AURKA is one of the downstream targets of MAPK1/ERK2 in pancreatic cancer.

DUSP6/MKP-3, a specific inhibitor of MAPK1/ERK2, frequently loses its expression in primary pancreatic cancer tissues. This evidence suggests that constitutive activation of MAPK1 synergistically induced by frequent mutation of KRAS2 and the loss of function of DUSP6 plays key roles in pancreatic carcinogenesis and progression. By profiling of gene expressions associated with downregulation of MAPK1 induced by exogenous overexpression of DUSP6 in pancreatic cancer cells, we found that AURKA/STK15, the gene encoding Aurora-A kinase, which plays key roles in cellular mitosis, was among the downregulated genes along with its related genes, which included AURKB, TPX2 and CENPA. An association of expression and promoter activity of AURKA with MAPK activity was verified. Knockdown of ETS2 resulted in a reduction of AURKA expression. These results indicate that AURKA is a direct target of the MAPK pathway and that its overexpression in pancreatic cancer is induced by hyperactivation of the pathway, at least via ETS2.

Immunohistochemical classification of the localization of laminin in the thickened bronchial epithelial basement membrane of deceased bronchial asthma patients.

To ascertain histological changes in the basal lamina of the bronchial epithelial basement membrane in patients with severe bronchial asthma, an immunohistochemical study was conducted in 43 patients who died of bronchial asthma. Antibodies against laminin, a component of the lamina lucida, were utilized. The results revealed various patterns for immunoreactivity to laminin in the thickened basement membrane layer. We were able to classify these reactivities into four patterns. In Pattern A, laminin reactions branched vertically in relation to the thickened basement membrane layer. In Pattern B, laminin reactions formed lines along the lower margin of the thickened basement membrane layer. In Pattern C, laminin reactions formed lines along the upper margin of the thickened basement membrane layer. Finally, in Pattern D, no laminin reactions were observed. In addition, relationships between immunohistological characteristics of laminin and findings such as epithelial cell shedding, basal cell proliferation and basement membrane layer thickening were investigated. In many Pattern A patients, epithelial cell shedding was observed, but goblet cell hyperplasia and basal cell proliferation were barely detectable. Conversely, in numerous Pattern D patients, epithelial cell shedding was barely seen, but goblet cell hyperplasia and basal cell proliferation were marked. Hence, Patterns A and D were on opposite ends of the spectrum of morphological characteristics associated with severe bronchial asthma. In Patterns B and C, laminin reactions formed lines along the lower and upper margin of the thickened basement membrane layer, respectively. However, no marked differences existed in epithelial cell shedding and basement membrane layer thickening. The present study is thus the first to clarify that laminin reactions in the thickened basement membrane layer vary, and this feature is unique to the bronchi of patients with severe bronchial asthma.

Living-related partial liver transplantation for decompensated hepatitis B without reactivation of hepatitis B in the following 30 months.

We report a case of living-related partial liver transplantation for decompensated hepatitis B without reactivation of hepatitis B in the following 30 months, and we analyze the factors that indicate a favorable prognosis for transplantation. The 42-year-old female patient received continuously administered lamivudine before transplantation, and hepatitis B virus immunoglobulin (HBIG) from the anhepatic phase to the present. Currently, she shows a normal aminotransferase level and is negative for hepatitis B surface antigen and hepatitis B virus (HBV) DNA by polymerase chain reaction amplification. Sequence analysis was performed. The entire precore/core region and part of the polymerase region of HBV were sequenced by a direct sequencing method after polymerase chain reaction. No specific mutation was found in these regions. These observations show that the key factors in the long-term successful treatment of this patient appear to be the combination therapy of lamivudine and HBIG that the patient received from around the time of the transplantation. Furthermore, the lack of specific mutations, including lamivudine resistant-mutations, is likely to represent an additional factor in the effectiveness of this treatment.

Transpapillary intraductal US prior to biliary drainage in the assessment of longitudinal spread of extrahepatic bile duct carcinoma.

