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We studied potential indicators of severe acute pancreatitis by measuring the blood concentrations of various cytokines, polymorphonuclear leucocyte elastase (PMN-E), acute phase reactants, pancreatic amylase (P-AMY), pancreatic elastase-1 (E-1) and white blood cell (WBC) counts in patients with acute pancreatitis. In addition, the presence of multiple organ damage was assessed. Subjects consisted of 22 patients with acute pancreatitis including severe (n = 11), moderate (n = 4) and mild (n = 7) cases. A significant positive correlation was observed between the number of organs damaged and the peak concentrations of interleukin (IL)-6, PMN-E, C-reactive protein (CRP) and pancreatic secretory trypsin inhibitor (PSTI). Among these markers, blood concentrations of PMN-E and IL-6 rapidly increased and peaked at the early phase of acute pancreatitis whereas CRP and PSTI did not. The elevation of PMN-E and IL-6 was greater the more severe the symptoms. However, no significant correlation was observed between the number of organs damaged and the maximum serum concentrations of P-AMY and E-1, or the WBC count, which have been considered to be markers of pancreatitis. These results suggest that PMN-E and IL-6 concentrations are useful indicators of severity and prognosis and their determination facilitates the selection of appropriate treatment in the early stages of disease to prevent the aggressive progression of acute pancreatitis. [ABSTRACT FROM AUTHOR]

We conducted clinical trials in patients with acute lung injury (ALI) associated with systemic inflammatory response syndrome using a selective neutrophil elastase inhibitor, sivelestat sodium hydrate (Sivelestat), to investigate the involvement of neutrophil elastase in ALI. In the phase III double-blind study (Study 1) in 230 patients, the efficacy of Sivelestat was evaluated with the pulmonary function improvement (PFI) rating as the primary endpoint, and the weaning rate from mechanical ventilator, the discharge rate from intensive care unit (ICU), and the survival rate as secondary endpoints. Afterwards, an unblinded study (Study 2) in 20 patients was conducted using procedures for weaning from mechanical ventilation to reevaluate its efficacy with ventilator-free days (VFD) value, the primary endpoint, and to compare with that of Study 1 subgroup, which met the selection criteria used in Study 2. Sivelestat increased PFI rating, reduced duration of mechanical ventilation, and shortened stay in ICU in Study 1, although there was no significant efficacy on the survival rate. VFD value in Study 2 was comparable to that in the optimal-dose group of Study 1 subgroup, and increase in VFD value correlated with PFI rating and increase in ICU free days. It was concluded that neutrophil elastase may be involved in the pathogenesis of ALI in humans. [Copyright &y& Elsevier]

The case of a 95-year-old Japanese female with a giant intramural tumor located in the upper third of the stomach is reported. A preoperative study using an endoscopic examination and gastrointestinal series revealed a large submucosal tumor with a deep concavity in the center of the surface accompanied by continuous bleeding. The tumor was removed by a minimal laparotomy instead of laparoscopic surgery because of its large size (8 × 7 × 6 cm). Its tissues were sampled, and examined by light microscopy, immunohistochemistry and electron microscopy. The tumor was vaguely encapsulated but had foci of partial infiltration of the capsule. It was comprised of spindle cells with severe nuclear pleomorphism. The mitotic count was four per 50 high-power fields. The tumor was stained strongly positive for vimentin, CD 34 and c-Kit, and weakly positive for neuron-specific enolase; had fine granular cytoplasmic stainings for synaptophysin; and was negative for cytokeratins, smooth muscle actin and S-100 protein. These immunohistochemical results strongly suggest that this tumor was a gastrointestinal autonomic nerve tumor (GANT); the ultrastructural findings, however, had no electron microscopic characteristics of the GANT. [ABSTRACT FROM AUTHOR]

Discusses the single nucleotide polymorphisms (SNP) and disease-susceptibility genes in digestive organ diseases. Use of SNP as DNA markers for population genetics; Genotypic risks associated with variants of SNP.

A letter to the editor is presented in response to an article about increased plasma concentrations of endothelin-1 in acute myocardial infarction.

Abstract: In squid jigging operations for neon flying squid (Ommastrephes bartramii), 30–40% of hooked squid are estimated to fall off (i.e., become detached) from the jigs during the drum-hauling process, caused by breakage/severance of the hooked tentacle(s). Reducing fall-off events can lead to higher productivity and more efficient use of the squid resource. In the present study, there were 950 fall-off events either directly observed or assessed from the residue of tentacle(s) left on the jigs compared with the 1720 total captures, which comprises 35.6% of 2670 total hooked squids. The fall-off ratio according to the mantle length (ML) was examined using logistic curve analysis and a higher fall-off ratio for smaller squid was confirmed through size selectivity curve analysis. F 50, the 50% probability of fall off from the jig, was estimated to be 37.4cm ML. The breaking strength of a single tentacle was determined to be similar to the body weight (BW) of squid smaller than 41.4cm ML, indicating a high possibility of tentacle breakage in the case of 1 tentacle grabbing the jig for smaller-sized squid. [Copyright &y& Elsevier]

In order to estimate exactly changes in the murine peristaltic movement by acetylcholine (ACh), we detected the circular muscle activity in the murine small intestine by measuring the changes in intraluminal pressure of the duodenal segment in vitro. When ACh was applied into the tissue baths, the basal tone of the duodenal preparation along the longitudinal axis increased, whereas the intraluminal pressure that reflected circular muscle activity decreased in a dose-dependent manner. Thus, the circular muscle layer of the murine duodenum relaxed by ACh. The circular relaxations by ACh were also observed in the other parts (jejunum and ileum) of the murine small intestine. The ACh-induced fall in the intraluminal pressure of the murine duodenal preparation was antagonized by atropine (10−6 M), not tetrodotoxin (10−6 M). The results indicate ACh acts as not only the excitatory transmitter for the longitudinal muscle but also the inhibitory transmitter for the circular muscle, and suggest that the functional receptors to ACh are different between longitudinal and circular muscle cells in the murine small intestine. [Copyright &y& Elsevier]

Introduction: The pancreatic secretory trypsin inhibitor (PSTI) is thought to be a specific inactivation factor of intrapancreatic trypsin activity. N34S mutation of PSTI was reported in chronic pancreatitis patients from all over the world including us, and is thought to be one of the predisposing factors for pancreatitis. The N34S mutation located close to the reactive lysine–isoleucine site (K41∼I42) might lead to a decreased inhibitory capacity. N34S is also in complete linkage with other intronic sequence variants such as IVS1-37T→C and IVS3-65insTTTT. Materials/Methods: Based on the hypothesis above, we performed biochemical experiments. (I) Activity of recombinant PSTI protein with N34S was compared to that of the wild type. (II) Molecular weight of PSTI from a patient with N34S was analysed by Western blotting to check the possibility of abnormal splicing (e.g. splice-out of exon 4). Results: (I) The function of N34S PSTI remained unchanged compared to wild-type PSTI. (II) The molecular weight of PSTI protein from a patient with N34S was not altered. Conclusion: We could not detect any functional or configurational abnormalities of N34S PSTI protein by the two experiments mentioned above. Other factors, such as translational abnormality by intronic mutation, may be associated with the predisposition to pancreatitis. [Copyright &y& Elsevier]

