.002, respectively), and poor prognosis (P = 0.0155 and 0.0056, respectively). In patients with a weakly expressing α6 integrin subunit or weakly expressing α5β1-integrin in noncancerous pancreatic tissues there was a significant association with poor prognosis (P = 0.0324 and 0.0396, respectively). Multivariate analysis demonstrated that strong expression of α6- and weak expression of α5β1-integrin were found to be independent prognosticators in pancreatic cancer patients. Our present results indicate that α6β1- and α5β1-integrin expression can be a significant prognostic indicator in pancreatic cancer.

Objective: To investigate the mechanisms of metastasis formation in metastatic human pancreatic cancer, we examined the enhancement in integrin expression, and adherence to and invasiveness into extracellular matrix (ECM) proteins of human pancreatic cancer cells after exposure to interleukin (IL)-1α. Methods: Expression of IL-1 receptor type I (IL-1RI) and alterations in integrin subunits by IL-1α were examined by flow-cytometric analysis and by cellular enzyme-linked immunosorbent assay in four human pancreatic cancer cell lines (BxPC-3, PaCa-2, PANC-1, and SW1990), respectively. In addition, assays of cancer cell adhesion and invasion to ECM proteins were performed to investigate whether increased integrin expression affected the adhesive and invasive interaction between cancer cells and the putative integrin ECM ligands. Furthermore, immunohistochemistry was used to assess integrins and IL-1R1 expression in pancreatic tissues. Results: In metastatic cancer cells, expression of the α6 subunit was enhanced by IL-1α treatment. While metastatic cancer cells exhibited preferential adherence to and invasion into laminin, these properties were enhanced by IL-1α. The α6 subunit and IL-1RI were strongly expressed in pancreatic tissues from pancreatic cancer patients with liver metastasis. Conclusions: In pancreatic cancer, IL-1α enhanced α6β1-integrin expression, probably via increased IL-1RI levels. Our results indicated that α6β1-integrin and IL-1RI expression may play important roles in metastasis formation.

Background and Objectives Adhesion and invasion of tumor cells to extracellular matrix (ECM) proteins play an important role in tumor metastasis formation. We investigated the enhancement of adhesive and invasive behavior to ECM proteins of human pancreatic cancer cells by interleukin-1α (IL-1α) to examine the mechanism of adhesion and invasion of metastatic human pancreatic cancer cells to ECM proteins. Methods The enhancement of integrin subunits by IL-1α was examined by cellular enzyme-linked immunosorbent assay (CELISA) in two metastatic human pancreatic cancer cell lines (BxPC-3 and SW1990) and two nonmetastatic pancreatic cancer cell lines (PaCa-2 and PANC-1). In addition, assays of cancer cell adhesion and invasion to ECM proteins were performed to investigate whether increased integrin expression affected the invasive interaction between cancer cells and the putative integrin ECM ligands. Results Expression of the α6 subunit by metastatic cancer cells was enhanced by IL-1α. Metastatic cancer cells also exhibited preferential adherence and invasion to laminin compared with nonmetastatic cancer cells, and this was enhanced by IL-1α. Conclusions The enhancement of α6β1-integrin by Il-1α acting through IL-1RI, as well as the expression of α6β1-integrin, plays an important role in metastasis formation in pancreatic cancer. J. Surg. Oncol. 2003;82:51–56. © 2002 Wiley-Liss, Inc.

Microwave rotational spectra for normal and N -deuterated species of δ-valerolactam (2-piperidinone) were observed in the frequency range from 8 to 40 GHz. The spectra of the ground vibrational states for two isotopic species and the excited vibrational states for normal species were assigned, and the rotational constants (MHz) in ground vibrational state were determined using the Watson A -reduced Hamiltonian: A = 4590.96(11), B = 2495.03(2), C = 1731.06(2) for normal species and A = 4436.79(13), B = 2484.49(1), C = 1703.83(1) for deuterated species, respectively. The comparison of the rotational constants and r s coordinates of the imino hydrogen atom with ones from the ab initio MO calculation at the MP2/6-31G( d , p ) level of theory led to the conclusion that the spectra assigned were due to a half-chair conformer. One vibrationally excited state was observed and its vibrational frequency was 95(64) cm −1 .