BACKGROUND: The utility of intraductal US via the transpapillary route prior to biliary drainage in the assessment of longitudinal extension of extrahepatic bile duct carcinoma was investigated. METHODS: In 19 patients with extrahepatic bile duct carcinoma who underwent surgical resection, an ultrasonic probe (diameter, 2.0 mm; frequency, 20 MHz) was inserted into the bile duct via the transpapillary route prior to biliary drainage. Longitudinal cancer extension along the bile duct was prospectively determined and compared with the histologic findings in the resected specimens. RESULTS: Results on the hepatic side were as follows: Intraductal US demonstrated more extensive longitudinal cancer spread than cholangiography in 9 of 19 patients with one instance of overdiagnosis. The accuracy of intraductal US in assessing the extent of spread (84%) was superior to that of cholangiography (47%) (p < 0.05). Results on the duodenal side were as follows: In patients with suprapancreatic bile duct cancer (n = 14), intraductal US demonstrated more extensive longitudinal cancer spread than cholangiography in 8 of 14 patients. The accuracy of intraductal US in assessing the extent of the spread (86%) was superior to that of cholangiography (43%) (p < 0.05). CONCLUSIONS: Transpapillary intraductal US prior to biliary drainage is useful in demonstrating longitudinal extension of bile duct cancer. However, the surgical margins were inaccurate in some patients.

Identification and characterization of a novel gene that is upregulated in leukaemia cells by nitric oxide.

Nitric oxide (NO) inhibits growth and induces differentiation in acute myeloid leukaemia (AML) cells. To identify genes associated with these processes, we studied the effect of NO on AML gene expression using the technique of Representational Difference Analysis. Exposure of HL-60 cells to the NO donor DETA-NO for 24 h induced the expression of a novel gene that was named rno (regulated by nitric oxide). Treatment of HL-60 cells with dimethyl sulphoxide induced expression of rno, but treatment with Vitamin D3 or all-trans retinoic acid did not. Upregulation of rno by NO was cGMP independent. Northern blot analysis indicated that constitutive expression of the novel gene was limited to leucocytes. Three isoforms of rno were identified. An rno cDNA clone was obtained by screening a human leucocyte library. The nucleotide sequence of the open reading frame shared significant homology with that of the human ribonuclease/angiogenin inhibitor (RI). The predicted amino acid sequence indicated that, like RI, rno is leucine and cysteine rich and is comprised of a series of repetitive elements (leucine-rich repeats) that may mediate macromolecular interactions. Enhancement of expression of rno may be a component of the process by which differentiation and growth inhibition of leukaemia cells is induced by NO.

Utility and limitations of intraductal ultrasonography in distinguishing longitudinal cancer extension along the bile duct from inflammatory wall thickening.

BACKGROUND: We wanted to distinguish wall thickening caused by cancer extension from that caused by inflammation after placing a biliary catheter on intraductal ultrasonography (IDUS). METHODS: We studied 51 patients with biliary tract malignancies who had undergone placement of biliary drainage catheters before IDUS. IDUS was performed from a transhepatic (n = 34) or transpapillary (n = 17) route with a thin-caliber ultrasonic probe (2.0 mm in diameter, 20-MHz frequency). At the hepatic side of the tumor, the thickness, asymmetry, outer margin, inner margin, and internal echoes of the bile duct wall were reviewed prospectively and correlated with the histologic findings of the surgically resected specimens in all cases. RESULTS: When IDUS showed wall thickening in a semicircular fashion, notched outer margin, rigid inner margin, papillary inner margin, and heterogeneous internal echoes, each finding had a positive predictive value for diagnosing cancer extension (100%, 100%, 83%, 100%, and 90%, respectively). When these factors were used as the diagnostic criteria of cancer extension, IDUS accurately demonstrated suitable surgical margins in 76% of all patients and 71% of patients with bile duct carcinoma. CONCLUSION: Wall thickening in a semicircular fashion, notched outer margin, rigid or papillary inner margin, and heterogeneous internal echoes are specific for cancer extension. However, surgical margins can be inaccurately assessed in some patients.

Biliopancreatic fistula associated with intraductal papillary-mucinous pancreatic cancer: institutional experience and review of the literature.