There are indications of the possible effects of sex hormones on pancreatic carcinoma. Estrogen receptor (ER) has been demonstrated in pancreatic tumors in experimental animals and in humans and it has been suggested that endocrine manipulation may be effective in the treatment of pancreatic carcinoma. However, it is still controversial whether this lethal cancer can potentially benefit from endocrine therapy. One explanation for the conflicting data on the benefit of hormonal manipulation in the treatment of pancreatic carcinoma may stem from the fact that there is no adequate marker to assess estrogen dependency of the pancreatic tumors. In this article, we review our work on tissue plasminogen activator as a prognostic guide to evaluate the efficacy of hormonal therapy in human pancreatic carcinoma, and also suggest that a selected subgroup of patients with this lethal cancer may have a potential clinical benefit from endocrine therapy, especially medroxyprogesterone acetate treatment. [ABSTRACT FROM AUTHOR]

Declines in landings of the hairtail Trichiurus japonicas indicate the need for more effective management of this species. Hairtail spawning peaks occur twice yearly in the Bungo Channel, in spring and autumn. Relationships between hairtail stock and brood seasonality were examined to determine if an association between either and a decline in landings existed. Stock assessments show that the biomass of both spring and autumn hairtail broods from within and around the Bungo Channel are decreasing, with a rapid reduction in spring-brood stock abundance after 2007 largely responsible for decreased landings. Yield and spawning per recruitment analyses indicate current fishing pressure to be higher than several reference points. We suggest that fishing pressure needs to be reduced by at least 20% of the current level for this fishery to remain sustainable, as the projected stock abundance and catch demonstrate that the current fishing pressure is unsustainable. Analysis of time-series data of recruits per spawning revealed spring-brood recruitment to have been strong in year classes 2003 and 2005. Of various options available for improved management of this fishery, we propose that fishing pressure should be reduced in the years following the appearance of strong year classes to increase future biomasses and landings. [ABSTRACT FROM AUTHOR]

Under the coastal fisheries co-management regime in Japan, local fishers play important roles in deciding on and implementing local management measures. Therefore, using the case study of a hairtail trolling line fishery, we conducted a transdisciplinary research with fishers, processors, government officers, etc. Taking social and biological factors into account, we defined management criteria and targets (resource, economic, human community, and local food culture) in collaboration with local fishers. We chartered local fishing vessels to determine parameters that were easily understood by local fishers. We established a local consultative committee comprising fishers, processors, and distributors, and developed a fish distribution strategy to nurture the local hairtail food culture. The integrated effects of various management scenarios were evaluated by the operating model. We found that co-ordinated management by both the trolling line fishery and purse seiners in the neighbouring prefecture was required to achieve targets. Two realistic scenarios are recommended: a conservative scenario that could achieve targets even with few strong year classes, and an adaptive scenario that protects strong year classes, but relies on a higher frequency of these. We shared these results with the local fishers and government officers, which led to actual improvements in management measures. [ABSTRACT FROM AUTHOR]

Overfishing has reduced the stock of hairtail Trichiurus japonicus around the Bungo Channel, Japan. To determine whether using larger bait in the trolling line fishery could avoid catching small/undersize hairtail, we developed and trialled a new large artificial bait (a soft plastic bait, 6 inches long). A traditional lure with natural bait (type-1), a new lure of the new artificial bait hook without natural bait (type-2), and a new lure with natural bait (type-3) were tested in fishing operations around the Bungo Channel. Compared with type-1, type-2 and type-3 caught fewer undersized and immature female individuals. The number of fish caught, yield, and composition of commercial size grades per recruitment were calculated from field data for each lure and compared. The catch sizes (number of fish per recruitment) were smaller for type-2 and type-3 than for type-1, but yield per recruitment was higher for type-3 than for type-1. Compared with type-1, type-2 and type-3 caught more large individuals, which are more valuable. The newly developed artificial bait conserves hairtail stocks by targeting larger fish, which is economically beneficial for the hairtail trolling line fishery. [ABSTRACT FROM AUTHOR]

We investigated the impact of curcumin on neutrophils. Chemotactic activity via human recombinant IL-8 (hrIL-8) was significantly inhibited by curcumin. Curcumin reduced calcium ion flow induced by internalization of the IL-8 receptor. We analyzed flow cytometry to evaluate the status of the IL-8 receptor after curcumin treatment. The change in the distribution of receptors intracellularly and on the cell surface suggested that curcumin may affect the receptor trafficking pathway intracellulary. Rab11 is a low molecular weight G protein associated with the CXCR recycling pathway. Following curcumin treatment, immunoprecipitation studies showed that the IL-8 receptor was associated with larger amounts of active Rab11 than that in control cells. These data suggest that curcumin induces the stacking of the Rab11 vesicle complex with CXCR1 and CXCR2 in the endocytic pathway. The mechanism for antiinflammatory response by curcumin may involve unique regulation of the Rab11 trafficking molecule in recycling of IL-8 receptors. [ABSTRACT FROM AUTHOR]

We investigated the impact of curcumin on neutrophils. Chemotactic activity via human recombinant IL-8 (hrIL-8) was significantly inhibited by curcumin. Curcumin reduced calcium ion flow induced by internalization of the IL-8 receptor. We analyzed flow cytometry to evaluate the status of the IL-8 receptor after curcumin treatment. The change in the distribution of receptors intracellularly and on the cell surface suggested that curcumin may affect the receptor trafficking pathway intracellulary. Rab11 is a low molecular weight G protein associated with the CXCR recycling pathway. Following curcumin treatment, immunoprecipitation studies showed that the IL-8 receptor was associated with larger amounts of active Rab11 than that in control cells. These data suggest that curcumin induces the stacking of the Rab11 vesicle complex with CXCR1 and CXCR2 in the endocytic pathway. The mechanism for antiinflammatory response by curcumin may involve unique regulation of the Rab11 trafficking molecule in recycling of IL-8 receptors. [ABSTRACT FROM AUTHOR]