バイオマスとして工業生産に利用されているChlorella sp.を, 従属栄養条件下で培養した.培養中のpHは細胞が対数増殖を維持できる培養初期は6.0, それ以後を8.0にシフトするパターンがクロロフィル (CHL) 生産には最適であることが判明した.しかしながら, 撹拌速度600r.p.m.では, この最適pHシフトにおいても最大CHL含量は1.28W/W%でしかなかった.他方, 撹拌は300r.p.m.以下の低速度の方が最大CHL含量の増加には有効で, 撹拌速度250r.p.m.において3.10W/W%と600r.p.m.の場合の約2.4倍の最大CHL含量に到達した.この影響は, 培養中期以後に酸素供給不足の期間が存在することに依存していることが示唆された.さらに, 撹拌速度を250r.p.m.とした場合, pHを6.0で一定に制御しても最大CHL含量は約3W/W%に到達したが, 培養中期のCHL比生産速度は0.54×10-3kg/ (kg dry cell・h) と最適pHシフトの場合の約70%であった.この結果より, pHシフトと撹拌速度の両者を加味することにより効率的なCHL生合成が可能になることが明らかになった.

Chlorella regularis strain YA-603 was previously found to exhibit a comparatively high growth rate and improved 5-aminolevulinic acid (ALA) productivity when cultured heterotrophically. Although the universal tetrapyrrole precursor ALA is synthesized from glutamate in algae and higher plants, in this study the addition of glutamate to the medium did not enhance ALA production in the heterotrophic culture of C. regularis YA-603. On the other hand, the addition of glycine, which is a precursor of ALA biosynthesis via the Shemin pathway in animal cells and some bacteria, enhanced both the specific growth rate and ALA production. Moreover, when a second glycine addition was made during the culture, the ALA concentration increased by about 1.5 times compared to that obtained with one glycine addition. From these results, it is suggested that the Shemin pathway contributes to ALA production, and that addition of glycine in the heterotrophic culture of C. regularis YA-603 can significantly increase the amount of ALA produced.

Effects of pH and temperature on 5-aminolevulinic acid(ALA) production were investigated in the heterotrophic cultures of newly screened Chlorella regularis YA-1. The optimum pH and temperature were 6.0 and 33°C, respectively. 1.96 mM of ALA was produced extracellularly by the YA-1 strain under the optimum conditions. The pH was found to be more critical than temperature for ALA production. Moreover, it was suggested that the activity of ALA production and the inhibitory effect of levulinic acid for ALA dehydratase were higher at a lower pH. One of the mutant strains, the YA-603 strain, obtained from the 1-methyl-3-nitro-1-nitoroso-guanidine (NTG) treatment of the YA-1 strain, demonstrated higher activity of ALA production than the YA-1 strain, and extracellularly produced 2.40 mM of ALA under the optimum conditions for the YA-1 Strain.

The effects of the various operating conditions were investigated on the production of β -carotene in the calutivation of carrot cells using an external loop airlift bubble column suitable to a larger scale production. The optimum of each operating variable was determined taking the content, yield and productivity of β - carotene as the objective functions. A simplified kinetic model was proposed to reproduce the observed time courses. The model assumes an inhibited cell growth of the Monod type and a nonlinear dependence of β -carotene production on growth. The kinetic parameters in the model were determined and correlated with the operating variables. The correlations were derived based on the maximum specific growth rate for the control culture, μ me representing an overall nature of the cell lines used. The kinetic model with the correlations obtained fairly reproduced the observed time courses and predicted the optimal cell lines as well as the optimal operating conditions.

