Analysis of microvascular density in early gastric carcinoma using magnifying endoscopy with narrow-band imaging.

BACKGROUND AND STUDY AIMS: Intramucosal vascular density differs between differentiated and undifferentiated type gastric carcinomas. This study aimed to evaluate the microvascular density characteristics of these two types of carcinoma using magnifying endoscopy with narrow-band imaging (ME-NBI). PATIENTS AND METHODS: In total, 42 differentiated and 10 undifferentiated types were evaluated. The microvessels observed using ME-NBI were extracted from stored still images and the microvascular density in the two carcinoma types was analyzed. Histological vascular density in resected specimens was also evaluated using CD34 immunostaining. RESULTS: There were significant differences between the microvascular density in the differentiated and undifferentiated types of carcinoma (10.02 ±â€Š4.72 % vs 4.02 ±â€Š0.40 %; P < 0.001) using ME-NBI. Vascular density assessed histologically also differed significantly between differentiated and undifferentiated types in both the whole mucosal (5.81 ±â€Š3.17 % vs 3.25 ±â€Š1.21 %) and the superficial mucosal layers (0 - 100 µm) (6.38 ±â€Š3.73 % vs 3.66 ±â€Š1.46 %). However, the vascular density in the surrounding non-carcinomatous mucosa assessed using ME-NBI and histologically, was significantly lower in the differentiated than in the undifferentiated types (P < 0.001). There was good agreement between ME-NBI and histologically assessed microvascular density in both the whole (r = 0.740; P < 0.001) and superficial mucosal layers (r = 0.764; P < 0.001). White opaque substance (WOS) was seen in eight patients who had the differentiated type carcinoma. In almost all cases with WOS, the appearance of the carcinoma was discolored. CONCLUSIONS: There was a close relationship between ME-NBI assessed microvascular density and histologically assessed vascular density in the mucosal layer. Microvascular density differed significantly between the differentiated and undifferentiated types of carcinoma assessed using ME-NBI.

Factors associated with an absence of effect of an education program for improving knowledge of schizophrenia.

Schizophrenia can develop in junior and senior high school students. Correct identification of schizophrenia symptoms is an important factor in subsequent healthcare. The present study conducted a multifaceted evaluation of factors associated with an absence of effect of an education program for improving knowledge of schizophrenia among parents of Japanese junior and senior high school students. Regarding discrimination of prodromal symptoms, the factors associated with an absence of effect of the education program were graduate school education, family income >110 000 USD, proximity to a person with schizophrenia, employment as a professional, and participation in welfare activities for people with mental illness. Regarding discrimination of schizophrenia, the factors associated with an absence of effect were a family income of 53 000 to 110 000 USD (P<0.05), and employment in production/labor service (P<0.05).

Syntheses, structures, and antimicrobial activities of remarkably light-stable and water-soluble silver complexes with amino acid derivatives, silver(I) N-acetylmethioninates.

Reaction of L- and DL-N-acetylmethionine (Hacmet) and Ag(2)O in water at ambient temperature afforded the remarkably light-stable silver complexes {[Ag(L-acmet)]}(n) (1) and {[Ag(2)(D-acmet)(L-acmet)]}(n) (2), respectively. The color of the solids and aqueous solutions of 1 and 2 did not change for more than 1 month under air without any shields. The light stability of these two silver(I) complexes is much higher than that of silver(I) methioninate {[Ag(2)(D-met)(L-met)]}(n) (3) (Hmet = methionine), silver(I) S-methyl-L-cysteinate {[Ag(L-mecys)]}(n) (4), and silver(I) L-cysteinate {[Ag(L-Hcys)]}(n) (5). X-ray crystallography of 1 obtained by vapor diffusion revealed that ladder-like coordination polymers with two O- and two S-donor atoms were formed. The acetyl group of acmet(-) prevents chelate formation of the ligand to the metal center, which is frequently observed in amino acid metal complexes, but allows for formation of hydrogen bonds between the ligands in the crystals of 1. These two silver(I) N-acetylmethioninates showed a wide spectrum of effective antimicrobial activities against gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and yeasts (Candida albicans and Saccharomyces cerevisiae), the effectiveness of which was comparable to that of water-soluble Ag-O bonding complexes.