Intraductal papillary-mucinous tumour is clinicopathologically characterized by papillary growth and mucin production within the pancreatic duct system. The category includes a wide range of dysplasia, ranging from adenoma to carcinoma, the latter designated as intraductal papillary-mucinous cancer. In general, the tumor renders a favorable prognosis after complete resection. However, intraductal papillary-mucinous tumor with overt invasion outside the gland has been reported to have a poor prognosis, as is the case with the usual type of duct cell cancer of the pancreas. We experienced two cases of intraductal papillary-mucinous cancer with obstructive jaundice due to impaction of thick mucus protruding from the pancreas via a "spontaneous" biliopancreatic fistula. Preoperative examinations of both patients showed a large intraductal papillary-mucinous tumor in the head of the pancreas with fistula formation between the intrapancreatic portion of the common bile duct and the main pancreatic duct. Histopathological investigation of the two resected specimens suggested that the fistula may not have developed from invasion by papillary or tubular adenocarcinoma, but from compression and destruction of the intercalating tissues by abundant mucinous secretion. The first patient died of peritoneal carcinomatosis with clinicopathologic features of pseudomyxoma peritonei 6 years after surgery. The second patient is alive and has been well for 2 years postoperatively. Review of the world literature showed that half of the patients with intraductal papillary-mucinous cancer plus biliopancreatic fistula had no stromal invasion around the fistula, indicating that the fistula might have been caused by mechanical pressure. However, the other half of the cases did have stromal invasion around the fistula. Two-thirds of these cases, including our own patients, had foci of mucinous carcinoma in the stroma around the fistulization, implying that mucinous lakes in the stroma may have served as part of the "waterway" from the pancreatic duct to the bile duct, assisted by increased pressure by mucus production. Since intraductal papillary-mucinous cancer with biliopancreatic fistula has a comparatively favorable prognosis, surgical resection should be considered.

Inhibition of Epstein-Barr virus (EBV)-specific CD8+ cytotoxic T lymphocyte (CTL) activity by soluble HLA class I in vitro.

In the present study, the effects of soluble HLA (sHLA) class I molecules against EBV-specific CTL were examined. Two different sources of sHLA class I, either bioengineered spliced form of HLA-B7 (sB7) or natural production from EBV-transformed B cells (natural sHLA), were added during the induction of CTL or incubated with MHC-restricted CD8+ CTL, which were selected by immunobeads just before testing for their cytotoxic activity. Both sB7 and natural sHLA class I blocked the generation of CD8+ CTL and also inhibited the cytotoxic activity of established CTL in a dose-dependent manner. In both ways, natural sHLA class I was effective in 10-fold lower concentrations compared with sB7. The inhibitory effect did not require a sharing of the HLA allotypes between sHLA and the CTL. CTL, after being treated with sHLA, underwent apoptosis, which was considered here as the main mechanism.

Xenotransplantation of the liver.

Xenotransplantation of the liver, in its broadest conception, might involve the transplantation of an intact organ or xenogeneic hepatocytes, or the use of an intact xenogeneic liver or cells as an ex vivo "device." The indications for xenotransplantation include not only hepatic failure but also, potentially, the treatment of metabolic diseases. The hurdles to xenotransplantation include immune, physiologic, and infectious complications. New information and progress in experimental systems are bringing xenotransplantation closer to clinical application.

Delayed hyperacute xenograft rejection in porcine to canine fetal liver transplantation.

Fetal tissues are generally considered to express weaker antigenic cell-surface molecules than adult tissues. We have reported that transplantation of porcine fetal liver tissue (fragments) is useful for acute and chronic hepatic failure in rats. We further investigated, in the present study, whether transplantation of a porcine fetal liver has the advantage of delayed hyperacute xenograft rejection (HAR) in comparison with that of an adult liver. Porcine fetal liver heterotopically transplanted into dogs was compared. Haematoxylin-eosin (HE) and immunohistochemical studies using IgM, C3, IgG antibodies were performed in serial biopsies of the liver grafts. Lectin binding to target antigen epitopes on pig and dog tissues was studied by flow cytometry. Carbohydrate expression on the liver was also studied by immunohistochemistry. The macroscopic and HE section findings indicate that HAR started 15 min postgraft in fetal and adult liver grafts. Thereafter, vascular changes and parenchymal damage progressed more rapidly in the adult grafts. The final HAR time in adult liver transplantation was determined to be 60 min, while it was determined to be 180 min in fetal liver transplantation. IgM, C3 and IgG were deposited more strongly in the adult grafts than in the fetal grafts up until 60 min after xenografting. Phaseolus vulgaris erythroagglutinin lectin competitively blocked dog sera binding to porcine PBLs. The fetal liver expressed oligosaccharide at a significantly lower level than the adult liver. We conclude that porcine fetal liver xenografts had a significantly delayed HAR.

Beneficial effects of immunoisolated fetal and neonatal pig liver fragments on acute liver failure in a large animal.