Background and Objectives Interleukin (IL)-1α and hepatocyte growth factor (HGF) play an important role in pancreatic cancer proliferation, angiogenesis, and invasiveness. The aim of this study was to investigate the cooperative role of HGF and IL-1α in metastatic processes promoted by interactions between pancreatic cancer cells and stromal cells. Methods Expression of IL-1α and HGF mRNA and protein was determined by RT-PCR and ELISA. The effect of HGF on metastatic potential was evaluated by proliferation, invasion, and angiogenesis assays using an in vitro system consisting of co-cultured tumor cells and stromal cells. Results IL-1α expression was closely correlated with metastatic potential, and cancer cell-derived IL-1α significantly promoted HGF expression by fibroblasts (P

Purpose Spigelian hernia is a ventral abdominal hernia that only rarely causes incarceration or strangulation of the bowel. Few reports of laparoscopic treatment for Spigelian hernia have been published. Here we report a case of laparoscopic Spigelian and inguinal hernia repair using the Kugel Patch—a shape memory prosthesis. Methods The patient experienced severe intermittent lower abdominal pain, which was difficult to diagnose until repeated computed tomography scans were performed. The patient also had inguinal swelling, which had not been confirmed preoperatively; therefore, we repaired an inguinal hernia and a Spigelian hernia simultaneously by laparoscopic operation using the Kugel Patch. Results The surgery was completed without bleeding complications or converting to open operation. The patient was discharged from the hospital on postoperative day 4 with no complications. Conclusion An accurate preoperative diagnosis of Spigelian hernia led to the successful laparoscopic repair with the Kugel Patch.

Since angiogenesis enables solid tumors, including pancreatic cancer (PaCa), to grow and metastasize, the development of anti-angiogenic agents is currently one of the urgent issues. Proteasome inhibitors are well known for inhibiting nuclear factor-kappa B (NF-kappaB) activity in various cancer cells, but little is known about their biologic mechanisms against angiogenesis in PaCa. We divided human PaCa cell lines into high-angiogenic (BxPC-3 and SW 1990) and low-angiogenic (MIA PaCa-2 and Capan-2) groups. The high-angiogenic PaCa cell lines constitutively expressed high NF-kappaB activity and produced high levels of vascular endothelial growth factor (VEGF) and interleukin 8 (IL-8). The conditioned media from BxPC-3 significantly enhanced both proliferation of and tube formation by human umbilical vein endothelial cells (HUVECs) and these enhancements were significantly inhibited by the proteasome inhibitor MG132 treatment. Collectively, MG132 blocked PaCa-derived VEGF and IL-8 production through inhibition of NF-kappaB activity. Thus, proteasome inhibitors may prove beneficial as anti-angiogenic therapy for PaCa. Our studies show that MG132, a proteasome inhibitor, significantly blocked pancreatic-cancer-associated angiogenesis through inhibition of NF-kappaB and NF-kappaB-dependent proangiogenic gene products VEGF and IL-8.

Objectives: There is strong evidence for an important role of cyclooxygenase (COX) 2 and COX-2-generated PGE2 during pancreatic tumorigenesis. Cyclooxygenase 2 has therefore become a potential chemotherapeutic target for pancreatic cancer. However, recent studies raised concerns regarding the safety of selective COX-2 inhibitors. Although the benefits of COX-2 inhibition may eventually outweigh the associated cardiovascular risks, there are a number of alternative targets for inhibiting the formation of PGE2 in human tumors that may prove less harmful to the patient. This study aimed at analyzing the expression of various proteins involved in the generation of PGE2 in human pancreatic cancers. Methods and Results: Real-time polymerase chain reaction and Western blot analyses demonstrated overexpression of cytoplasmic phospholipase A2, COX-2, cytoplasmic prostaglandin E synthase, and microsomal prostaglandin E synthases 1 and 2 in most human pancreatic cancers when compared with matched normal pancreas. Immunohistochemistry revealed expression of these proteins predominantly by pancreatic cancer cells. Variable expression of these proteins was also confirmed in several human pancreatic cancer cell lines. Conclusions: Our studies demonstrated for the first time that various proteins involved in the generation of PGE2 are overexpressed in human pancreatic cancers. These proteins may represent potentially novel targets for the therapy of pancreatic cancers.