Encapsulation of anion/cation in the central cavity of tetrameric polyoxometalate, composed of four trititanium(IV)-substituted α-Dawson subunits, initiated by protonation/deprotonation of the bridging oxygen atoms on the intramolecular surface.

Preparation and structural characterization of a novel polyoxometalate (POM), [(P(2)W(15)Ti(3)O(60.5))(4)(NH(4))](35-) 1, i.e., an encapsulated NH(4)(+) cation species in the central cavity of a tetramer (called the Dawson tetramer) constituted by trititanium(IV)-substituted α-Dawson POM substructure, are described. POM 1 was synthesized by several different methods and unequivocally characterized by complete elemental analysis, thermogravimetric and differential thermal analysis (TG/DTA), FTIR spectroscopy, solution ((15)N{(1)H}, (31)P, (183)W) NMR spectroscopy, and X-ray crystallography. First, POM 1 was synthesized by a reaction of NH(4)Cl in aqueous solution with a precursor, which was derived by thermal treatment of a monomeric triperoxotitanium(IV)-substituted Dawson POM, [α-1,2,3-P(2)W(15)(TiO(2))(3)O(56)(OH)(3)](9-) 2, for 3 h in an electric furnace at 200 °C. The encapsulated NH(4)(+) cation in 1 was confirmed by (15)N{(1)H} NMR measurement and X-ray crystallography. As another synthesis of 1, a direct exchange of the Cl(-) anion encapsulated in [{α-1,2,3-P(2)W(15)Ti(3)O(57.5)(OH)(3)}(4)Cl](25-) 3 with the NH(4)(+) cation was attained by neutralizing an aqueous solution containing 3 with the addition of aqueous NH(3) (the initial pH of ca. 2-2.5 was changed to 6.4), followed by adding NH(4)Cl. It has been clarified that the conditions as to whether the anion or the cation is encapsulated in the central cavity of the Dawson tetramer were significantly related to the protonation/deprotonation of the bridging oxygen atoms on the intramolecular surface, Ti-O-Ti/Ti-OH-Ti sites constituting the Dawson subunits.

Intercluster compound between a tetrakis{triphenylphosphinegold(I)}oxonium cation and a keggin polyoxometalate (POM): formation during the course of carboxylate elimination of a monomeric triphenylphosphinegold(I) carboxylate in the presence of POMs.

The preparation and structural characterization of a novel intercluster compound, [{Au(PPh(3))}(4)(µ(4)-O)](3)[α-PW(12)O(40)](2)·4EtOH (1), constructed between a tetrakis{triphenylphosphinegold(I)}oxonium cation and a saturated α-Keggin polyoxometalate (POM) are described. The tetragold(I) cluster oxonium cation was formed during the course of carboxylate elimination of a monomeric phosphinegold(I) carboxylate complex, i.e., [Au((R,S)-pyrrld)(PPh(3))] [(R,S)-Hpyrrld = (R,S)-2-pyrrolidone-5-carboxylic acid], in the presence of the free acid form of a Keggin POM, H(3)[α-PW(12)O(40)]·7H(2)O. The liquid-liquid diffusion between the upper water/EtOH phase containing the Keggin POM and the lower CH(2)Cl(2) phase containing the monomeric gold(I) complex gave a pure crystalline sample of 1 in good yield (42.1%, 0.242 g scale). Complex 1 was formed by ionic interaction between the tetragold(I) cluster cation and the Keggin POM anion. As a matter of fact, the POM anion in 1 can be exchanged with the BF(4)(-) anion using an anion-exchange resin (Amberlyst A-27) in BF(4)(-) form. By using other Keggin POMs, such as H(4)[α-SiW(12)O(40)]·10H(2)O and H(3)[α-PMo(12)O(40)]·14H(2)O, the same tetragold(I) cluster cation was also formed, i.e., in the forms of [{Au(PPh(3))}(4)(µ(4)-O)](2)[α-SiW(12)O(40)]·2H(2)O (2) and [{Au(PPh(3))}(4)(µ(4)-O)](3)[α-PMo(12)O(40)](2)·3EtOH (3). Compounds 1-3, as dimethyl sulfoxide-soluble, EtOH- and Et(2)O-insoluble dark-yellowish white solids, were characterized by complete elemental analysis, thermogravimetric and differential thermal analyses, Fourier transform IR, X-ray crystallography, and solid-state (CPMAS (31)P and (29)Si) and solution ((31)P{(1)H} and (1)H) NMR spectroscopy. The molecular structures of 1 and 2 were successfully determined. The tetragold(I) cluster cation was composed of four PPh(3)Au(I) units bridged by a central µ(4)-oxygen atom in the geometry of a trigonal pyramid or distorted tetrahedron.