Background: The pre-transplant administration of donor antigens to recipients is reported to prolong transplanted organ survival. We investigated the effect of pre-transplant intraportal administration of recipient blood on rat hepatic allograft survival. Materials and methods: Male LEW (RT1l) and ACI (RT1a) rats were used as transplant recipients and donors, respectively. Before transplantation, donors were transfused with recipient blood. Experimental animals were divided into groups as follows: group I, no treatment; group II, pre-treatment with recipient blood via the penile vein 7 days before transplantation; group III, pre-treatment with recipient blood via the portal vein 5 days before transplantation; and group IV, pre-treatment with recipient blood via the portal vein 7 days before transplantation. Serum interferon (IFN)-γ concentrations were measured post-operatively. Results: Animals in group I survived a mean of 10.1 ± 0.7 days. The survival of groups II and III was 10.6 ± 1.6 and 13.1 ± 0.9 days, respectively. The survival rate in group IV was prolonged significantly to 33.7 ± 2.6 days. Serum concentrations of IFN-γ were increased significantly in group IV, as compared with group I. The ratio of OX76+CD4+ or OX76+CD8+ T cells to OX76-CD4+ or OX76-CD8+ T cells was greater in group IV, as compared group I. OX76+CD8+ T cells from hepatic allografts in group IV expressed IFN-γ and interleukin (IL)-10, but not IL-2 mRNA. Apoptotic hepatic infiltrates were greater in group IV, as compared to group I. Conclusion: The cytokine profile of donor CD8+ T cells from allografts treated by the intraportal administration of recipient blood is associated with apoptosis of graft-infiltrating cells and the prolonged survival of hepatic allografts in rats. [Copyright &y& Elsevier]

Rad18 plays a crucial role in postreplication repair in both lower eukaryotes and higher eukaryotes. However, regulation of the Rad18 expression in higher eukaryotes is largely unknown. We found that the RAD18 transcript is expressed ubiquitously in various tissues and very highly in the testis in mammals. Although human RAD18 ( hRAD18) transcription levels fluctuate during the cell cycle, being maximal in the late S and minimal in the early G1, the protein levels remain constant throughout the cell cycle. Following UV-irradiation, hRAD18 transcription levels decrease significantly, but Rad18 protein levels change little. The protein levels are maintained at least in part by enhanced translation rates. hRad18 localizes in the nucleus in two forms: a diffused form and a condensed form forming nuclear dots. These nuclear dots disperse rapidly in the nucleoplasm after treatments with various genotoxic agents, resulting in an enhancement of the intranuclear Rad18 concentration of the diffused form. No de novo protein synthesis is required for this process. These results suggest that in higher eukaryotes, the maintenance and dynamic translocation of Rad18 protein is important for postreplication repair. [ABSTRACT FROM AUTHOR]

This study represents a retrospective comparison of video-assisted thoracic surgery (VATS) lobectomy with standard open lobectomy for non-small cell lung cancer (NSCLC). The endpoints of this study include surgical stress as measured by interleukin 6 concentration and patient survival. A retrospective review was performed of 240 consecutive patients with clinical stage IA or IB NSCLC who underwent either VATS lobectomy (n?=?67) or conventional open lobectomy (n?=?173). The amount of blood loss was significantly less in the VATS group (110±75 ml) as compared to 165±90 ml for the open lobectomy group ( P <0.05). A significantly lower incidence of postthoracotomy pain occurred in the VATS group (6.2±4.1 times/3days) than in the open lobectomy group (13.5±5.8 times/3 days, P <0.0001). The postoperative interleukin (IL)-6 serum concentration of was significantly lower in the VATS group (112±43 pg/ml) than that in the open lobectomy group (351±133 pg/ml, P <0.001). There was no statistically significant difference in survival between the VATS and open lobectomy groups. The median follow-up was 42 months in both groups. VATS lobectomy for NSCLC is a reasonable treatment option for selected patients with stage I NSCLC. [ABSTRACT FROM AUTHOR]

Donor dendritic cells (DC) migrate into the recipient spleen after hepatic transplantation. Immunological unresponsiveness to rat hepatic allografts can be induced by prior donor-specific blood transfusion (DST). We investigated homing receptor phenotype and splenic distribution of donor DC after allografting and DST. Immunostaining revealed OX62+ cells in the splenic red pulp of animals receiving pre-transplant DST but only in the white pulp of untreated animals. Most OX62 cells were positive for OX76. There were two subsets of DC in the spleen, CD45RChighOX62+ and CD45RClowOX62+ cells. RT-PCR revealed that CD45RClowOX62+ cells expressed interleukin (IL)-10, while CD45RChighOX62+ cells expressed IL-2 and low levels of IL-10 mRNA. CD45RChighOX62+ cells strongly expressed CCR5 and CCR7, compared with weak expression in CD45RClowOX62+ cells. The Epstein-Barr virus-induced molecule 1 (EBI-1) ligand chemokine (ELC/MIP3β) was expressed mainly within the splenic white pulp. Mucosal vascular addressin-cell adhesion molecule-1 (MAdCAM-1) was expressed in the marginal zone and white pulp, but expression of splenic MAdCAM-1 was down-regulated in DST-treated animals. L-selectin (CD62L), the ligand for MAdCAM-1, was strongly expressed on CD45RChighOX62+ cells but not on CD45RClowOX62+ cells. In conclusion, differential splenic migration of CCR5lowCCR7lowCD62Llow CD45RClow DC expressing Th2-type cytokines is associated with immunological unresponsiveness to rat hepatic allografts. [Copyright &y& Elsevier]

Background: Fatty split-liver and living-related liver transplantation is associated with massive hepatocellular necrosis during acute rejection. Uncoupling protein (UCP)-2 is a potential regulator of energy expenditure and ATP production. We investigated the role of UCP-2 and the effects of a metalloprotease inhibitor, Y-39083, on hepatocellular injury in fatty liver allografts in rats.Materials and methods: Rats were treated for 6 weeks with high-ethanol or isocalic dextrose-containing liquid diets that caused characteristic pericentral lipid accumulation. Alcoholic or nonalcoholic fatty livers from ACI (RT1a) rats were transplanted into LEW (RT1l) rats orthotopically. Hepatic necrosis was determined histologically following liver transplantation. UCP-2 mRNA levels in the hepatic allograft and in primary cultured hepatocytes from fatty liver stimulated by tumor necrosis factor (TNF)-α were determined. Y-39083 was administered to recipient rats continuously at 5 mg/kg/day using an osmotic infusion mini-pump.Results: The acute rejection index on day 5 posttransplant in alcoholic and nonalcoholic fatty donor livers was higher than in lean grafts. Massive hepatocyte necrosis was more prominent in alcoholic than nonalcoholic fatty liver allografts and was not seen in lean allografts. UCP-2 transcripts in both alcoholic and nonalcoholic fatty liver allografts were higher than in lean allografts. Serum TNF-α concentrations in recipient rats with either fatty liver allograft were greater than in animals with lean allografts. In vitro UCP-2 mRNA levels in primary cultured hepatocytes from both alcoholic and nonalcoholic fatty livers increased more after stimulation with TNF-α than those from lean livers. In vitro TNF-α production by Kupffer cells isolated from alcohol-induced fatty liver allografts on day 3 posttransplant was greater than those from lean allografts. Y-39083 significantly reduced serum concentrations of TNF-α and prevented massive hepatocellular necrosis in rats with both alcoholic and nonalcoholic fatty liver allografts.Conclusion: Liver grafts with steatosis up-regulated UCP-2. TNF-α further enhanced UCP-2 transcripts, inducing massive hepatocellular necrosis during acute rejection. Posttransplantation necrosis may be prevented by metalloprotease inhibitors. [Copyright &y& Elsevier]