Type-1 insulin-like growth factor (IGF-1) up-regulates cell proliferation and invasiveness through activation of PI3K/Akt signaling pathway. IGF-1 also down-regulates the tumor suppressor chromosome 10 (PTEN). We investigated the mechanism by which IGF-1 affects cell proliferation and invasion by suppression of PTEN phosphorylation and interaction with PI3K/PTEN/Akt/NF-small ka, CyrillicB signaling pathway in pancreatic cancer.The expression of IGF-1 receptor (IGF-1R) and PTEN in five pancreatic cancer cell lines was determined by RT-PCR and Western blot. Proliferation and invasion were investigated by WST-1 assay and Matrigel-double chamber assay. Pancreatic cancer cells were transfected with PTEN siRNA to investigate which signaling pathway correlates in regulation of cancer cell proliferation and invasion.Five pancreatic cancer cell lines expressed PTEN and IGF-1R in mRNA and protein levels. Suppression of PTEN phosphorylation strongly enhanced cell proliferation and invasion stimulated with IGF-1 via activation of PI3K/Akt/NF-small ka, CyrillicB signaling pathway. In addition, knockdown of PTEN by siRNA transfection also enhanced activation of PI3K/Akt/NF-small ka, CyrillicB pathway, subsequently up-regulating cell invasiveness and proliferation.The IGF-1/PI3K/PTEN/Akt/NF-small ka, CyrillicB cascade may be a key pathway stimulating metastasis of pancreatic cancer cells. We suggest that interfering with the functions of IGF-1/PI3K/Akt/NF-small ka, CyrillicB might be a novel therapeutic approach to inhibit aggressive spread of pancreatic cancer.

Gastrointestinal stromal tumors (GISTs) are characterized by remarkable variability in their differentiation potential, but most of these lesions do not display convincing smooth muscle or neuronal differentiation. Here we report the case of a 65-year-old woman who underwent a perfect resection of a large submucosal tumor that displayed extragastric growth. The specimen was revealed to be an elastic soft tumor, 18 x 25 x 11 cm in size. Histologically, the tumor consisted of spindle-shaped cells, with a mitotic rate of 12 per 10 high-power fields. Immunohistochemically, the tumor showed positive staining for CD34 and c-kit but negative staining for alpha-smooth muscle actin, Desmin, and s-100 protein. From these findings, the tumor was diagnosed as an uncommitted type of GIST with high-grade malignancy. This case needs careful and long-term follow-up to monitor for signs of local recurrence or distant metastasis.

Background The tumour suppressor phosphatase and tensin homolog (PTEN) is an important negative regulator of cell-survival signaling. To evaluate the correlation between PTEN expression and clinicopathological characteristics of colorectal cancer patients with and without liver metastases, we investigated PTEN expression in primary colorectal cancer and colorectal cancer liver metastases. Methods Sixty-nine pairs of primary colorectal cancer and corresponding liver metastasis specimens were analyzed immunohistochemically, and the correlation between immunohistochemical findings and clinicopathological factors was investigated. Seventy primary colorectal cancer specimens from patients without liver metastases were used as controls. Results PTEN was strongly expressed in 44 (62.9%) colorectal cancer specimens from patients without liver metastases. In contrast, PTEN was weakly expressed in 52 (75.4%) primary colorectal cancer specimens from patients with liver metastases, and was absent in liver metastases. Weak PTEN expression in colorectal cancer tissues was significantly associated with advanced TNM stage (p < 0.01) and lymph node metastasis (p < 0.05). PTEN expression was significantly stronger in primary colorectal cancer specimens from patients without liver metastases. Furthermore, among colorectal cancer patients with liver metastases, the 5-year survival rate was significantly higher in patients with positive PTEN expression compared to those with negative PTEN expression (p = 0.012). Conclusion Our results suggest that loss of PTEN expression is involved with colorectal cancer aggressive capacity and that diagnostic evaluation of PTEN expression may provide valuable prognostic information to aid treatment strategies for colorectal cancer patients.

