G. Notas

Comparison of Ranson, APACHE II and APACHE III Scoring Systems in Acute Pancreatitis

Introduction Acute pancreatitis runs an unpredictable course. The early prediction of the severity of an acute attack has important implications for management and timely intervention. Aim To assess the prognostic accuracy of Ranson and APACHE II and III scoring systems in predicting the severity of acute pancreatitis. Methods One hundred fifty-three patients with acute pancreatitis (67.3% gallstone-related, 9.2% alcoholic, 17% idiopathic, and 6.5% of miscellaneous causes) were studied prospectively. Data conforming to the scoring systems were recorded 24 (the APACHE scores) and 48 hours (the Ranson score) after admission. Analysis was performed by using receiver operating characteristic curves (ROC), area under a curve (AUC), and by comparing likelihood ratios of positive test (LRPT). Results One hundred nineteen cases of pancreatitis were classified as mild, and 34 were classified as severe. The mortality rate was 3.2%. All three scores correlated with length of stay and disease severity. AUC for Ranson was found to be significantly larger than AUC for APACHE II and APACHE III score (0.817, cut-off ≥3; 0.618, cut-off, ≥10; and 0,676, cut-off ≥42 respectively). The Ranson score achieved the highest sensitivity and the lowest false-negative rate, but the positive and negative predictive values and LRPT were of similar extent for all three scores. Conclusion The APACHE III offers little, if any, advantage over the APACHE II score. Ranson criteria proved to be as powerful a prognostic model as the more complicated APACHE II and III scoring systems, but with the disadvantage of a 24-hour delay.

A comparison of Child-Pugh, APACHE II and APACHE III scoring systems in predicting hospital mortality of patients with liver cirrhosis

BackgroundThe aim of this study was to assess the prognostic accuracy of Child-Pugh and APACHE II and III scoring systems in predicting short-term, hospital mortality of patients with liver cirrhosis.Methods200 admissions of 147 cirrhotic patients (44% viral-associated liver cirrhosis, 33% alcoholic, 18.5% cryptogenic, 4.5% both viral and alcoholic) were studied prospectively. Clinical and laboratory data conforming to the Child-Pugh, APACHE II and III scores were recorded on day 1 for all patients. Discrimination was evaluated using receiver operating characteristic (ROC) curves and area under a ROC curve (AUC). Calibration was estimated using the Hosmer-Lemeshow goodness-of-fit test.ResultsOverall mortality was 11.5%. The mean Child-Pugh, APACHE II and III scores for survivors were found to be significantly lower than those of nonsurvivors. Discrimination was excellent for Child-Pugh (ROC AUC: 0.859) and APACHE III (ROC AUC: 0.816) scores, and acceptable for APACHE II score (ROC AUC: 0.759). Although the Hosmer-Lemeshow statistic revealed adequate goodness-of-fit for Child-Pugh score (P = 0.192), this was not the case for APACHE II and III scores (P = 0.004 and 0.003 respectively)ConclusionOur results indicate that, of the three models, Child-Pugh score had the least statistically significant discrepancy between predicted and observed mortality across the strata of increasing predicting mortality. This supports the hypothesis that APACHE scores do not work accurately outside ICU settings.

RT-PCR and immunocytochemistry studies support the presence of somatostatin, cortistatin and somatostatin receptor subtypes in rat Kupffer cells

The present study investigated the presence of somatostatin receptor subtypes (ssts) and the endogenous peptides somatostatin and cortistatin in rat Kupffer cells, since modulation of these cells by somatostatin may be important for the beneficial effect of somatostatin analogues in a selected group of hepatocellular carcinoma patients. Kupffer cells were isolated from rat liver in agreement with national and EU guidelines. RT-PCR was employed to assess the expression of somatostatin, cortistatin and ssts in Kupffer cells. Western blot analysis and immunocytochemistry were employed to assess the expression and the localization of the receptors, respectively. Quiescent Kupffer cells were found to express sst(1-4) mRNA, while immunocytochemical studies supported the presence of only the sst(3) and sst(4) receptors, which were found to be internalized. However, sst1 and sst(2A) receptors were detected by western blotting. RT-PCR and RIA measurements support the presence of both somatostatin and cortistatin. Stimulation of the cells with LPS activated the expression of the sst(2), sst(3) and sst(4) receptors. The present data provide evidence to support the presence of ssts and the endogenous neuropeptides somatostatin and CST in rat Kupffer cells. Both peptides may act in an autocrine manner to regulate sst receptor distribution. Studies are in progress in order to further characterize the role of ssts in Kupffer cells and in hepatic therapeutics.