Synthesis and structure of dinuclear hafnium(IV) and zirconium(IV) complexes sandwiched between 2 mono-lacunary alpha-Keggin polyoxometalates.

The synthesis and characterization of two dinuclear HfIV and ZrIV complexes sandwiched between 2 mono-lacunary alpha-Keggin polyoxometalates (POMs), i.e., (Et2NH2)8[{alpha-PW11O39Hf(micro-OH)(H2O)}2].7H2O (Et2NH(2)-1) and (Et2NH2)8[{alpha-PW11O39Zr(micro-OH)(H2O)}2].7H2O (Et2NH(2)-2), are described. [Note: the moieties of their polyoxoanions are abbreviated simply as and , respectively.] A pair of HfIV- and ZrIV-containing POMs belonging to the same family were herein isolated as diethylammonium salts and were unambiguously characterized by complete elemental analysis, including sodium and oxygen analyses, TG/DTA, FT-IR, single-crystal X-ray structure analysis and solution (31P and 183W) NMR spectroscopy. Polyoxoanions 1 and 2 were isostructural with each other. The central [M2(micro-OH)2(H2O)2]6+ (M=Hf, Zr) cation unit was composed of 2 edge-sharing polyhedral M units, which were linked through 2 micro-OH groups and contained 1 water molecule coordinated to each metal center. Since the mono-lacunary Keggin POM acts as an oxygen-donor quadridentate ligand, the Hf and Zr centers are 7-coordinate. It should be noted that the present 2 Keggin 2:2-type compounds, Et2NH(2)-1 and Et2NH(2)-2, undergo a reversible conversion to Keggin 1:2-type complexes [M(alpha-PW11O39)2]10-, respectively, in solution under appropriate conditions. The synthesis of Et2NH(2)-1 and Et2NH(2)-2 is based on such an interconversion. The Zr compound Et2NH(2)-2 was rigorously compared with the 3 Zr POMs (OK-1-OK-3), recently reported by Kholdeeva's group: their POMs in a different protonation-state did not contain any coordinating water molecules.

Sandwich-type Hf(IV) and Zr(IV)complexes composed of tri-lacunary Keggin polyoxometalates: structure of [M(3)(mu-OH)(3)(A-alpha-PW(9)O(34))(2)](9-) (M = Hf and Zr).

The syntheses and structures of hafnium(iv) and zirconium(iv) complexes with Keggin tri-lacunary polyoxometalates (POM) [A-alpha-PW(9)O(34)](9-), i.e., (Et(2)NH(2))(7)H(2)[Hf(3)(micro-OH)(3)(A-alpha-PW(9)O(34))(2)].11H(2)O (Et(2)NH(2)-) and (Et(2)NH(2))(7)H(2)[Zr(3)(micro-OH)(3)(A-alpha-PW(9)O(34))(2)].12H(2)O (Et(2)NH(2)-) are described. The two POMs, Et(2)NH(2)- and Et(2)NH(2)-, were obtained as analytically pure, colorless crystals in 30.0 and 40.9% yields, respectively, by 1:3-molar ratio reactions of [A-PW(9)O(34)](9-) with Hf(SO(4))(2) and Zr(SO(4))(2), respectively, in aqueous solutions at 80-90 degrees C, followed by refluxing. The two POMs were characterized by elemental analysis, TG/DTA, FTIR, solution ((31)P and (183)W) NMR and X-ray crystallography. X-Ray structure analysis of Et(2)NH(2)- revealed that in polyoxoanion , the three 6-coordinate prismatic Hf(IV) ions linked with the three bridging OH groups, i.e., the [Hf(3)(micro-OH)(3)](9+) cluster cation was sandwiched between two alpha-Keggin tri-lacunary POMs. The molecular structure of was isostructural with that of polyoxoanion in Et(2)NH(2)-. X-Ray crystallography also showed that the crude crystals of Et(2)NH(2)- and Et(2)NH(2)-, which were first formed without refluxing, contained the two isomeric crystalline species in a 7:3 ratio with the alpha,alpha- and alpha,beta-junctions of the two Keggin tri-lacunary POMs. These facts were also confirmed by (31)P NMR in D(2)O. By refluxing the solutions containing the crude crystals, only the alpha,alpha-isomers of Et(2)NH(2)- and Et(2)NH(2)- were finally isolated.