Background Granulocyte-colony stimulating factor (G-CSF)-producing cancer has been reported to occur in various organs, especially the lung. However, G-CSF-producing colorectal cancer (CRC) has never been reported in the English literature. Case presentation A 57-year-old man was admitted for the surgical removal of a rectal cancer. Some hepatic tumors in the liver were revealed concurrently, and their appearance suggested multiple liver metastases. Low anterior resection was performed. with the help of histopathological examination and immunohistochemical studies, we diagnosed this case to be an undifferentiated carcinoma of the rectum. After the operation, the white blood cell (WBC) count increased gradually to 81,000 cells/μL. Modified-FOLFOX6 therapy was initiated to treat the liver metastases, but there was no effect, and peritoneal dissemination had also occurred. The serum level of G-CSF was elevated to 840 pg/mL (normal range, Conclusion This is the first case of G-CSF-producing rectal cancer, and its prognosis was very poor.

Previously we reported the critical role of interleukin (IL)-1alpha in liver metastasis from pancreatic cancer (PaCa). However, its role in angiogenesis and metastasis, particularly as it relates to the interaction between tumor and stromal cells, was not clearly elucidated. To further investigate, we initially compared vascular endothelial cell migration and tube formation in human PaCa cell lines that differed in metastatic potential. We then compared the effects of IL-1alpha derived from PaCa cells on the same processes.Expression of IL-1alpha mRNA and protein in PaCa cells was measured by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. IL-1alpha mRNA and protein were detected only in predominantly liver-metastatic PaCa cells. Next, we examined differences in migration and tube formation by human umbilical vein endothelial cells (HUVECs) in the various PaCa cells using invasion and angiogenesis assays, respectively. Furthermore, we determined the effects of IL-1alpha secreted by PaCa cells on migration and tube formation by HUVECs using coculture experiments.Expression of IL-1alpha mRNA and protein was observed only by the highly liver-metastatic PaCa cell lines BxPC-3 and SW 1990. Both HUVEC migration and tube formation were significantly enhanced by coculture with metastatic PaCa cells and IL-1alpha (P0.01). Similarly, blockade of IL-1alpha by its antagonist inhibited HUVEC migration and tube formation (P0.01).Our results indicate that IL-1alpha secreted by PaCa cells plays an important role in metastasis through vascular endothelial cell invasion and angiogenesis. Thus, blocking IL-1alpha is a potential novel therapeutic strategy in PaCa.

The production of xanthan gum by Xanthomonas campestris ATCC 13951 in a fermentor using twin turbine-type impellers with 6 flat blades was studied to determine the effects of impeller spacing on the specific production rate, the volume of the high shear stress region around the impellers, and the agitation power requirements, and a comparison was made with the use of a single impeller. The following results were obtained. (i) The specific rate of xanthan gum production was dependant on the volume of the high shear region in the fermentor, even in the twin-impeller system. (ii) The greater was the space between the two impellers, the larger was the volume of the high shear region. (iii) The volume of the high shear region and the specific rate of xanthan gum production were greater with two impellers than with a single impeller, even if the agitation rate was adjusted to consume the same power. These results strongly indicate that the production of xanthan gum depends on the overall volume of the high shear region, which is enhanced by duplication of impellers, and that a twin-impeller system is also economically advantageous in terms of the power consumption required for agitation.

This study was performed to determine whether GDNF influences the expression of integrins in colorectal cancer cell lines and to elucidate the mechanisms of adhesion to and invasion of extracellular matrix (ECM) proteins.The expression of integrin subunits and alteration of this expression by GDNF were examined by flow-cytometric analysis and cellular enzyme-linked immunosorbent assay in four human colorectal cancer cell lines. Assays to evaluate adhesion and invasion of cancer cells toward ECM proteins were conducted to investigate whether increased integrin expression affects the interaction between cancer cells and putative integrin ECM ligands.The RET/GFRalpha-1 receptor complex for GDNF was expressed in all four colorectal cancer cell lines. The expression of the Beta1 integrin subunit in these cells was significantly enhanced by GDNF. The enhancement and associated increase in adhesion and invasion abilities in response to by GDNF were inhibited by blocking the GDNF receptor or the integrin P1 subunit.In colorectal cancer, the enhancement of integrin expression by signaling through the GDNF receptor strongly influences adhesion to and invasion of ECM proteins.