Xenogeneic cell (fragment) transplantation may be used as an interim therapy until the organ allotransplanation. Immunologic rejection, however, constitutes the major hurdle. To overcome this problem, "xeno" fetal and neonatal liver fragments (FLF, NLF) were encapsulated into separate micropore devices that protect them from immunological attack by the recipient. The FLF or NLF were then transplanted into beagles with hepatic failure to observe their biological effects. In Experiment 1 (n = 5) beagles were injected IV with D-galactosamine (D-gal, 1.0 g/kg) on day 0 and then received FLF grafts (0, 0.3, 0.8, 1.0, 2.0 g/kg). In Experiment 2 (n = 6) beagles received NLF grafts (1.8 g/kg) and on the following day were injected with D-gal (1.0 g/kg). In Experiment 1 only the high dose of xeno-FLF (2.0 g/kg) decreased the elevated ALT (GPT) and T. Bil. levels. Histologic examination showed that some of the hepatocytes of the host liver survived only in the high-dose graft. In Experiment 2, at 36 and 48 h after D-gal injection, the transplanted group had a significantly lower AST (GOT) level than the control. The grafted NLF survived for 14 days, according to histologic examinations. Thus, encapsulated FLF and NLF xenotransplantation can prevent liver dysfunction in a large animal hepatic failure model.

Prediction of the histologic type of bile duct cancer by using intraductal ultrasonography.

BACKGROUND: Patients with papillary adenocarcinoma survive longer than do patients with other histologic types of bile duct tumors. We evaluated the usefulness of intraductal ultrasonography (IDUS) for predicting the histology. METHODS: Preoperative tumor assessment was performed by using IDUS through a percutaneous tract or the transpapillary route in 37 patients with extrahepatic bile duct cancer. In 30 of 37 patients, imaging results were compared prospectively with histologic findings in resected specimens. Probes 2.0 mm in diameter and 20 MHz in frequency were mainly used. When IDUS showed a "narrow-based polypoid pattern" or a "papillary surface pattern," the patients were judged as having papillary adenocarcinoma. RESULTS: The accuracy, sensitivity, and specificity of IDUS in predicting papillary adenocarcinoma were 90%, 89%, and 90%, respectively. When intraductal ultrasonography showed a papillary surface pattern or a narrow-based polypoid pattern, lymph node metastases and perineural invasion were rarely seen when compared with other patients with bile duct cancer (p < 0.05). CONCLUSION: IDUS is useful for assessing the histologic type of bile duct cancer.

[Systemic lupus erythematosus presenting as fulminant thrombotic thrombocytopenic purpura].

A 20-year-old woman visited a nearby hospital because of sudden, severe, and unusual genital bleeding. She also exhibited severe anemia and thrombocytopenia. In transit to our hospital, the patient suddenly suffered cardiac arrest and died soon thereafter despite immediate blood transfusion and therapeutic intubation. Thrombotic thrombocytopenic purpura (TTP) was initially diagnosed at autopsy due to the observation of numerous fragmented erythrocytes in peripheral blood, evidence of hemolysis, and thrombotic microangiopathy in multiple organs. In addition, histopathologic and serologic findings disclosed an association with systemic lupus erythematosus (SLE). Test for anticardiolipin antibody was positive, and hemophagocytic findings were detected in lymph node specimens. Reports of TTP in association with SLE have been increasing in recent years. However, the mechanisms correlating these two illnesses have not been identified. We speculated that the rapid clinical course in this case was attributable to TTP that had been provoked by endothelial microangiopathy due to SLE, and moreover, the fact that the patient's general condition had been seriously complicated by excessive menstrual bleeding.

Limitation of cholangiography in assessing longitudinal spread of extrahepatic bile duct carcinoma to the hepatic side.

BACKGROUND: Preoperative assessment of longitudinal spread of bile duct carcinoma (BDC) to the hepatic side remains a difficult problem for diagnostic imaging. METHODS: We studied the accuracy of cholangiography in assessing BDC. In 54 patients with extrahepatic bile duct cancer, cholangiographic findings were compared retrospectively with the histological findings of the resected specimens. RESULTS: Histological examination of specimens indicated longitudinal spread of the tumour to the hepatic side in 22 of 54 patients. The accuracy of cholangiography in assessing the extent of the longitudinal spread was only 34/54 (63%). When the cholangiographic images showed a main tumour with a collapsed edge, there was a significantly higher frequency of longitudinal spread compared with tumours with sharp edges (P< 0.05). In contrast, the accuracy of mapping biopsy under percutaneous transhepatic cholangioscopy (n=24) was 83%. CONCLUSIONS: Cholangiography cannot accurately assess the extent of the longitudinal spread of bile duct cancer. When cholangiographic images show a tumour with a collapsed edge, preoperative or intraoperative histological examination is essential to determine a suitable surgical line.

