Gene expression profiling of human liver transplants identifies an early transcriptional signature associated with initial poor graft function.

Liver ischemia-reperfusion injury occurring in orthotopic liver transplantation (OLT) may be responsible for early graft failure. Molecular mechanisms underlying initial poor graft function (IPGF) have been poorly documented in human. The purpose of this study was to identify the major transcriptional alterations occurring in human livers during OLT. Twenty-one RNA extracts derived from liver transplant biopsies taken after graft reperfusion were compared with 7 RNA derived from normal control livers. Three hundred seventy-one genes were significantly modulated and classified in molecular pathways relevant to liver metabolism, inflammatory response, cell proliferation and liver protection. Grafts were then subdivided into two groups based on their peak levels of serum aspartate amino transferase within 72 h after OLT (group 1, non-IPGF: 14 patients; group 2, IPGF: 7 patients). The two corresponding data sets were compared using a supervised prediction method. A new set of genes able to correctly classify 71% of the patients was defined. These genes were functionally associated with oxidative stress, inflammation and inhibition of cell proliferation. This study provides a comprehensive picture of the transcriptional events associated with human OLT and IPGF. We anticipate that such alterations provide a framework for the elucidation of the molecular mechanisms leading to IPGF.

Liver apoptosis following normothermic ischemia-reperfusion: in vivo evaluation of caspase activity by FLIVO assay in rats.

Normothermic liver ischemia-reperfusion (I-R) may induce hepatocellular apoptosis. Caspase activation is involved in the initiation and execution of apoptosis. The aim of this study was to determine in vivo caspase activity in normothermic liver I-R in rats. Segmental normothermic ischemia of the liver was induced for 120 minutes in rats. After intravenous injection of the green probe FLIVO, in vivo caspase-3- and -7-specific activity was determined using fluorescence microscopy, in either nonischemic or ischemic liver lobes at 3 and 6 hours after reperfusion. Liver apoptosis was assessed by the deoxynucleotide transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay. Fluorescence microscopy showed that in vivo caspase-3- and -7-specific activities were significantly increased (P< .005) in ischemic lobes at 3 and 6 hours of reperfusion, compared with nonischemic liver lobes. Quantitative analysis of apoptotic cells measured by the TUNEL method showed a significant increase among apoptotic cells in ischemic lobes at 3 and 6 hours after reperfusion (P< .005), compared with nonischemic liver lobes. In conclusion, 120-minute normothermic liver I-R resulted in increased caspase-3- and -7-specific activities and in liver cell apoptosis.

Liver HIF-1 alpha induction precedes apoptosis following normothermic ischemia-reperfusion in rats.

Apoptosis plays an important role in ischemia-reperfusion (I-R) injury during liver transplantation. The hypoxia-inducible factor alpha (HIF-1alpha) may trigger liver apoptosis following I-R through the induction of hypoxically regulated genes. The aim of this study was to evaluate the effect of normothermic liver I-R on HIF-1alpha expression and apoptosis in rats. Segmental normothermic ischemia of the liver was induced in rats for 120 minutes. Liver extracts from either ischemic or nonischemic lobes were prepared at 0, 1, 3, and 6 hours after reperfusion. Liver HIF-1alpha protein expression was examined by Western blot analysis. Liver apoptosis was quantified using terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick end labeling assay. Normothermic I-R resulted in a significant (P< .05) increase in liver HIF-1alpha protein levels 1 and 3 hours after reperfusion. Liver apoptosis was significantly (P< .005) increased at 3 and 6 hours after reperfusion. In conclusion, normothermic liver I-R leads to increased liver expression of HIF-1alpha and apoptosis.

Peritoneal implantation of cryopreserved encapsulated porcine hepatocytes in rats without immunosuppression: viability and function.

