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J Domergue

Publications and source records attributed to J Domergue.

At least 55 records · Page 3Linked to original sources
PubMed1997

Long-term primary cultures of adult human hepatocytes.

In this work we have investigated a system of long-term primary cultures of adult human hepatocytes which, in contrast to those previously described, has the advantage of requiring neither the use of additive cells as in co-cultures, nor of matrix component preparations like Matrigel or collagen sandwich. This system has been used previously for long-term cultures of hepatocytes from young baboon, and some modifications have been introduced here to take into account the specificity of adult human hepatocytes. In this system, hepatocytes are plated at confluence on collagen-coated dishes and cultured in a serum-free medium consisting of Williams'E supplemented with hormones and growth factors. Proteins secreted specifically by the liver, including albumin, alpha-1 antitrypsin, plasminogen, fibrinogen, lipoproteins ApoA1 and ApoB100, have been quantified in the extracellular medium as a function of time, either by immunoblot or ELISA. In addition, the expression and inducibility of CYP proteins of the CYP1, CYP2 and CYP3 families in response to their prototypical inducers including 2,3,7,8-tetrachlorodibenzo(p)dioxin and rifampicin, have been evaluated by immunoblot analysis of microsomes or cell lysates. Moreover, the oxidative metabolism of cyclosporin A, a monoxygenase activity depending on CYP3A4, has been monitored directly on the cultured cells by HPLC analysis of extracellular medium. Our results show that, under these culture conditions, adult human hepatocytes retain these phenotypical characteristics for at least 35 days. This system meets the requirements for use as a model for screening CYP protein inducers.

Aged↗
PubMed1997

Lipid-mediated transfection of normal adult human hepatocytes in primary culture.

The aim of this work was to develop a procedure for the lipid-mediated transfection of DNA into normal adult human hepatocytes in culture. Cells were plated in a serum-free culture medium at various cell densities, on plastic or collagen-coated dishes, both in the absence and in the presence of epidermal growth factor (EGF). The cells were incubated for various periods of time with mixtures of DNA-lipofectin or DNA-3 beta[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-chol) liposomes, and the efficiency of transfection was assessed by measuring the activity of reporter genes, beta-galactosidase or chloramphenicol acetyl-transferase (CAT). For comparison, similar experiments were carried out with human cell lines including HepG2, Caco-2, and WRL68. The efficiency of transfection (in percentage of cells) was not significantly different after transfection with lipofectin or DC-chol and comprised between 0.04 and 1.7% (extreme values) for different cultures. The efficiency of transfection decreased as the age or density of the culture increased and increased in cultures treated with EGF. Direct measurement of the rate of DNA synthesis suggested that the efficiency of transfection was related to the number of cells entering the S phase. Under the same conditions, the efficiency of transfection was one to two orders of magnitude greater in the three cell lines. A plasmid harboring 660 bp of the 5'-flanking region of CYP1A1 (containing two xenobiotic enhancer elements) fused upstream of the promoter of thymidine kinase and the CAT reporter gene was constructed. When this plasmid was transfected in human hepatocytes, CAT activity was induced as expected. We conclude that normal adult human hepatocytes can be transfected with exogenous DNA and that the transfected construct is regulated in the manner expected from in vivo studies.

Adult↗
PubMed1997

Effect of cell density and epidermal growth factor on the inducible expression of CYP3A and CYP1A genes in human hepatocytes in primary culture.