Syntheses, molecular structures and pH-dependent monomer-dimer equilibria of Dawson alpha2-monotitanium(IV)-substituted polyoxometalates.

The preparation and structural characterization of a novel, solid Brønsted acid based on Dawson alpha2-monotitanium(IV)-substituted polyoxometalates (POMs) are described. The free-acid form of the POM, i.e., 13H+-heteropolyacid with the formula H13[1b].55H2O DH-1 (1b = [(alpha2-P2W17TiO61)(alpha2-P2W17TiO61H)(mu-O)]13-), was prepared by passing an aqueous solution containing a potassium salt precursor, K14[1a].17H2O DK-1 (1a=[(alpha2-P2W17TiO61)2(mu-O)]14-), through a cation-exchange resin column. Compound DK-1 was obtained by a stoichiometric reaction of mono-lacunary Dawson POM [alpha2-P2W17O61]10- with Ti(SO4)2 in an aqueous solution. [Note: the abbreviations D, M, K and H stand for dimer, monomer, potassium salt and free-acid form, respectively.] Compounds DK-1 and DH-1 were characterized by using complete elemental analysis, thermogravimetric (TG) and differential thermal analysis (DTA), FTIR, solution (31P and 183W) NMR spectroscopy, pH-varied 31P NMR spectroscopy, solid-state 31P CPMAS NMR, X-ray crystallography and acidity measurements in an organic solvent with a Hammett indicator for DH-1. The monomeric form, K8[alpha2-P2W17TiO62]. 18H2O MK-1, was derived from DK-1. The molecular structure of 1b was composed of a dimer connected through one Ti-O-Ti bond between two alpha2-mono-Ti(IV)-substituted Dawson POM subunits. BVS (bond valence sum) calculation showed that one oxygen atom (O(60A)) in one of the two Dawson subunits was protonated, therefore the two subunits were unequivalent. On the other hand, the molecular structure of la was a Ti-O-Ti bonding dimer of two equivalent Dawson subunits. The pH-varied 31P NMR spectra of DK-1 and DH-1 in aqueous solutions showed that the monomer at pH 7.0, the dimer at pH 1.0-3.0, and the mono-protonated species of the dimer at pH 0.5 were the predominant species in the solutions. The Hammett acidity constant (H0) of DH-1 in CH3CN (-2.87) was estimated to be almost the same as that of homo-Dawson heteropolyacid H6[P2W18O62].17H2O(-2.77).

Right ventricular involvement in a patient with isolated noncompaction of the ventricular myocardium.

Isolated noncompaction of the ventricular myocardium (INVM) is an unclassified cardiomyopathy and is thought to be due to arrest of myocardial morphogenesis. Although right ventricular involvement is not uncommon, the correct diagnosis is often difficult by echocardiography. In this report, we describe a patient with INVM in whom magnetic resonance imaging was useful to detect right ventricular morphological and functional abnormalities.

[Examination of the shapes of the end of the common bile ducts after endoscopic sphincterotomy (EST)].

At the treatment of bile duct stones using EST, we sometimes experienced the difficult cases, even if the stones were small. For these cases, we tried endoscopic papillary balloon dilatation, and as a result knoches were made. And after the disappearance of the knoch, we could easily remove the stone. So we prospectively examined how often and at what cases, the knoches were made immediately after EST. As a result, knoches were present for 25% (13/52) of patients even after EST. Narrow distal segments of knoch-present patients were longer than that of knoch-absent patients, significantly. As a result, some function of sphincter was shown to be remained for a quarter of patients even after EST, and for these cases, the treatment of the stones may be difficult.

[Two cases of patients with hepatocellular carcinoma (HCC) that developed in cryptogenic cirrhosis suggestive of nonalcoholic steatohepatitis (NASH) as background liver disease after clinical courses of 26 years].