The bioartificial liver (BAL) represents a promising approach to cell transplantation without immunosuppression as a method to support patients with hepatic insufficiency. The aim of this study was to assess viability and function of cryopreserved encapsulated porcine hepatocytes implanted intraperitoneally in rats without immunosuppression. Isolated porcine hepatocytes were cryopreserved at -196 degrees C for 1 month. Four groups were created: group 1 (n=10), freshly encapsulated porcine hepatocytes cultured in albumin-free medium for 10 days; group 2 (n=10), freshly encapsulated porcine hepatocytes implanted in the rat peritoneum without immunosuppression for 1 month and cultured for 10 days after explantation; group 3 (n=10), cryopreserved encapsulated porcine hepatocytes cultured for 10 days; group 4 (n=10), cryopreserved encapsulated porcine hepatocytes implanted in the rat peritoneum without immunosuppression for 1 month and cultured for 10 days after explantation. We assessed urea and albumin production and hepatocyte viability. The hepatocytes of all groups retained the capacity to produce urea and albumin, although the albumin synthesis was significantly decreased among hepatocytes of group 4 (P< .01). Encapsulated cryopreserved porcine hepatocytes explanted from rat peritoneum after 1 month appeared morphologically viable; their ultrastructure was preserved. In conclusion, long-term cryopreservation of porcine hepatocytes resulted in retention of their biological activity and in significant viability when transplanted into the rat peritoneum without immunosuppression.

Inhibition of tumor necrosis factor alpha gene transcription by pentoxifylline reduces normothermic liver ischemia-reperfusion injury in rats.

Pentoxifylline (PTX) has been shown to protect the liver against normothermic ischemia-reperfusion (I-R) injury. The aims of this study were to investigate the action of PTX on tumor necrosis factor alpha (TNFalpha) gene transcription following normothermic liver I-R as well as to evaluate the resulting effects on liver function and survival. A segmental normothermic liver ischemia was induced for 90 minutes. Rats were divided into three groups: group 1, control, Ringer lactate administration; group 2, PTX treatment; group 3, sham-operated control rats. PTX (50 mg/kg) was injected intravenously 30 minutes before induction of ischemia and 30 minutes before reperfusion. The nonischemic liver lobes were resected at the end of ischemia. Survival rates were compared and serum activities of TNFalpha, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase were measured. Liver histology was assessed 6 hours after reperfusion. Liver TNFalpha mRNA was assessed by polymerase chain reaction amplification at different times after reperfusion. PTX treatment significantly decreased serum activities of TNFalpha and inhibited liver expression of TNFalpha mRNA. The extent of liver necrosis and serum levels of liver enzymes were significantly decreased by PTX treatment, resulting in a significant increase in 7-day survival compared with nontreated control rats. In conclusion, PTX inhibits liver TNFalpha gene transcription, decreases serum TNFalpha levels, and reduces liver injury following normothermic I-R.

Alterations in protein tyrosine kinase pathways in rat liver following normothermic ischemia-reperfusion.

The phosphoregulation of signal transduction pathways is a complex series of reactions that modulate the cellular response to ischemia-reperfusion (I-R). The aim of this study was to evaluate the effect of normothermic liver I-R on protein tyrosine phosphorylation, production of angiogenic growth factors, and activation of signal proteins in tyrosine kinase pathways. A segmental normothermic ischemia of the liver was induced in rats by occluding the blood vessels (including the bile duct) to the median and left lateral lobes for 120 minutes. Liver extracts from either ischemic or nonischemic lobes were prepared at 0, 1, 3, and 6 hours after reperfusion. Liver tyrosine phosphorylation of proteins was examined by Western blot analysis, whereas vascular endothelial growth factor (VEGF) mRNA was analyzed by Northern blot. In ischemic liver lobes, VEGF mRNA and total protein levels increased at 1 and 3 hours after reperfusion. Tyrosine phosphorylation of the VEGF receptor Flk-1 and the platelet-derived growth factor receptor (PDGF-R) was increased only at 1 hour after reperfusion, while c-Src tyrosine phosphorylation remained increased at 3 hours and remained up to 6 hours after reperfusion. In conclusion, 1-R led to alterations in protein tyrosine phosphorylation and increased expression of VEGF in rat liver.

Intestinal lactase-phlorizin hydrolase (LPH): the two catalytic sites; the role of the pancreas in pro-LPH maturation.