The influence of cell density and epidermal growth factor (EGF) on the expression and inducibility of cytochrome P450 (CYP) genes of the CYP3A and CYP1A families in adult human hepatocytes in primary culture has been evaluated. Only when cultured at subconfluence and in the presence of EGF did hepatocytes exhibit a proliferative response, assessed by measuring DNA synthesis and cyclin A accumulation. In the absence of EGF, the accumulation of CYP3A4 and CYP1A2 messenger RNAs (mRNAs) in response to their respective inducers (rifampicin and dioxin) was dramatically decreased in subconfluent culture with respect to confluent cultures. The presence of EGF only slightly decreased the accumulation of these mRNAs in both confluent and subconfluent cultures. The accumulation of CYP2D6 and CYP2E1 proteins, which are constitutively expressed in confluent cultures, and the production of fibrinogen and apolipoprotein (Apo) B100 exhibited similar behavior, while nicotinamide adenine dinucleotide phosphate cytochrome c reductase activity was affected neither by cell density nor by EGF. In contrast, the accumulation of CYP1A1 mRNA in response to dioxin was similar in confluent and subconfluent cultures, irrespective of the presence of EGF. Interestingly, CYP3A7, a gene that is preferentially expressed in the fetal liver, was expressed constitutively neither in confluent nor in subconfluent cultures, irrespective of the presence of EGF. It is concluded that the loss of cell-cell contacts rather than the proliferative status of cells per se is responsible for the dramatic decrease in the expression of CYP genes, normally expressed in the adult human liver.

Adult↗
PubMed1997

Hepatitis C virus genotypes and quantitation of serum hepatitis C virus RNA in liver transplant recipients: relationship with severity of histological recurrence and implications in the pathogenesis of HCV infection.

The reasons for the wide variation of incidence and severity of recurrent hepatitis C after liver transplantation are not clear. We have studied liver transplant recipients to assess the impact of hepatitis C virus (HCV) genotype and HCV RNA quantification on HCV recurrence after transplantation. Twenty-two patients received transplants for HCV cirrhosis and were followed up with virological and histological assessments. Mean follow-up was 39 months. HCV genotype was determined with line probe assay (Inno-Lipa). HCV RNA quantity was determined in serum samples by use of polymerase chain reaction nested assay. HCV genotype 1 was detected in 13 patients and other genotypes in 9. Histological recurrence rates were 69% in patients with genotype 1 and 66% in patients with other genotypes. All cases of severe histological injury (chronic active hepatitis or cirrhosis) were observed in patients with genotype 1. HCV RNA quantity was significantly higher in patients with genotype 1 (mean, 2.023 x 10(3) copies/mL) than in patients with other genotypes (mean, 27,403 copies/mL). In conclusion, the severity of histological recurrence after liver transplantation for HCV disease was higher in patients infected by HCV genotype 1 than in those infected with other genotypes. The levels of viral replication were higher in patients with HCV genotype 1 than in those with other genotypes.

Adult↗
PubMed1997

Liver resection or transplantation for hepatocellular carcinoma? Retrospective analysis of 215 patients with cirrhosis.

BACKGROUND/AIMS: Currently, surgical treatment of hepatocellular carcinoma in patients with cirrhosis is not clearly defined. The objective of this study was, in patients with cirrhosis with hepatocellular carcinoma, to compare liver resection to transplantation assessed by patient survival and to determine whether the tumor recurrence might be influenced by prognostic factors. METHODS: We have gathered all the available data from six French Medical Universities, for 215 patients with cirrhosis with hepatocellular carcinoma surgically treated either by liver resection (102) or by transplantation (113). RESULTS: The overall 5-year survival rate was similar in the transplantation group and in the resection group (32% vs. 31%, p=0.7). However, the 5-year survival rate without recurrence was higher in the transplantation group than in the resection group (60% vs. 14%, p<0.001). Three independent prognostic factors influenced significantly the survival without recurrence: the surgical treatment by transplantation (p<0.001), the number of tumors (p<0.01) and the tumor size (p<0.001). With these factors we defined a prognostic index (Ip) which allowed assessment of the probability of survival without recurrence: Ip= (Xie. x 1.41)+(Nbr T. x 0.19)+(Size TV. x 0.16); Xie=surgical treatment (Xie=0 if transplantation, Xie=1 if resection), Nbr.T. and Size TV.=number of tumors and size of the most voluminous tumor, respectively, according to the histologic study. CONCLUSIONS: These results and this prognostic index are encouraging for liver transplantation as treatment of hepatocellular carcinoma in selected patients with cirrhosis.