We report two cases of patients with hepatocellular carcinoma (HCC) that developed in cryptogenic cirrhosis suggestive of nonalcoholic steatohepatitis (NASH) as background liver disease. Case 1 was a 68-year-old woman, and case 2 was a 46-year-old man. They were admitted to our department for evaluation and treatment of HCC. The causes of the underlying liver disease were not determined from blood tests. However, histological analysis of non-tumor tissues of the liver revealed cirrhosis with few fat droplets. Both patients had undergone liver biopsy 26 years before the treatment of HCC. Histological review of the biopsy specimens revealed NASH (case 1) and fatty liver (case 2), respectively. It was suggested that these cases progressed from NASH and fatty liver, respectively, to NASH-related cirrhosis (so called burned-out NASH), eventually, developing HCC. These findings suggest that substantial number of burned-out NASH cases may be included in those with cryptogenic cirrhosis. These two patients are indicative cases that may reveal the long-term natural course of fatty liver and NASH.

Perforation of the right ventricular free wall by an ICD lead in a patient with isolated noncompaction of the ventricular myocardium.

A 74-year-old patient with isolated noncompaction of the ventricular myocardium (INVM) underwent implantable cardioverter defibrillator (ICD) implantation because of sustained ventricular tachycardia. The post operative course was complicated by perforation of the right ventricular free wall by the ICD lead. The type of the active fixation transvenous ICD lead and the type of background cardiac disease are thought to be major risk factors for perforation.

Clinical and electrophysiological characteristics of binodal disease.

BACKGROUND: Although coexistence of atrioventricular conduction disturbances with sick sinus syndrome (SSS), so-called binodal disease (BND), is a frequently encountered disorder, its clinical significance and electrophysiological characteristics remain unknown. METHODS AND RESULTS: One hundred and seven patients with SSS were divided into BND (n=30) and N-BND groups (n=77). Sinus cycle length, sinus node recovery time (SRT), sino-atrial conduction time (SACT), the number of isolated sinus node electrograms, atrio-His (AH) interval, His-ventricular (HV) interval, intra-atrial conduction time (PA intervals) and QRS width were measured. In addition, the prevalence of bundle-branch block was obtained. The parameters of sino-atrial and intra-atrial conduction were significantly longer in the BND group: SRT (5,070+/-2,628 vs 3,122+/-1,856 ms, p<0.05), SACT (115+/-30 vs 87+/-21 ms, p<0.05), PA intervals (56+/-13 vs 41+/-8 ms, p<0.05). The BND group was more likely to have atrial fibrillation than the N-BND group (83.3% vs 53.2%, p<0.01). HV interval, QRS width and the prevalence of associated bundle-branch block did not differ between the 2 groups. CONCLUSION: BND patients not only had sino-atrial and atrioventricular node dysfunction, but also widespread atrial conduction disturbances. Thus, in the clinical setting BND should be categorized as severe SSS.

Inosine alleviates rat caerulein pancreatitis and pancreatitis-associated lung injury.

BACKGROUND: Recent studies have shown that inosine, a purine nucleoside produced during the breakdown of adenosine, has immunomodulatory and anti-inflammatory properties. The aim of this study was to examine the effects of inosine on the course of acute pancreatitis. METHODS: Edematous pancreatitis was induced by the intraperitoneal injection of caerulein (50 micro g/kg), seven times, at 1-h intervals, in male Wistar rats (caerulein pancreatitis). Inosine (100 mg/kg) was administered 30 min before or 1 h after the first injection of caerulein. The effects of inosine on the severity of pancreatitis were assessed by serum amylase, pancreatic edema (wet/dry ratio), myeloperoxidase activity, cytokine-induced neutrophil chemoattractant-1 concentrations, and histological changes. RESULTS: Prophylactic administration of inosine significantly decreased the elevation of serum amylase, myeloperoxidase activity, and cytokine-induced neutrophil chemoattractant-1 concentrations in the pancreas and the lung. Inosine did not significantly affect edema formation. Histologically, vacuole formation in pancreatic acinar cells, infiltration of inflammatory cells in the pancreas and the lung, and alveolar wall thickening in the lung were reduced. Inosine improved the histological findings and reduced myeloperoxidase activity even if it was administered 1 h after the first injection of caerulein. CONCLUSIONS: Inosine reduced the severity of acute pancreatitis, suggesting a possible application of this compound in the treatment of acute pancreatitis.