The process of tumor progression and metastasis involves degradation of the extracellular matrix and is governed by an intricate balance of proteases, their activators and their inhibitors, in which malignant cells are permitted to infiltrate the adjacent structures and gain access to lymph and blood vessels. These proteases can be broadly categorized into three families: matrix metalloproteinases, serine proteinases and cysteine proteinases, all of which have all been implicated in these processes. The presence of neural invasion is often considered to be a poor prognostic sign; however, the cellular mechanisms underlying this propensity for perineural invasion are unknown. We recently researched the relationship between the glial cell line-derived neurotrophic factor and perineural invasion by human pancreatic cancer cells. We also confirmed that NF-kappa B is a part of the signaling pathway from the glial cell line-derived neurotrophic factor in human pancreatic cancer cells, and documented the inhibitory effect of gabexate mesilate, a well-known non-physiological synthetic serine protease inhibitor, for pancreatic cancer invasion. Recent studies on the role of proteases and protease inhibitors in pancreatic cancer invasion are also reviewed.

Pancreatic ductal adenocarcinomas are thought to arise from noninvasive, intraductal precursor lesions called pancreatic intraepithelial neoplasias (PanIN). The study of PanINs holds great promise for the identification of early detection markers and effective cancer-preventing strategies. Cyclooxygenase-2 (COX-2) represents an intriguing target for therapeutic and preventive approaches in various human malignancies. The aim of the present study was to evaluate the efficacy of a selective COX-2 inhibitor to prevent the progression of PanINs in a conditional KrasG12D mouse model. Offspring of LSL-KRASG12D x PDX-1-Cre intercrosses were randomly allocated to a diet supplemented with the selective COX-2 inhibitor nimesulide (400 ppm) or a control diet. After 10 months, animals were sacrificed. Successful recombination in the pancreas was evaluated by PCR. The pancreas of KRASG12D;PDX-1-Cre mice was analyzed for the presence of murine PanINs. Animals fed the COX-2 inhibitor had significantly fewer PanIN-2 and PanIN-3 lesions than control animals (P < 0.05). Ten percent of all pancreatic ducts in the nimesulide-fed animals showed PanIN-2 or PanIN-3 lesions, whereas 40% of the pancreatic ducts in the control animals had PanIN-2 or PanIN-3 lesions. Intrapancreatic prostaglandin E2 levels were reduced in nimesulide-fed animals. Immunohistochemistry confirmed COX-2 expression in early and late PanINs. In summary, we found that the selective COX-2 inhibitor nimesulide delays the progression of pancreatic cancer precursor lesions in a preclinical animal model. These data highlight the importance of COX-2 in the development of pancreatic cancer. Inhibition of COX-2 may represent an intriguing strategy to prevent pancreatic cancer in high-risk patients. [Cancer Res 2007;67(15):7068–71]

Gastrointestinal stromal tumors (GISTs) have unique immunohistochemical and molecular genetic features that set them apart from leiomyomas, leiomyosarcomas, and schwannomas. Although recurrence of GIST usually tends to develop locally or in the liver, rectal GIST reoccur predominantly at the original site of the tumor. We describe a rare case of rectal GIST with multiple liver metastases. We carried out immunohistochemical staining for p53 protein, proliferating cell nuclear antigen (PCNA), integrins, and interleukin-1 receptor type I (IL-1RI) in order to investigate the degree of malignancy of this neoplasm in addition to the immunohistochemical analyses that were necessary for diagnosing GIST. Histologically, the rectal tumor was classified as an uncommitted type of rectal GIST with multiple liver metastases. Positive immunostaining for PCNA, alpha6 integrin subunit, and IL-1RI was found in both the rectal and hepatic tumors. The patients with a rectal GIST may have an increased risk of liver metastasis and a poor prognosis independent of the size of the tumor.