Calpains form a superfamlly of Ca2+-dependent intracellular cysteine proteases with various isoforms. Two isoforms, µ- and m-calpains, are ubiquitously expressed and known as conventional calpains. It has been previously shown that the mammalian calpains are activated during mitosis by transient increases in cytosolic Ca2+ concentration. However, it is still unknown whether the activation of calpains contributes to particular events in mitosis. With the use of RNA interference (RNAi), we investigated the roles of calpains in mitosis. Cells reduced the levels of m-calpain, but not µ-calpain, arrested at prometaphase and failed to align their chromosomes at the spindle equator. Specific peptidyl calpain inhibitors also induced aberrant mitosis with chromosome misalignment. Although both m-calpain RNAi and caipain inhibitors affected neither the separation of centrosomes nor the assembly of bipolar spindies, Mad2 was detected on the kinetochores of the misaligned chromosomes, indicating that the prometaphase arrest induced by calpain inhibition is due to activation of the spindle assembly checkpoint. Furthermore, when calpain activity was inhibited in cells having monopolar spindles, chromosomes were clustered adjacent to the centrosome, suggesting that calpain activity is involved in a polar ejection force for metaphase alignment of chromosomes. Based on these findings, we propose that activation of m-calpain during mitosis is required for cells to establish the chromosome alignment by regulatlng some molecules that generate polar ejection force. [ABSTRACT FROM AUTHOR]

The immunohistochemical localization of membrane-associated phospholipase A2 (M-PLA2) in normal human pancreases and 30 cases of pancreatic ductal carcinomas was investigated. In pancreatic ductal carcinomas, the immunoreactivity was observed in 25 cases (83%). Among the clinocopathological factors of pancreatic cancer, the incidence of expression of this enzyme is significantly higher in infiltrative type cancers. Furthermore, the expression of M-PLA2 was significantly increased in tumors, which had a larger amount of interstitial tissue. On the other hand, human M-PLA2 added exogenously to the fibroblast cell lines Swiss 3T3 and BALB/3T3 was found to augment their DNA synthesis. The stimulation of DNA synthesis was not affected by treatment with indomethacin. These results suggest that this enzyme could be involved directly in the proliferation of interstitial tissue through its own function. [Copyright &y& Elsevier]

Sahin et al. introduced a new approach, termed serological identification of antigens by recombinant expression cloning (SEREX), for identification of tumor antigens. This approach uses diluted serum from cancer patients to detect prokaryotically expressed cDNA libraries prepared from tumors. Our SEREX study of human pancreatic adenocarcinoma led to the identification of 18 genes. We characterized one of them, heat shock protein 105 (hsp105), which showed over-expression especially in pancreatic ductal and colon adenocarcinoma. Also, our SEREX study of squamous cell carcinoma of head and neck (SCCHN) led to the identification of 19 genes, including three kinds of novel cancer/testis (CT) antigen. These antigens defined by us may be pertinent candidates for cancer-diagnosis and related immunotherapy. [Copyright &y& Elsevier]

We found that EBC-1, a human non-small cell lung cancer (NSCLC) cell line, formed multiple metastases in the lung after subcutaneous inoculation into severe combined immunodeficiency (SCID) mice. SCID mice were injected in the flank with 106 EBC-1 cells. Metastases in the lung were counted in histologic sections and stained immunohistochemically for neutrophil elastase (NE). Solid tumors were palpable in the flank at 3 weeks and grew steadily until 10 weeks. EBC-1 cells formed multiple metastases in the lung at 7 weeks; their numbers increased steadily until 12 weeks in all mice. Immunoreactivity for NE was intense in the metastatic tumor cells. As a marker for circulating tumor cells, human β-actin mRNA was detected in blood by reverse transcriptase–polymerase chain reaction (RT-PCR). Blood samples obtained at 3 weeks after tumor inoculation contained human β-actin mRNA. Our NSCLC EBC-1 pulmonary metastasis model is reliable, technically simple, and predictably results in pulmonary metastasis from early hematogenous spread. This model may be useful for screening potential new drugs, including specific NE inhibitors, for effectiveness against pulmonary metastasis of NSCLC. [Copyright &y& Elsevier]

The proteinase-activated receptor-2 (PAR-2) is a G protein-coupled receptor that is cleaved and activated by trypsin and tryptase. PAR-2 was widely expressed in human pancreatic cancer cell lines and in malignant and nonmalignant human pancreatic tissues. The expression of PAR-2 was found to be higher in the tissues with infiltrative growth pattern than those with expansive growth pattern. Moreover, significantly higher expression of PAR-2 was observed in the tissues that were accompanied with severe fibrosis. Even in the same specimen, the intensity of immunoreactivity tended to be stronger in the part accompanied with severe fibrosis than that accompanied with mild fibrosis. Similarly, the higher expression of PAR-2 was observed in chronic pancreatitis with severe fibrosis than with mild fibrosis. Higher expression of PAR-2 in pancreatic cancer showing infiltrative growth and higher degree of fibrosis may indicate that the activation of PAR-2 is involved in cancer invasion and the induction of fibrosis in human pancreatic cancer. [Copyright &y& Elsevier]

It has been reported that a large number of carcinoma cells are able to produce a variety of cytokines. We also reported that many carcinoma cell lines constitutively produce IL-8 in supernatant cultures. To evaluate the in vivo function of IL-8 of carcinoma cells, we investigated the expression of IL-8 and IL-8 receptors (IL-8R) in human carcinoma cells and the role of IL-8 and IL-8R in the growth of carcinoma cells. IL-8 mRNA was detected in 16 out of 20 (80%) carcinoma cell lines and 20 out of 24 (83.3%) carcinoma tissues by Northern blot analysis. IL-8R mRNA was expressed in 7 out of 11 (63.6%) carcinoma cell lines by reverse transcriptase–polymerase chain reaction (RT-PCR). Neutrophil chemotactic activity in supernatant cultures of carcinoma cell lines correlated with immunoreactive IL-8 concentration. Growth of carcinoma cells was significantly inhibited in the presence of anti-IL-8 antibodies or IL-8R antisense oligonucleotides. These results revealed that IL-8 and IL-8R are expressed in the majority of carcinoma cells, and it suggests that they might play a role in the growth of carcinoma cells.On the other hand, the NF-κB/Rel families are crucial transcriptional factors for regulation of IL-8 in immune and inflammatory responses. To determine if IL-8 production in carcinoma cells is associated with activation of nuclear factor κB (NF-κB), we investigated MCF-7 breast carcinoma cells with 12-O-teradecanoyl-phorbol-13-acetate (TPA) treatment. After TPA treatment, ELISA showed IL-8 production was increasing in culture supernatant. Western blotting revealed that in concert with the decline in cytosolic IκB, NF-κB exhibited similar kinetics of translocation to the nucleus. Electrophoric mobility shift assay (EMSA) also revealed that activation of NF-κB. These findings suggest that activation of NF-κB regulates IL-8 expression in carcinoma cells. [Copyright &y& Elsevier]