Brush border lactase-phlorizin hydrolase carries two catalytic sites. In the human enzyme lactase comprises Glu-1749, phlorizin hydrolase Glu-1273. The proteolytic processing of pro-lactase-phlorizin hydrolase by (rat) enterocytes stops two amino acid residues short of the N-terminus of 'mature' final, brush border lactase-phlorizin hydrolase. Only these two amino acid residues are removed by luminal pancreatic protease(s), probably trypsin.

Iatrogenic bladder stone formation on absorbable suture 3-years after radical prostatectomy.

Vesical calculi formation on absorbable sutures is rare. The case of a 68-year-old white man, who had formed a large bladder stone on absorbable suture 3 years after radical prostatectomy, is reported. Endoscopic lithotripsy of the bladder calculi was performed and the suture was removed.

Laparoscopic tumorectomy of renal carcinoma recurred in remaining kidney.

Nephron sparing surgery is the standard treatment for small, peripherally located renal cell carcinoma. To reduce the morbidity of nephron sparing surgery laparoscopy was proposed. A case of renal carcinoma recurred in the remaining kidney successfully treated by laparoscopic approach is reported.

Successful transarterial embolization of idiopathic renal arteriovenous fistula.

A case of idiopathic renal arteriovenous fistula in a 46-year-old woman presenting intensive right renal colic associated to massive hematuria is reported. The renal arteriography confirmed the diagnosis and embolization of the fistula was performed. The transarterial embolization was successful no recurrence is observed after one year follow-up.

[The biocompatibility of suture materials used in colon surgery. An experimental study in the rat]. / Biocompatibilità dei materiali di sutura in uso nella chirurgia del colon. Studio sperimentale nel ratto.

BACKGROUND: The aim of this experimental study was to compare the tissue behavior of biofragmentable anastomotic ring (BAR) with other synthetic materials used in colonic surgery. METHODS: Thirty-three rats were divided into four groups: group 1, sham-operated control animals without material implanted; group 2, with fragments of polypropylene monofilament implanted extraperitoneally in abdominal wound, between musculature and peritoneum; group 3, with metal clips implanted extraperitoneally in abdominal wound, between musculature and peritoneum and group 4, with fragments of biofragmentable anastomotic ring implanted extraperitoneally in abdominal wound, between musculature and peritoneum. Animals were sacrificed 30 days after the operation. Macroscopic and histological criteria were used to characterize the resistance of the wound and the tolerance of the host to the foreign material. RESULTS: The inflammatory cell reaction of host tissue was significantly greater in group 4 compared with other groups (p < 0.05). In three cases, in group 4, we observed the adhesion of implanted fragment to epiploa. The enumeration of giant cells and the degree of fibrotic reaction was similar in all groups with material implanted, but no significant difference between the groups was observed. Our findings showed the greater biocompatibility of polypropylene and metallic clips, compared to the biofragmentable ring anastomosis. The strong inflammatory reaction in the host tissue caused by biofragmentable anastomotic ring may explain partially clinical postoperative complications (anastomotic wound infection and/or dehiscence and/or stricture). CONCLUSIONS: In conclusion, the choice of suture materials should be based not only on the mechanical properties, but also on their biological interactions between host and suture materials and on the evaluation of their effective cost/benefit.

[Laparoscopic treatment of appendiceal peritonitis in adults]. / Traitement par voie coelioscopique des péritonites appendiculaires chez l'adulte.

UNLABELLED: The aim of this study was to evaluate the results of laparoscopic treatment of appendicular peritonitis. PATIENTS AND METHODS: From January 1991 to December 1994, 32 patients (16 men and 16 women with a mean age of 43 years) underwent emergency laparoscopy for a clinical diagnosis of localized or generalized appendicular peritonitis. All patients had double antibiotic therapy for at least 7 days. The laparoscopic appendectomy technique consisted of:insufflation to 12 mmHg, introduction of 3 trocars, first peritoneal lavage, coagulation of the mesoappendix, ligature of the base of the appendix, no drainage. RESULTS: There were 4 conversions (12.5%). Nine of the 28 cases treated completely by laparoscopy, presented generalized peritonitis and 19 presented localized peritonitis (including 8 abscesses). The operations were performed by 7 surgeons and the mean operating time was 86 minutes. There were no deaths. The postoperative morbidity was 10.7%. The mean duration of postoperative ileus was 2.8 days. The mean hospital stay was 6.8 days. Histological examination concluded on acute suppurative appendicitis 96.4% of cases. There were no bowel obstructions or incisional hernias with a mean followup of 28.5 months. CONCLUSIONS: The laparoscopic treatment of appendicular peritonitis is possible, simple and reproducible, effective, without any specific complications. The advantages of laparoscopic techniques over the traditional large incisions are the absence of parietal complications, the quality of exploration and peritoneal lavage, and improvement of postoperative comfort.