Differential effects of somatostatin on tnf receptors and apoptosis in hepatocellular carcinoma cell lines

Introduction Somatostatin has antitumour activity in animal models of hepatocellular carcinoma. In human studies both favourable and unfavourable results have been reported. A 40% fraction of treated patients are benefited. Since its action may be through apoptosis, the authors tested the hypothesis that somatostatin may act differently in different hepatocellular cell lines. Methods The somatostatin synthetic analogue octreotide was tested for its effect on the expression of TNF receptors (RT-PCR and Western blot) and the TNFα-induced apoptosis (NF-kB nuclear translocation, P65 phosphorylation, DNA fragmentation and TUNEL cytochemistry) The HepG2 human hepatoblastoma cells and the Hep3B human hepatocellular carcinoma cells were used. Results TNFR1 but no TNFR2 receptor was constitutively expressed in both cell lines. Octreotide caused an early (within 1 h) reduction of both mRNA and TNFR1 protein in HepG2 and a late (at 6 and 12 h) reduction in Hep3B cells. TNFα induced NF-kB nuclear translocation, stronger in Hep3B which was not influenced by octreotide. Octreotide significantly increased P65Ser536 and P65Ser468 phosphorylation, more pronounced in Hep3B cells. Octreotide significantly decreased TNFα-induced Ser536 phosphorylation only in HepG2 cells. Octreotide or TNFα alone had no effect on apoptosis in HepG2 cells but TNFα greatly increased apoptosis in Hep3B cells. Co-incubation with Octreotide and TNFα increased apoptosis in HepG2 cells and decreased TNFα induced apoptosis in Hep3B cells, Conclusion (1) The differential effect of somatostatin on apoptosis may be due to its different effect on TNFR1 expression in the two cell lines. (2) Somatostatin may have no direct effect on apoptosis but it may indirectly act through TNF induced apoptosis. (3) This differential effect may in part explain the conflicting results of human HCC studies.

Differential effect of somatostatinergic signaling on collagen type i production and the proliferation of cytokine primed rat hepatic stellate cells

Introduction Somatostatin may influence hepatic fibrosis with mediators produced by Kupffer cells. The aim of this study is to investigate the role of somatostatinergic and cytokine signalling in hepatic stellate cells (HSC), the effector cells of hepatic fibrosis. Methods The production of a1-procollagen (a1-PROC) by rat HSCs treated with TNFα (100 ng/ml), TGFβ1 (5 ng/ml) or PDGF (32 ng/ml) and their cellular proliferation with or without octreotide was investigated. a1-PROC and aSMA were analysed by Western blotting and cellular proliferation was assayed by MTT. The role of the phosphotyrosine (PTP) and phosphoserine-phosphothreonine (STP) phosphatases on somatostatin signalling, was investigated by using the PTP inhibitor sodium orthovanadate (0.1 μM) and the STP inhibitor okadaic acid (0.1 μM). Results TGFβ1 and PDGF enhanced, whereas TNFα inhibited the expression of a1-PROC. Octreotide dose dependently inhibited the expression of a1-PROC in cells treated with TGFβ1, PDGF and increased the production of a1-PROC in TNF treated cells. Sodium orthovanadate significantly augmented the inhibition of a1-PROC production caused by octreotide only in TGFβ1 or PDGF treated cells. Okadaic acid uniformly inhibited the expression of a1-PROC. The expression of aSMA remained constant in all experiments. HSC proliferation increased by TGFβ1 and PDGF and was inhibited by TNFα Octreotide potentiated the effect of TGFβ1 and PDGF, and reversed TNFα inhibition. Orthovanadate and okadaic acid did not have any effect on the proliferation of cells. However, okadaic acid profoundly inhibited HSC proliferation when combined with octreotide, in TGFβ1 and PDGF treated cells. Conclusion Somatostatin differentially influences HSC a1 procollagen production according to cytokine microenvironment and this effect is modulated by tyrosine and threonin phosphatases. Proliferation of HSCs is similarly influenced by Somatostatin by a phosphatase dependent mechanism.

Somatostatin effect on TNFA receptors in Kupffer cells

Introduction Somatostatin is a mediator of immune functions and has been used as an antineoplastic agent in animal models and human neoplasias. We have demonstrated that octreotide-a somatostatin analogue- inhibits only LPS-induced secretion of pro-inflammatory cytokines including tumour necrosis factor (TNF) α by Kupffer cells.1 2 We therefore tested the hypothesis that somatostatin modulates the expression of TNFα receptors and apoptosis of Kupffer cells. Methods Rat Kupffer cells were isolated by centrifugal elutriation. TNFR1 and TNFR2 expression was studied by RT-PCR, quantitative PCR, Western Blot and immunofluorescence before and after octreotide pre-incubation. Apoptosis was assessed by quantitative measurement of cytoplasmic histone-associated DNA fragments (Roche). TNFα mRNa expression was assessed by a semiquantitative PCR and TNFα was measured in cell supernatants by ELISA. Results TNFR1 and TNFR2 m RNA are constitutively expressed in Kupffer cells. Octreotide incubation increased both receptors expression with a peak at 6 h and return to basal levels at 24 h. TNFR1 was mostly influenced. However only TNFR2 protein increase was identified by Western blot, while a band at 90 kD was present instead of a band at 55 kD as expected for TNFR1. TNFa mRNA expression and protein secretion was not influenced by octreotide at 24 h. However a significant inhibition was observed at 48 h. TNF had no effect on Kupffer cell apoptosis while octreotide significantly increased their apoptosis and this effect was not influenced by co-incubation with TNFα. Conclusion TNFR1 and TNFR2 are constitutively expressed in Kupffer cells and their expression is strongly increased by somatostatin. Moreover somatostatin increases Kupffer cell apoptosis. These findings may in part explain the antineoplasmatic effect of somatostatin.