Background To better understand the underlying mechanism of liver metastasis formation in human gastric cancer, we evaluated the angiogenic capabilities of human gastric cancer cell lines with different metastatic potentials as well as the role of interleukin (IL)-1α in the angiogenic process. Materials and methods Reverse transcription-polymerase chain reaction was used to detect the expression of IL-1α and vascular endothelial growth factor (VEGF) mRNA in gastric cancer cell lines with different liver metastatic potentials. Levels of VEGF secreted by human gastric cancer cells were measured by enzyme-linked immunosorbent assay. We also examined how gastric cancer cells with different metastatic potentials influence the proliferation and tube formation of human umbilical vein endothelial cells (HUVECs) using the Premix WST-1 cell proliferation assay system and an angiogenesis assay, respectively. Results IL-1α expression levels were significantly correlated with liver metastatic potential in gastric cancer cell lines. Levels of VEGF secreted by gastric cancer cells appear to be regulated by IL-1α through IL-1 receptor Type 1 and were correlated with liver metastatic potential. Both HUVEC proliferation and tube formation were strongly enhanced by coculture with high liver-metastatic gastric cancer cells and were enhanced to a similar extent by culture in the presence of IL-1α. In contrast, blockade of IL-1α inhibited both HUVEC proliferation and angiogenesis. Conclusions IL-1α may play a role in liver metastasis of gastric cancer via enhanced vascular endothelial cell proliferation and angiogenesis.

Background The invasive interaction between cells and their matrix has important roles in tumor cell invasion. This study investigated modulation of basement membrane (BM) proteins, especially collagen IV (Coll IV), laminin, and fibronectin (FN), in invasion of human pancreatic cancer cells. Furthermore, we examined the roles of β 1 -integrins and arginine-glycine-aspartic (RGD)-containing oligopeptide in cell–matrix interactions. Materials and methods Expression of integrins were examined by reverse transcriptase-polymerase chain reaction and flow-cytometric analysis in three human pancreatic cancer cell lines (BxPC-3, PANC-1, and SW1990), respectively. To determine the effect of BM proteins, invasion assays were performed. Western blot analysis for extracellular signal-regulated kinase (ERK) was performed to investigate the involvement of ERK1/2 signaling pathways. Results BM proteins significantly enhanced the invasive behavior of pancreatic cancer cells. Pretreatment with anti-β 1 -integrin antibody suppressed invasion into Matrigel, but RGD-containing peptide inhibited invasion, which was enhanced by Coll IV and FN, not laminin. Treatment with both RGD-containing peptide and β 1 -integrin antibody inhibited ERK1/2 phosphorylation activated by Coll IV and FN. Conclusions BM proteins have positive actions on the processes of pancreatic cancer cell invasion and cross-talk between BM proteins and β 1 -integrins widely participates in the multistep processes of pancreatic cancer invasion and metastasis formation.

We analyzed clinicopathologic and imaging findings of intraductal papillary-mucinous tumors (IPMTs) and mucinous cystic tumors (MCTs) of the pancreas to evaluate the difference between IPMTs and MCTs, and to identify the signs indicative of malignancy in IPMTs.Clinicopathological features of 20 patients with IPMT and six patients with MCT of the pancreas were studied.The patients with IPMT comprised 16 males and four females with a mean age of 62.9 years. Eighty percent of IPMTs were located in the pancreatic head, and the mean tumor size was 38.6mm. Recurrence was observed in one patient, who died of IPM adenocarcinoma. In contrast, all patients with MCT were females, with a mean age of 53.0 years. None of the MCTs arose in the pancreatic head, and the mean tumor size was 42.7mm. One patient died of MC adenocarcinoma, but all of the others survived without recurrence. The difference in gender, location of the tumor, and connection to the pancreatic duct reached statistical significance between IPMTs and MCTs. A significant connection to the pancreatic duct and high level of serum carbohydrate antigen 19-9 (CA19-9) was observed in the adenocarcinoma and moderate dysplasia groups of IPMT.The main duct type and an elevation of serum CA19-9 level suggested malignancy in IPMTs.

The high incidence of anastomotic stenosis after gastrointestinal surgery using circular staplers is a major problem. In response, we have developed a new technique that uses a linear stapler to enlarge an anastomotic opening made using a circular stapler. Anastomoses were created by the new technique or by the conventional approach using a circular stapler in pig small intestine. The method was also applied in treatment of a colon cancer patient. The area of the anastomotic opening obtained with the new technique was more than 3 times that in the control (p