Recently, differences in histological diagnosis of colonic neoplasms between Japan and Western countries have been noted. We examined the relationship between the histopathology and immunohistochemical localization of six genetically distinct type IV collagen α chains, the major component of basement membrane (BM), in normal and neoplastic colonic tissues. α1/α2(IV) and α5/α6(IV) chains were observed in normal mucosa and tubular adenomas with mild/moderate atypia. In non-invasive cancers, both α1/α2(IV) chains were linearly stained in the BM of cancer cell nests, while the assembly of α5/α6(IV) chains into the BM was inhibited in a discontinuous or negatively stained pattern. The differential expressions of the type IV collagen α5/α6 chains could be one of the diagnostic markers of the invasiveness of colon cancer. [Copyright &y& Elsevier]

To investigate the biological significance of skin-derived anti-leukoproteinase (SKALP)/elafin in squamous cell carcinoma (SqCC), the immunohistochemical expressions of SKALP/elafin in SqCC (lung 46 and esophagus 34) were studied. The results were compared with the immunohistochemical staining of a proliferating cell nuclear antigen (PCNA) and a TDT-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay in serial sections. The expression of SKALP/elafin had a high incidence in SqCC (lung 82.6% and esophagus 73.5%). In contrast, no expression was observed in uninvolved bronchial epithelium and normal esophagus except for hyperplastic cells. SKALP/elafin expression was located in the cell layer just underneath the cornified envelope of each cell nest, and DNA fragmentation was observed in the same cell layer in which SKALP/elafin was expressed. PCNA expression was observed in the basal layer of each cornified envelope, and both expressions were clearly separated. In lung SqCC, immunoreactive score (IRS) of SKALP/elafin expression correlated significantly with the differentiation and it tended to increase in accordance with the degree of differentiation. In esophageal SqCC, there was also an obvious relationship between the expression of SKALP/elafin and its differentiation. There was no correlation between the expression and other clinicopathological factors. These results suggest that SKALP/elafin is possibly involved in the cell differentiation program, finally leading to keratinization in SqCC of human lung and esophagus. SKALP/elafin may be a beneficial molecular target for detection or even the development of a new therapeutic method against cancer. [Copyright &y& Elsevier]

Our recent study has demonstrated that brain (fetal)-type glycogen phosphorylase (BGP) positive foci (BGP foci) have a vital role in the de novo colorectal carcinogenesis. This paper reviewed the investigation on the genetic alternations in the BGP foci and the clarification of the mechanisms of de novo colorectal carcinogenesis. De novo colorectal carcinomas with invasion into submucosa or superficial muscularis propria were selected from resected specimens. Investigations of the p53, K-ras and APC mutations were performed in the BGP foci, BGP negative colorectal mucosa and de novo colorectal carcinoma. No K-ras mutation was observed in all of the cases. Mutations of p53 and APC were 14 (50.0%) and 9 (32.1%) in de novo colorectal carcinomas, and 11 (39.3%) and 1 (3.6%) in BGP foci, respectively. Both p53 and APC mutations were detected in 8 and 1, p53 mutation alone in 6 and 10, APC mutation alone in 1 and 0 out of 28 de novo colorectal carcinomas and BGP foci, respectively. Our sequential studies propose that the two major pathways, i.e., the p53–APC pathway and the p53 alone pathway in the de novo colorectal carcinogenesis. [Copyright &y& Elsevier]

GC-associated DNA primase (GANP) contains two functionally important regions, the N-terminal RNA/DNA primase and C-terminal MCM3-associated/import domains, which might be involved in DNA replication. Here, we investigated the functions of these regions by transfection studies. In addition to the C-terminal MCM3-binding (Gmap80) region, the RNA/DNA primase region (Gp) possesses an additional MCM3-interaction activity that mediates its translocation between the nucleus and the cytoplasm. Gp with an N-terminal nuclear localization signal sequence (NLS) is present in the nucleus, but Gmap80 with a C-terminal NLS appeared in the cytoplasm. The whole GANP (Ganp) protein is present in both nuclear and cytoplasmic compartments, in a Crm1-dependent manner. Gp associated with MCM3 through the NLS, and MCM3 induced nuclear localization of Ganp. NLS-negative MCM3 does not have such activity. These results suggest that the NLS of MCM3 is involved in nuclear translocation of Ganp. The nuclear-cytoplasm MCM3 translocation was demonstrated in mammalian cells, and over-expression of Ganp, but not of Gp or Gmap80, facilitated this translocation of MCM3 in co-transfectants. Moreover, introduction of Ganp induced DNA synthesis in the transfectants but dominant-negative mutants did not. It is concluded that GANP is translocated from the nucleus to the cytoplasm in association with MCM3 and is involved in DNA synthesis. [Copyright &y& Elsevier]

The presence of RON and its variant isoform in malignant and non-malignant human colonic tissues was investigated by immunohistochemistry using paraffin-embedded sections and RT-PCR analysis followed by direct sequencing of PCR product using RNAs isolated from frozen tissues. In normal colonic mucosa of both human fetus and adult, RON was uniformly expressed in cript cells, especially in the bottom of cripta. On the other hand, the expression was distributed heterogeneously in adenomas and in colon cancer. The expression of RON was significantly related to the degree of differentiation of colon cancer. The RT-PCR analysis of RNA isolated from malignant and non-malignant colonic tissue revealed the presence of two RON mRNA isoforms. Direct sequencing of two major products was revealed to be identical to that of human wild-type RON and a splicing variant of RON transcript which has been reported in human gastric cancer cell line, KATO-III. It is indicated that both wild type RON and its valiant isoform play an important role in regulating the normal function of colonic mucosa such as differentiation and motile activity, and the expression of both wild-type RON and its valiant isoform could be considered to be reduced during malignancy of human colonic mucosa. [Copyright &y& Elsevier]

It is well known that heme oxygenase-1 (HO-1) as well as nitric oxide (NO) synthase expressed in tumor cell line and solid tumor. NO may be beneficial to the tumor growth, not only because enhanced vascular permeability in solid tumor is mediated by NO, but because the l-arginine-dependent NO pathway mediates angiogenic activity and vascular endothelial growth factor (VEGF) induces angiogenesis via formation of NO. However, NO has a powerful cytostatic action on tumor cells by inhibiting DNA synthesis and causing oxidative injury. While it has been suggested that induction of HO-1 may provide an important protective response by cells against oxidative damage, HO-1 expression in tumor cells is strongly enhanced by NO donor. Strong induction of HO-1 is observed in solid tumors after occlusion or embolization of the tumor-feeding artery, indicating that ischemic stress which may involve oxidative stress triggers HO-1 induction. In addition, it is of great importance that an HO inhibitor, zinc protoporphyrin IX, suppressed the tumor growth to a great extent. In conclusion, HO-1 expression in the solid tumor may confer resistance of tumor cells to hypoxic stress as well as to NO-mediated cytotoxicity. [Copyright &y& Elsevier]