Adult↗
PubMed1997

Results of liver transplantation in the treatment of metastatic neuroendocrine tumors. A 31-case French multicentric report.

OBJECTIVE: The purpose of this study was to assess the value and timing of orthotopic liver transplantation (OLT) in the treatment of metastatic neuroendocrine tumors (NET). SUMMARY BACKGROUND DATA: Liver metastasis from NET seems less invasive than other secondary tumors. This observation suggests that OLT may be indicated when other therapies become ineffective. However, the potential benefit of this highly aggressive procedure is difficult to assess due to the scarcity and heterogeneity of NET. METHODS: A retrospective multicentric study was carried out, including all cases of OLT for NET performed in France between 1989 and 1994. There were 15 cases of metastatic carcinoid tumor and 16 cases of islet cell carcinomas. Hormone-related symptoms were present in 16 cases (55%). Only 5 patients (16%) had no previous surgical or medical therapy before OLT. Median delay from diagnosis of liver metastasis and OLT was 19 months (range, 2 to 120). RESULTS: The primary tumor was removed at the time of OLT in 11 cases, by upper abdominal exenteration in 7 cases and the Whipple resection in 3. Actuarial survival rate after OLT was 59% at 1 year, 47% at 3 years, and 36% at 5 years. Survival rates were significantly higher for metastatic carcinoid tumors (69% at 5 years) than for noncarcinoid apudomas (8% at 4 years), because of higher tumor- and non-tumor-related mortality rates for the latter. CONCLUSION: OLT can achieve control of hormonal symptoms and prolong survival in selected patients with liver metastasis of carcinoid tumors. It does not seem indicated for other NET.

Adult↗
PubMed1996

The fetal specific gene CYP3A7 is inducible by rifampicin in adult human hepatocytes in primary culture.

We investigated by RNase protection the differential expression of CYP3A4 and CYP3A7 mRNAs in fetal and adult human livers and in adult hepatocytes in primary culture. Our results show that CYP3A7 is expressed in the liver of 8 of 9 adult Caucasians examined, at an average level of 1.7% of the level found in a fetal liver. CYP3A4 mRNA appeared to be expressed in this fetal liver. In adult hepatocytes in primary culture, the constitutive level of CYP3A4 mRNA was low but detectable after 96 hours in untreated cells, but CYP3A7 mRNA remained undetectable. However, when the cells were treated for 48 hours with 25 microM rifampicin, both CYP3A4 and CYP3A7 mRNAs were strongly induced in the 3 and in 2 of 3 cultures examined, respectively. Using an isoelectric focusing immunoblotting the two proteins were resolved. Protein CYP3A4 was detectable in induced cells but CYP3A7 was not. These results show for the first time that CYP3A7 and CYP3A4 mRNAs, but not the proteins, are co-inducible by rifampicin in adult human hepatocytes in culture.

Adult↗
PubMed1996

Metabolism of the new immunosuppressor cyclosporin G by human liver cytochromes P450.

Cyclosporin G is a new immunosuppressor structurally similar to cyclosporin A. Although this drug is pharmacologically as active as cyclosporin A, it is less toxic, in particular at the kidney level. The aim of this work was to identify the enzyme system(s) involved in the oxidative metabolism of cyclosporin G in man: (1) in a bank of human liver microsomes (n = 22), cyclosporin G oxidase activity correlated significantly with cyclosporin A oxidase activity (P < 0.0001) and with the level of CYP3A4 (P < 0.002), determined by immunoblot; (2) specific inhibitors of CYP3A4, troleandomycin, and ketoconazole, inhibited cyclosporin G oxidase activity by more than 80%; (3) antiCYP3A4 antibodies specifically inhibited this activity by nearly 90%; (4) cyclosporin A was a competitive inhibitor of cyclosporin G oxidase and vice versa; (5). Among a battery of cDNA-expressed CYPs, only CYP3A4 was able to generate detectable amounts of metabolites of cyclosporin G and cyclosporin A with a turnover number close to that calculated from experiments with liver microsomes; (6) in human hepatocytes in culture, pretreatment of cells with rifampicin and phenobarbital, 2 inducers of CYP3A4, produced a great increase in cyclosporin G oxidase activity, while beta-naphthoflavone, an inducer of CYP1As, did not. We conclude that CYP3A4 is the major enzyme involved in the oxidative metabolism of cyclosporin G in human liver.