[Ischemia-reperfusion]. / Ischémie-reperfusion.

IMPORTANCE OF ISCHEMIA-REPERFUSION LESIONS: After transplantation, ischemia-reperfusion lesions are associated with an increased risk of acute rejection, late recovery of liver function, or chronic graft dysfunction. In all, about 20% of the grafts are lost. The importance of prevention is evident. HEME-OXYGENASE: It has been shown that heme-oxygenase, an anti-oxidant reducing apoptosis, reduces the extent of ischemia-reperfusion lesions after liver, heart, kidney or Langerhans islet transplantation. OTHER COMPOUNDS WITH INTERESTING PROPERTIES: Other compounds also have interesting properties for preventing ischemia-reperfusion lesions: a specific metallo-protease inhibitor, L-arginine, a selective agonist of the PGE1 receptor, estrogens, low-dose cyclosporine, and certain immunosuppressors (FTY 720, anti CD28, anti B7-1), and rPSGL-Ig ligand.

Induction of different types of cell death after normothermic liver ischemia-reperfusion.

Normothermic liver ischemia-reperfusion (I-R) may induce hepatocellular autophagy, apoptosis, and necrosis. The aim of this study was to investigate these three types of cell death in normothermic liver I-R in rats. A segmental normothermic ischemia of the liver was induced for 120 minutes. Liver autophagy was evaluated by transmission electron microscopy and LC3 (Light Chain 3) immunohistochemical studies. Liver apoptosis was assessed by FLIVO (FLuorescence in vIVO) and TUNEL (TdT-mediated dUTP nick end labeling) assays. Liver necrosis was determined by optical microscopic examination. Autophagy was increased in ischemic liver lobes at 6 hours after reperfusion, compared with nonischemic lobes. Fluorescence microscopy showed in situ caspase-3 and -7 specific activity to be increased in ischemic liver lobes after 6 hours of reperfusion, compared with nonischemic lobes. Quantitative analysis of apoptotic cells evaluated by the TUNEL method showed a clearly significant increase in ischemic liver lobes at 6 hours after reperfusion, compared with nonischemic lobes. Necrotic cell death was significantly increased in ischemic liver lobes at 6 hours after reperfusion, compared with nonischemic lobes (P < .005). In conclusion, 120 minutes normothermic liver I-R resulted in increased autophagic, apoptotic and necrotic cell death.

Cryopreserved porcine hepatocytes: expression and induction of cytochrome P450, isoform CYP2E1.

Cryopreservation of porcine hepatocytes for their use in bioartificial liver devices may result in reduced cytochrome P450 (CYP) enzyme activity. The aim of this study was to assess the effects of several CYP inducers on the isoform CYP2E1 protein expression in cryopreserved porcine hepatocytes. Isolated porcine hepatocytes were cryopreserved for 1 month, thawed, and cultured for 3 days. During medium culture, the hepatocytes were exposed to the following CYP inducers: dimethyl sulfoxide, rifampicin, phenobarbital, 3-methylcholanthrene, and dexamethasone. CYP2E1 protein expression was determined by immunoblotting. CYP2E1 protein levels were constantly detected in cryopreserved porcine hepatocytes. CYP inducers did not modify CYP2E1 protein levels. Long-term cryopreserved porcine hepatocytes preserved their capacity for CYP2E1 protein expression, although exposure of these hepatocytes to CYP inducers did not modify the CYP2E1 protein expression.