Similarities between cancer cells and neutrophil, such as circulating by single cells, attachment to the vascular system of the target and invasion into the target, lead to a hypothesis that cancer cells might produce neutrophil elastase or a very similar protease. Although the production of immunoreactive polymorphonuclear leukocyte elastase (ir-PMN-E) can be measured by enzyme immunoassay (EIA) in human cancer and cancer cell lines, we were having difficulty to detect this at mRNA level.TNF-α stimulation induces neutrophil elastase to a detectable level by RT-PCR in human cancer cell lines. In SUIT2, neutrophil elastase mRNA was detectable from 0.1 ng/ml of TNF-α stimulation which in EBC1 and MCF7 requires 1.0 ng/ml of TNF-α. For PC-3, there was no induction. The RT-PCR direct sequencing method revealed that the induced neutrophil elastase has no mutation.Although the amount of this molecule is low, as neutrophil elastase is a protease, the function in living systems does not depend on their amount but mainly on the activity they display. The result of this study indicates that some of the cancer cells could produce neutrophil elastase by stimulation of TNF-α and also suggests that neutrophil elastase produced by cancer cells is involved in clinical aspects of malignancy under inflammatory conditions. [Copyright &y& Elsevier]

We demonstrated the dynamic aspects of granulocyte activation in rat severe acute pancreatitis which was induced by cerulein and aggravated following lipopolysaccharide (LPS) injection. Pancreatitis induced by cerulein increased intracellular elastase activity of granulocytes in the blood. However, significant systemic cytokinemia was not provoked under such conditions. After induction of severe pancreatitis by LPS, intracellular elastase activity of circulating granulocytes decreased markedly and immediately. The timing of this decrease was concomitant with induction of systemic hypercytokinemia and granulocytes migration into the lung. Overall results provide the following implications: (1) circulating granulocytes are activated by the induction of mild pancreatitis; (2) the activation of granulocytes is mediated by other factors than systemic cytokinemia, such as locally produced cytokine; (3) those priming granulocytes immediately and significantly migrate from the circulation into extravascular space by the induction of endotoxemia; (4) the migration of granulocytes, in turn, may be mediated by systemic cytokinemia. [Copyright &y& Elsevier]

Background: Interleukin-18 (IL-18) production is the main mechanism of hepatic injury in several animal models via further gamma interferon (IFN-γ) release. IL-18 also functions as an inducer of proinflammatory cytokines as TNF-α, which are also shown to induce organ injuries in many severe inflammatory conditions. To clarify the mechanism of hepatic injury developed in acute pancreatitis, plasma concentrations of IL-18 and its related cytokines were analyzed. Patients and methods: Plasma levels of IL-18, IL-12, IFN-γ, TNF-α, soluble TNF receptor (sTNF-R) and soluble Fas ligand (sFas-L) were measured by ELISA. Results: Plasma concentrations of IL-18 in patients with hepatic injury increased during the first few days apparently and remained so to some extent thereafter. However, IFN-γ showed no apparent increase in plasma concentrations. Those of TNF-α and sTNF-R remained increased in the hepatic injury group. Conclusion: These results suggest that IL-18 may act as a pro-inflammatory cytokine, which provokes the cytokine storm and eventually hepatic injury. [Copyright &y& Elsevier]

Objectives: We clarified a distinctive feature of response to surgical stress in elderly patients compared to nonelderly patients. Subjects: Two hundred and ninety-two consecutive patients received elective gastrointestinal surgery. Patients were analyzed for physiological status, cytokine levels, surgical stress parameters and postoperative status of SIRS and complications. Cytokine responses according to patient''s age: Plasma levels of interleukin 6 (IL-6) fluctuated in higher concentrations in elderly patients. Peak plasma levels of IL-6 were significantly high in patients aged over 75 years in total gastrectomy, over 65 years in pancreaticoduodenectomy and over 60 years in major hepatectomy. Mechanisms of hypercytokinemia in elderly patients: Renal dysfunction caused hypercytokinemia in elderly patients. Hypercytokinemia and SIRS: SIRS was more frequently encountered and continued for a long time in elderly patients, giving us an early sign of postoperative complications. Conclusions: In order to prevent postoperative complications in elderly patients, we must shorten status of SIRS due to excessive hypercytokinemia. [Copyright &y& Elsevier]

Reverse transcription-polymerase chain reaction (RT-PCR) technique is extremely useful for sensitive detection of cancer cells, but it requires at least 8 h. If the precise information on the cancer spread based on RT-PCR is available during surgery, we can give the optimal therapy to each patient during operation. We aimed to establish the ultra-rapid procedure of RT-PCR for application to the intra-operative diagnosis. We have established a one-step real-time RT-PCR using LightCycler with hybridization probes, in combination with automated mRNA extraction on MagNA Pure LC. The method can be completed in only 70 min after sampling and has a comparable sensitivity to conventional RT-PCR and/or two-step real-time RT-PCR with a wide (10–106) dynamic range. When this method was applied to the intra-operative detection of free cancer cells in peritoneal lavage, the results were available during operation in all cases. We could detect cancer cells more effectively compared to conventional cytology and RT-PCR. This protocol is easily accomplished, delivers rapid, sensitive and reproducible results, and is therefore applicable to intra-operative diagnosis. [Copyright &y& Elsevier]

Purpose: We aimed to clarify the possibility of hematogenous spreading of cancer cells by microwave coagulation therapy (MCT) and radio-frequency ablation (RFA) for liver cancers. Molecular marker: Carcinoembryonic antigen (CEA) mRNA and melanoma antigen-3 (MAGE-3) mRNA were selected as a genetic marker. PCR method: Blood samples were collected before, during and just after the treatments. Genetic markers were examined using LightCycler reverse transcriptase–polymerase chain reaction (RT–PCR) assay with SYBR Green I. The samples with both positive markers were defined as genetic positive. Molecular metastasis and recurrence: We evaluated peripheral blood samples of 70 patients with liver tumors; 64 hepatocellular carcinomas (HCCs), 1 cholangiocellular carcinoma (CCC), and 5 metastatic liver tumors (MLTs). Genetic positive rates in RT–PCR assay before treatment were 42.2% in HCC, 0% in CCC, and 20% in MLT. Among genetic negative patients before operation, 45% of patients in MCT and 29% in RFA changed to genetic positive as a result of the procedures. Recurrence rates were 55% and 50% in MCT and 43% and 88% in RFA of the patients assessed negative to negative and negative to positive, respectively. Conclusion: Local ablation therapies may promote hematogenous seeding of cancer cells. [Copyright &y& Elsevier]