Adult↗
PubMed1996

Auxiliary liver transplantation: regeneration of the native liver and outcome in 30 patients with fulminant hepatic failure--a multicenter European study.

Auxiliary liver transplantation (LT) is a special procedure of LT which could be proposed to patients with fulminant hepatic failure (FHF) and has for aim that complete regeneration of the native liver (NL) left in place will allow the graft recipient to resume normal liver function after allograft withdrawal. We report 30 cases of auxiliary LT performed for FHF in 12 European centers. Twenty-five of 30 patients were younger than 50 years. The cause of FHF was hepatitis A virus (HAV) in 4 patients, hepatitis B virus (HBV) in 7, paracetamol overdose in 5, ecstasy in 2, hepatotoxic drugs in 4, autoimmune hepatitis in 2, liver lesions of preeclampsia in 1 and unknown in 5. A postoperative, both clinical and histological follow-up of more than 3 weeks was obtained in 22 patients, enabling us to look for indicators predictive of NL regeneration and outcome. Histological changes observed in the NL included complete regeneration in 68%, incomplete regeneration with obvious fibrous sequelae in 14% and severe liver fibrosis or cirrhosis in 18%, of the 22 patients studied. The percentage and distribution of necrosis observed in tissue samples of the NL at the time of transplantation was not related to the final outcome. Complete NL regeneration was observed in 15 patients, out of whom 14 were younger than 40 years. Patients with complete regeneration were mainly affected by FHF due to HAV, HBV, or paracetamol overdose. After a follow-up of 18/11 (mean/median) months (range, 3 to 67 months), 19 of the 30 patients (63%) survived and 13 of them (68%), i.e., 43% of the 30 patients, had resumed normal NL function, with interrupted immunosuppression, the ultimate goal of emergency auxiliary LT. We conclude that, in patients with FHF, auxiliary LT is a procedure feasible in a number of centers and is associated with a complete regeneration capability of the NL in a majority of survivors, especially in those younger than 40 years. Confirmation of these encouraging preliminary results by large-scale prospective studies is required.

Adolescent↗
PubMed1996

Angiographic embolization of intrahepatic arterioportal fistula.

We report the case of a posttraumatic arteriovenous fistula between the right hepatic artery and the right portal vein remarkable in that clinical manifestations, including portal hypertension and mesenteric insufficiency findings, appeared latently and progressively worsened. This hepatoportal fistula was diagnosed by Doppler sonography and successfully treated by transcatheter embolization of feeding hepatic artery branch with steel coils. We emphasize the interest of pulsed Doppler in the diagnosis of hepatoportal fistula, in assessment of hemodynamic changes related to the fistula and in follow-up after treatment.

Angiography↗
PubMed1996

A phase II study of 5-fluorouracil, leucovorin and cisplatin (FLP) for metastatic gastric cancer.

The modulation of 5-fluorouracil (5-FU) with folinic acid (leucovorin, LV) is more efficacious than 5-FU alone in the treatment of metastatic colorectal cancer, and the combination of 5-FU with cisplatin is currently one of the most active regimens in advanced gastric cancer. A phase II study was therefore conducted to test the efficacy and toxicity of the combination of 5-FU, LV and cisplatin (FLP) in metastatic gastric cancer. 28 patients entered the study. Metastatic sites were observed in the liver (in 21 patients), the peritoneum (in 8), the lymph nodes (in 7) or the bones (in 1) and a local recurrence was noted in 4 cases. The performance status (using World Health Organisation criteria) was 0 for 13 patients and 1 or 2 for the others. Cycles of treatment were administered every 28 days and consisted of LV 200 mg/m2/day for 5 days followed by 5-FU 400 mg/m2/day for 5 days with cisplatin 100 mg/m2 on day 2. The response rate for the 27 evaluable patients was 51.8% (95% confidence interval (CI), 33-70.6%). There were four complete responses (14.8%) and 10 partial responses (37%). Median survival was 11 months and 4 patients were alive at 2 years. Both response rate and survival were better for patients with a good performance status. The overall toxicity was very low, except for 1 patient who died of dehydration and cardiac failure. In conclusion, the FLP protocol was effective and well tolerated in patients with metastatic gastric cancer.