Paroxysmal nocturnal hemoglobinuria: analysis of the effects of mutant PIG-A on gene expression.

Compelling evidence indicates that mutations in PIG-A are necessary for the development of paroxysmal nocturnal hemaglobinuria (PNH), however, it is unclear why mutant PIG-A stem cells have a selective advantage. Further, multiple, discrete PIG-A mutations have been detected in the peripheral blood and bone marrow of patients with PNH, but the contribution of the different mutant clones to hematopoiesis is variable. This observation implies that factors in addition to mutant PIG-A influence the proliferative properties of the abnormal cells. To investigate the etiology of the selective advantage and the clonal dominance in PNH, gene expression in cells with mutant PIG-A was analyzed. Representational difference analysis was used to compare the pattern of cDNA expression between a human lymphoblastoid cell line with mutant PIG-A and its wild-type counterpart. These experiments demonstrated that the pattern of gene expression was different between the two cells lines in that the PIG-A mutant cells failed to express antiquitin mRNA. Transfection of the mutant cells with normal PIG-A restored expression of glycosyl phosphatidylinositol anchored proteins but not antiquitin. These experiments demonstrate that differences in the pattern of gene expression can occur independent of the PIG-A mutation. Depending upon the functional properties of the involved genes, these differences could influence the proliferative properties of PIG-A mutant cells and contribute to the selective advantage and clonal dominance that characterize PNH.

Rapid death following papillary serous carcinoma of the endometrium without myometrial invasion.

A 72-year-old woman was diagnosed with a stage I endometrial carcinoma, and total hysterectomy, bilateral salpingo-oophorectomy, and systematic pelvic and paraaortic lymphadenectomy were performed. Postoperative pathological examination determined that the tumor was confined to the endometrium, with no myometrial invasion or lymph-vascular involvement; histological examination revealed a papillary serous carcinoma of the endometrium. Peritoneal washing cytology during surgery revealed an adenocarcinoma. Despite postoperative adjuvant chemotherapy, early recurrence resulted in death 13 months after surgery. In the absence of myometrial invasion and lymph-vascular involvement, the data suggest that peritoneal washing cytology may serve as a prognostic factor in papillary serous carcinoma of the endometrium.

A novel method of endoscopic mucosal resection using sodium hyaluronate.

BACKGROUND: Saline-assisted endoscopic mucosal resection is an established therapeutic method. However, it is sometimes difficult to maintain a desired level of tissue elevation after injection of saline. Therefore we decided to use a mucinous substance such as sodium hyaluronate instead of saline. METHODS: Two resected porcine stomachs and five dogs were used for the study. The elevations, made by submucosal injections of sodium hyaluronate, were compared with those produced with normal saline. Sodium hyaluronate-assisted mucosal resections were compared with the saline-assisted resections. RESULTS: Mucosal elevations created by submucosal injections of sodium hyaluronate remained for a longer time with a clearer margin compared to those made by saline injection. Endoscopic mucosal resections were performed safely with the assistance of sodium hyaluronate. CONCLUSIONS: Use of sodium hyaluronate instead of saline for endoscopic mucosal resection could make the procedure easier and more reliable.

Study of the relation between fluid distribution change in tissue and impedance change during hemodialysis by frequency characteristics of the flowing blood.

Erythrocyte orientation and deformation cause differences in impedance between flowing and resting blood. Through theoretical calculation and experimental measurements, we studied the effects of these factors on blood impedance. The size and shape of the erythrocyte and the conductivity of the interior medium of the erythrocyte change when the osmotic pressure of plasma is changed. From experimental results, we obtained the following: when the size of the erythrocyte becomes larger than the normal size due to the osmotic pressure change, the beta dispersion frequency decreases and the intra- and extracellular fluid resistance increase. These experimental results corroborate that the change of tissue impedance like muscle impedance during hemodialysis is caused by the change of the fluid distribution and the change of ionic concentration of the electrolyte in tissues during hemodialysis. Also, we could estimate the relative change value of the intra- and extracellular fluid volume by the impedance method, if there were no ionic concentration change in the electrolyte. It would be very difficult to estimate the absolute change value of them because a shadow effect due to the cells depends greatly upon the shape and size of the cells and the cell concentration.