The established ultra-rapid quantitative RT-PCR system for intraoperative detection of intraperitoneal free cancer cells, the elucidated cause of peritoneal recurrence after curative operation for patients with non-serosa-invasive gastric cancer, and the extensive intraperitoneal lavage (EIPL) therapy for the prophylaxis of peritoneal recurrence from the results of evaluation using the ultra-rapid quantitative RT-PCR system were reviewed. We established ultra-rapid RT-PCR protocol that enables diagnosis of intraperitoneal cancer spread about 70 min, using LightCycler method in combination with automated mRNA extractor. Both the carcinoembryonic antigen (CEA) and cytokeratin (CK)20 messenger RNA (mRNA) in intraperitoneal lavages after lymph node dissection were identified in three (14.2%) and four (26.7%) patients with submucosal (SM) and muscularis propria (MP) tumors, respectively. Although peritoneal metastasis occurs in some patients with early gastric carcinoma, the reasons have not been fully established. These results clarified that peritoneal metastasis after operation for non-serosal-invasive gastric cancer was caused by opening lymphatic channels during lymph node dissection allowing the spread of viable cancer cells into the peritoneal cavity. Furthermore, EIPL method was performed in five cases with serosa-invasive (SE) gastric carcinoma, and its efficacy was evaluated by the quantitative RT-PCR. The quantitative RT-PCR demonstrated that EIPL reduced free cancer cells from 3.8×105±1.4×105 to 2.8±1.5 cells/100 ml of lavage by from six to eight washes, and they disappeared after the seventh to ninth wash. In conclusion, an ultra-rapid quantitative RT-PCR system for intraoperative detection of intraperitoneal free cancer cells was established for clinical use. It is suggested that the combination with the novel rapid detection system with the intraoperative therapy of EIPL can be a useful prophylactic strategy for peritoneal metastasis from gastric carcinoma. [Copyright &y& Elsevier]

Pancreatic secretory trypsin inhibitor (PSTI) is a potent natural inhibitor of trypsin. We proposed a hypothesis: if the function of PSTI is impaired by its genetic mutation, trypsin may easily promote autodigestion causing pancreatitis, and performed mutational analysis of PSTI gene in patients with pancreatitis. Two exonic mutations (N34S and R67C) were suggested to be associated with the predisposition to pancreatitis. N34S mutation was co-segregated with two intronic mutations, IVS1-37T>C and IVS3-69insTTTT. Although we analyzed the function of recombinant N34S protein, we could not demonstrate the loss of function of this protein. Intronic mutations rather than N34S itself (IVS1-37T>C+N34S+IVS3-69insTTTT complex) may be associated with the decreased function of PSTI. Alternatively, increased digestion of N34S in vivo may be applicable. As for R67C, the conformational alteration of the protein by forming intramolecular or intermolecular disulfide bonds with 67Cys was strongly suggested. These results, along with the brand new findings in PSTI knockout mice, suggest that genetic mutation of PSTI is one of the important mechanisms for predisposition to pancreatitis by lowering the trypsin inhibitory function. [Copyright &y& Elsevier]

With the global pandemic of hepatitis B and C infections, the incidence of Hepatocellular carcinoma (HCC) is rapidly increasing world wide. We identified glypican-3 (GPC3), a novel oncofetal gene over-expressed specifically in human HCC, as based on data of cDNA microarrays. As GPC3 is a GPI-anchored membrane protein and could be secreted, we attempted to detect secreted GPC3 protein in sera from HCC patients using Western blotting and ELISA. GPC3 protein was positive in sera of 40.0% (16/40) of HCC patients, and negative in sera from subjects with liver cirrhosis (LC) (0/13), chronic hepatitis (CH) (0/34), and healthy donors (0/60). All subjects were Japanese. Although 12 of 40 HCC patients were negative for both α-fetoprotein (AFP) and PIVKA-II well known tumor markers of HCC, four of these were GPC3-positive in the sera. We also observed vanishing GPC3 protein in the sera of three patients after the surgical treatment for HCC. On the other hand, immunohistochemical analysis revealed that HCC expressed GPC3 protein in all 14 HCC patients tested. In conclusion, GPC3, as defined in this study was shown to be a useful tumor marker for cancer-diagnosis for large numbers of patients with HCC. [Copyright &y& Elsevier]

Background. Mutations in the pancreatic secretory trypsin inhibitor (PSTI) gene have been reported in patients suffering from chronic pancreatitis. The aim of the present study was to investigate whether similar gene mutations are present in familial and juvenile pancreatitis patients in Japan. Methods. All four exons of the PSTI gene and their flanking intronic sequences were amplified by polymerase chain reaction and sequenced for 37 familial pancreatitis patients (24 families) and 15 juvenile pancreatitis patients, distributed throughout Japan. Results. Three types of exonic mutation in the PSTI gene were observed. The N34S mutation was found in six familial pancreatitis patients (three families) and in one juvenile pancreatitis patient, and the R67C mutation was found in one familial pancreatitis patient and one juvenile pancreatitis patient. We also found a 272C>T mutation in the 3' untranslated region of exon 4 in one familial pancreatitis patient and four juvenile pancreatitis patients. It should be noted that the N34S mutation was cosegregated with two intronic mutations, specifically, IVS1-37T>C and IVS369insTTTT. Conclusions. The same set of N34S mutations (N34S + IVS1-37T>C + IVS3-69insTTTT) that exists in other countries was found in the PSTI gene in Japanese familial and juvenile pancreatitis patients. Another unique mutation (R67C) was also observed in two patients; 272C>T was suggested to be a normal polymorphism. [ABSTRACT FROM AUTHOR]

Study Objectives: To establish a clinically relevant animal model of pulmonary metastases of human non-small cell lung carcinoma (NSCLC) cells in severe combined immunodeficiency (SCID) mice, which can be used for repetitive investigations, so as to improve our understanding and management of the cellular and molecular mechanisms of human lung cancer metastases.Methods and Results: SCID mice subcutaneously injected in the flank with 1 x 10(6) EBC-1 cells derived from human lung squamous cell carcinoma were killed weekly for examination until 12 weeks after tumor inoculation. The biological characteristics of implanted tumors and their metastatic foci were investigated by hematoxylin-eosin staining and immunostaining for neutrophil elastase (NE). Three weeks after ectopic implantation, EBC-1 cell lines formed a tumor at the inoculation site and grew steadily to show a plateau at 10 weeks. EBC-1 cells formed multiple metastases in the lung 7 weeks after tumor inoculation; their numbers increased steadily until 12 weeks in all mice. Immunoreactivity for NE was intense in the metastatic tumor cells. Then, to establish the primary tumor amputation/pulmonary metastasis model and to evaluate how primary tumor amputation influences the development of pulmonary metastases at the cellular and molecular level, excision was performed before (3 weeks and 5 weeks after inoculation) and after (7 weeks and 9 weeks after inoculation) formation of lung metastases. When the primary tumor was excised 3 weeks after tumor inoculation, all mice had pulmonary metastasis at 12 weeks after inoculation. Blood samples obtained at 3 weeks after tumor inoculation contained human beta-actin messenger RNA, which represents circulating tumor cells.Conclusion: Our NSCLC EBC-1 pulmonary metastasis model is reliable, technically simple, and predictably results in pulmonary metastasis from early hematogenous spread. This model may be useful for elucidating the mechanism of pulmonary metastasis in human lung cancer, and testing anti-metastatic efficacy of therapeutic agents in vivo. [ABSTRACT FROM AUTHOR]