Tetrameric, trititanium(IV)-substituted polyoxotungstates with an alpha-Dawson substructure as soluble metal-oxide analogues: molecular structure of the giant "tetrapod" [(alpha-1,2,3-P2W15Ti3O62)4[mu3-Ti(OH)3]4Cl]45-.

The preparation and structural characterization of the novel polyoxoanion [(alpha-1,2,3-P(2)W(15)Ti(3)O(62))(4)[mu(3)-Ti(OH)(3)](4)Cl](45-) (1 a; abbreviated to [TiO(6)](16); FW approximately 16 000) which consists of four tri-Ti(IV)-1,2,3-substituted alpha-Dawson substructures, four Ti(OH)(3) bridging groups, and one encapsulated Cl(-) ion, are described. A water-soluble, all-inorganic composition compound of the tetrameric Ti-O-Ti-bridged anhydride form, Na(x)H(45-x)[1 a].y H(2)O (1; x=16-19, y=60-70), which was afforded by the reaction of the tri-lacunary Dawson polyoxotungstate Na(12)[B-alpha-P(2)W(15)O(56)].19 H(2)O with an excess of TiCl(4) in aqueous solution, was obtained as analytically pure, homogeneous colorless crystals. Single-crystal X-ray structure analysis revealed that 1 a was an inorganic, giant "tetrapod"-shaped molecule (inscribed to a sphere with a diameter of approximately 32 A) with approximately T(d) symmetry, in which the 16 edge- and/or corner-shared TiO(6) octahedra were contained. This number of TiO(6) octahedra was larger than that found in other titanium(IV)-substituted polyoxotungstates. Complex 1 was characterized by complete elemental analysis, TG/DTA, FTIR, UV/Vis absorption, and solution ((31)P and (183)W) NMR spectroscopy. The longest wavelength band in the UV/Vis absorption spectra of 1 in water was attributed to the O-->Ti(IV) ligand-to-metal charge-transfer (LMCT) transition: the wavelength of the LMCT band increased linearly as the number of TiO(6) octahedra contained in the Keggin and Dawson polyoxoanions increased. The Ti(n) chromophores formed in the Keggin and Dawson polyoxotungstates are water-soluble analogues of solid TiO(2) or SrTiO(3) as light-semiconductors and photocatalysts.

Nuclear factor kappa B expression in peripheral blood mononuclear cells of patients with acute pancreatitis.

INTRODUCTION: Although many experimental studies have implicated the activation of nuclear factor kappa B (NF-kappaB) as a pivotal step in the pathobiology of acute pancreatitis, no clinical investigations have been reported. AIMTo investigate the expression of NF-kappaB and its characteristics in peripheral blood mononuclear cells (PBMCs) of patient with acute pancreatitis. METHODOLOGY: Forty-five patients were prospectively enrolled. The expression of NF-kappaB in PBMCs was measured in the patients by electrophoretic mobility shift assay at admission and 14 days after the onset of acute pancreatitis. Twelve healthy individuals were also included as control subjects. RESULTS: At admission, the PBMCs from patients with acute pancreatitis showed higher levels of NF-kappaB activities than did those from control subjects. In vitro, the lipopolysaccharide (LPS) treatment of the PBMCs from the control subjects and patients with mild pancreatitis induced further activation of the NF-kappaB. The response was significantly reduced in patients with severe pancreatitis. Patients who had persistently high NF-kappaB activities, a reduced response of NF-kappaB to LPS, and a low p50p65:p50p50 ratio after LPS stimulation at 14 days developed serious systemic complications in the later clinical course. CONCLUSIONS: An alteration of the characteristics of PBMCs occurs in the early phase of acute pancreatitis and may predispose patients to a higher risk of serious systemic complications.

Macrophage migration inhibitory factor is a critical mediator of severe acute pancreatitis.