Adult↗
PubMed1996

[Value of percutaneous hepatic biopsy in the diagnosis of presumed benign tumors of the liver].

The purpose of this study was to assess the accuracy of guided hepatic biopsy (GHB) and imaging techniques for presumed benign liver tumours and to determine their impact on surgical treatment. The study was carried out retrospectively in a surgical series of 15 consecutive patients with presumed benign liver tumours. The final diagnosis was 8 cases of focal nodular hyperplasia (FNH), 6 hepatic adenomas (HA) and one association FNH-HA. No morbidity was related to guided hepatic biopsy. All FNH detected on radiologic imaging or pathological examination of the biopsy specimen were true positive diagnoses. This study demonstrates that combined results of imaging techniques and percutaneous GHB could correctly diagnose three quarters of FNH before surgery. GHB is also useful when MRI imaging is indeterminate allowing a conservative approach for undiagnosed FNH.

Adenoma, Liver Cell↗
PubMed1995

[Gastroesophageal reflux. 2nd indication for digestive celioscopic surgery?].

Widespread us of laparoscopic surgery adds a new element to the debate over the choice between medical or surgical treatment for gastro-oesophageal reflux. Patients and gastroenterologists often favour medical management to avoid post-operative pain, a long recovery period, an abdominal scar or the risk of eventration but at the cost of long-term drug therapy and the need for repeated check-ups and endoscopy examinations. Yet surgery was found to give better long-term results in the only study comparing surgery and medical treatment. The question of cost and insurance coverage must also be considered. Although the indications for laparoscopic surgery would be identical to those for laparotomy it is probably possible that a wider population could benefit from this technique as laparoscopic cure can be indicated as an alternative in patients highly dependent on drug therapy. Relapse in patients with a long-term medical regimen is another recent indication. We should however always keep in mind that good outcome after laparoscopic surgery depends not only on a correct indication and evaluation of the oesophageal motricity but also on the skill and experience of the surgical team. Laparoscopic cure should certainly play a major role in the treatment of gastro-oesophageal reflux and will undoubtedly soon be the second most frequent laparoscopic technique performed after laparoscopic cholecystectomy.

Digestive System Surgical Procedures↗
PubMed1995

Tracheoesophagogastric anastomosis fistula: a rare complication of esophagectomy.

Fistulas between trachea and esophagogastric anastomosis after esophagectomy are uncommon. We describe 2 patients with such a lesion successfully managed with single-stage repair. The fistula was divided, the tracheal defect was closed directly or with a free pericardial graft, and the esophagogastric anastomosis was redone. A muscle or pleural flap was used to separate the tracheal and digestive suture lines. This treatment of a potentially life-threatening condition yielded excellent results without postoperative complications.

Adult↗
PubMed1995

Failure of ursodeoxycholic acid to prevent acute cellular rejection after liver transplantation.

BACKGROUND/AIMS: Acute rejection is still a major problem after liver transplantation. Ursodeoxycholic acid has beneficial effects in cholestasis by reducing the expression of major histocompatibility complex antigens. METHODS: We have performed a double-blind randomised study comparing ursodeoxycholic acid with placebo for the prevention of acute cellular rejection after liver transplantation. Twenty-six patients received ursodeoxycholic acid 600 mg per day and 24 patients received placebo for 2 months. RESULTS: Neither rejection incidence nor rejection severity was significantly different in the two groups (p > 0.90). CONCLUSIONS: We conclude that adjuvant ursodeoxycholic acid administration does not prevent rejection after liver transplantation.

Acute Disease↗