Improved delivery of macromolecular drugs to solid tumor is known as the enhanced permeability and retention (EPR) effect of macromolecular drugs and lipids. We report here that a prostaglandin I 2 (PGI 2 ) analogue induces enhancement of tumor-selective drug delivery, while it decreases tumor blood flow, in a rat tumor model (AH136B). Beraprost sodium (BPS) is an analogue of PGI 2 that is more stable than parental PGI 2 in vivo ( t 1/2 for BPS is >1 h vs. a few seconds for PGI 2 ). Thus, BPS was administered to tumor-bearing rats to examine its effect on tumor vascular permeability as well as tumor blood flow. The amount of extravasation of the Evans blue-albumin complex in tumor tissue increased from two to three times, whereas tumor blood flow decreased almost 70%, in the group treated with BPS at 7 μg/kg compared with controls. Tissue blood flow of normal organs such as the kidney and the liver did not change to a significant extent. These findings establish a new role for BPS, not only in enhancing macromolecular drug delivery, but also in reducing the blood supply to tumor tissues. [ABSTRACT FROM AUTHOR]

Background. We hypothesized that mutation of the pancreatic secretory trypsin inhibitor (PSTI) gene may promote a predisposition to pancreatitis, possibly by reducing the inhibition of trypsin activity. Based on this hypothesis, we performed a biochemical analysis of recombinant PSTI protein. Methods. The trypsin inhibitory activity of the recombinant protein was analyzed. The activity of PSTI protein with a point mutation of the most common type: [sup 34]Asn (AAT)-to-Ser (AGT)(101A>G N34S: N34S) in exon 3, was compared with that of the wild type. Results. The function of N34S PSTI remained unchanged under both the usual alkaline and acidic conditions compared with the wild-type PSTI. The calcium concentration did not affect the activity of recombinant PSTI. The trypsin susceptibility of the N34S protein was not increased either. Conclusions. Mechanisms other than the conformational change of PSTI associated with amino-acid substitution, such as abnormal splicing, may underlie the predisposition to pancretitis in patients with the N34S mutation. [ABSTRACT FROM AUTHOR]

Nonsteroidal anti-inflammatory drugs (NSAIDs) have a preventive effect against colorectal cancer. Although inhibition of cyclooxygenase-2 plays a crucial role in the suppression of tumors, precise mechanisms of their action remain to be disclosed. To identify genes involved in the growth-suppressive effect of NSAIDs, we utilized cDNA microarray containing 23,040 genes and analyzed time-dependent alteration of gene expression in response to sulindac or aspirin in NSAIDs-sensitive SW480 and SW948 colon-cancer cells as well as in relatively resistant SNU-C4 cells. Consequently we identified 112 genes with commonly altered expression by sulindac and 176 with commonly altered expression by aspirin in the three lines. Addition of sulindac and that of aspirin altered expression levels of 130 and 140 genes, respectively, in SW480 and SW948 cells but not in SNU-C4 cells. These data may lead to a better understanding of growth-suppressive effects on colonic epithelium, and may provide clues for identifying novel therapeutic and/or preventive molecular targets of colon cancer. [Copyright &y& Elsevier]

In this study, we examined the relationship between the histopathological grade and immunohistochemical localization of six genetically distinct type IV collagen α chains, the major component of basement membrane (BM), in normal and neoplastic colorectal tissues. In the normal colorectal mucosa, α1/α2(IV) and α5/α6(IV) chains were stained in all epithelial BM. However, α3/α4(IV) chains were restrictively immunostained in the BM of the apical surface epithelium. Similar immunostaining profiles for α1/α2(IV) and α5/α6(IV) chains were observed in tubular adenomas with mild/moderate atypia. However, in intramucosal carcinomas, both α1/α2(IV) chains were linearly stained in the BM of cancer cell nests, while the assembly of α5/α6(IV) chains into the BM was inhibited in a discontinuous or negatively stained pattern. The normal colorectal mucosa forms a second network of BM composed of α5/α6(IV), partly α3/α4(IV) chains, in addition to the classic network of α1/α2(IV) chains. The differential immunohistochemical localization of the type IV collagen α5/α6 chains could be one diagnostic marker for the invasiveness of colorectal cancer. [ABSTRACT FROM AUTHOR]

Purpose: We hypothesized that mutations in the pancreatic secretory trypsin inhibitor (PSTI) gene could promote autodigestion, leading to acute or chronic pancreatitis. Our investigation involved mutation analysis of the PSTI gene in patients with acute or chronic pancreatitis.Methods: Mutation analysis for the PSTI gene was performed in patients with acute or chronic pancreatitis. Unrelated healthy volunteers and family members of a chronic pancreatitis patient with point mutations in the PSTI gene were also analyzed.Results: Two types of single-point mutation in the PSTI gene were observed in one patient with chronic pancreatitis: 34Asn (AAT)-to-Ser (AGT) (101 A > G N34S: N34S) in exon 3, and 67Arg (CGC)-to-Cys (TGC) (199 C > T R67C: R67C) in exon 4. No mutations with amino-acid substitution were found in other patients or in the volunteer group. In the patient with the PSTI gene mutations, no additional mutations were observed in the cationic trypsinogen gene. The family study revealed that the mother and a maternal uncle were homozygotes for the N34S mutation, while the father and brother were compound heterozygotes for the N34S and R67C mutations. The uncle (N34S/N34S) showed clinical manifestations of pancreatitis, but the other family members did not.Conclusions: The N34S mutation may cause a predisposition to pancreatitis, with incomplete penetrance. However, with the limited information available, it is not known whether the R67C mutation promotes pancreatitis. [ABSTRACT FROM AUTHOR]

Reports the involvement of DNA-primase germinal center-associated nuclear protein (GANP) in regulation of cell proliferation of antigen-driven B cells in germinal centers. Effect of over expressed GANP in cell cycling; Need of GANP DNA-primase activity in organs with extraordinary rapid cell proliferation; Phosphorylation of GANP DNA-primase.