BACKGROUND & AIMS: Macrophage migration inhibitory factor (MIF), originally described as an inhibitor of the random migration of macrophages, has been shown recently to be involved in the pathogenesis of several inflammatory diseases such as sepsis. The aim of this study was to clarify the role of MIF in acute pancreatitis (AP). METHODS: Hemorrhagic necrotizing pancreatitis and edematous pancreatitis were induced by the injection of taurocholic acid (TCA pancreatitis) and cerulein (cerulein pancreatitis), respectively, on male Wistar rats. MIF levels in ascitic fluids, serum, and the organs were determined. The effects of anti-MIF antibody were examined on the prognosis of rats with TCA pancreatitis and of female CD-1 mice with choline-deficient, ethionine-supplemented, diet-induced model of severe AP. In addition, serum MIF levels in AP patients and in healthy controls were measured. RESULTS: Serum and ascitic MIF levels in TCA pancreatitis were increased rapidly and decreased gradually thereafter. Ascitic MIF levels were also increased in cerulein pancreatitis, but to a lesser degree. MIF level was increased in the lung in TCA pancreatitis, but not in the pancreas and the liver. Prophylactic (1 hour before and immediately after induction) administration of anti-MIF antibody significantly improved the survival rate of rats with TCA pancreatitis. The survival rate of mice with severe AP was also improved significantly by the antibody treatment. Serum MIF levels were higher in severe AP patients than mild AP patients and healthy controls. CONCLUSIONS: These results suggest a role of MIF in the pathogenesis of severe AP.

Inhibition of p38 mitogen-activated protein kinase blocks activation of rat pancreatic stellate cells.

Activated pancreatic stellate cells (PSCs) have recently been implicated in the pathogenesis of pancreatic fibrosis and inflammation. However, the signal transduction pathways in PSCs remain largely unknown. We examined the role of p38 mitogen-activated protein (MAP) kinase in the activation of PSCs. PSCs were isolated from rat pancreas tissue and used in their culture-activated, myofibroblast-like phenotype. Activation of p38 MAP kinase was determined by Western blotting using anti-phosphospecific antibody. The effects of two p38 MAP kinase inhibitors, 4-(4-flurophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)imidazole (SB203580) and 4-(4-flurophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole (SB202190), on the parameters of PSC activation, including proliferation, expression of alpha-smooth muscle actin, alpha1(I) procollagen, and prolyl 4-hydroxylase (alpha) genes, and monocyte chemoattractant protein-1 production were evaluated. Interleukin-1beta and platelet-derived growth factor-BB activated p38 MAP kinase. Platelet-derived growth factor-induced PSC proliferation was inhibited by SB203580 and SB202190. These reagents decreased alpha-smooth muscle actin protein expression, and alpha1(I) procollagen and prolyl 4-hydroxylase (alpha) mRNA levels. Treatment with these p38 MAP kinase inhibitors also resulted in inhibition of monocyte chemoattractant protein-1 expression. In addition, SB203580 inhibited spontaneous activation of freshly isolated PSCs in culture on plastic. Thus, inhibition of p38 MAP kinase modulated profibrogenic and proinflammatory actions in PSCs, implying a potential application of p38 MAP kinase inhibitors for the treatment of pancreatic fibrosis and inflammation.

Establishment and characterization of a simian virus 40-immortalized rat pancreatic stellate cell line.

Activated pancreatic stellate cells (PSCs) have recently been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line would be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish an immortalized cell line of rat PSCs. PSCs were isolated from the pancreas of male Wistar rats, and the simian virus 40 T antigen was introduced to PSCs by retrovirus-mediated gene transfer. This procedure yielded an actively growing cell line, designated as SAM-K. This cell line has been passaged repeatedly for almost 2 years, and is thus likely immortalized. SAM-K cells retained morphological characteristics of primary PSCs, and expressed alpha-smooth muscle actin, glial fibrillary acidic protein, type I collagen, fibronectin, and prolyl hydroxylases. The level of p53 expression was very high in SAM-K cells. Proliferation of SAM-K cells was stimulated by serum and platelet-derived growth factor-BB. Interleukin-1beta (IL-1beta) activated nuclear factor-kappaB, activator protein-1, and three classes of mitogen-activated protein (MAP) kinases: extracellular signal-regulated kinase1/2, c-Jun N-terminal kinase, and p38 MAP kinase. IL-1beta induced expression of intercellular adhesion molecule-1 and monocyte chemoattractant protein-1, both of which were abolished in the presence of pyrrolidine dithiocarbamate, a specific inhibitor of nuclear factor-kappaB activation. IL-1beta-induced monocyte chemoattractant protein-1 was partially inhibited by specific inhibitors of MAP kinase kinase (U0126) and of p38 MAP kinase (SB203580) whereas intercellular adhesion molecule-1 expression was not altered by the inhibitors. Thus, SAM-K would be useful for in vitro studies of cell biology and signal transduction of PSCs.