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Background/aims: The aim of this study was to assess the usefulness of the monoethylglycinexylidide (MEGX) test to monitoring the long-term function of liver allografts., Methods: MEGX production was measured prospectively in 60 consecutive liver transplant recipients undergoing their annual review., Results: Median MEGX values in liver recipients (54 ng/mL; range 10-146) were lower than those found in healthy controls (78 ng/mL; range 44-118). MEGX values correlated negatively with alanine aminotransferase (ALT) activity (p = 0.004) and with the overall histological score (p = 0.01), and positively with sulfobromophthalein (BSP) and indocyanine green (ICG) clearances (p = 0.0002 and p = 0.002, respectively). A stepwise decline was observed with worsening liver fibrosis, from 71 +/- 5 microg/L in patients with no fibrosis to 27 +/- 9 microg/L in patients with bridging fibrosis (p = 0.002). BSP and ICG clearances correlated more closely than the MEGX test with the overall histological score (p = 0.001 and p = 0.001, respectively) and portal fibrosis (p = 0.002 and p = 0.001)., Conclusions: The measurement of MEGX formation is a simple and non-invasive method to monitor liver graft function. It may constitute a valuable tool for assessing the degree of fibrosis.

BACKGROUND: After liver transplantation, the daily cyclosporine dose is adjusted to maintain the blood level in a chosen range. The aim of this study was to assess the influence of liver graft injury on cyclosporine dose. METHODS: The parameters of liver function were investigated in 145 patients. Ninety-two patients took part in a longitudinal study. RESULTS: The cyclosporine dose correlated with the MEGX test (r=0.38, P=0.01) and with the ICG (r=0.38, P=0.0001) and BSP (r=0.37, P=0.0002) clearances; it had an inverse correlation with transaminases (AST: r=0.38, P=0.0001) and histological lesions (r=-0.29, P=0.005). The cyclosporine dose was lower in patients with recurrent hepatitis C (179+/-9 mg/day) than in those without (241+/-10 mg/day), and was lower in patients with chronic hepatitis (154+/-9 mg/day) than in those without (207+/-16 mg/day). In the longitudinal study, the percent variation of AST correlated inversely with that of cyclosporine dose (r=-0.62, P=0.0002). CONCLUSION: Progressive graft injury leads to a reduction in the cyclosporine dose, particularly in patients with recurrent hepatitis C.

Objective: Systemic sclerosis (SSc) is characterized by fibrosis involving the skin and various internal organs. Type I collagen (Col I) is the most abundant extracellular matrix protein deposited in cutaneous involvement. We investigated Col I biochemical markers in patients with SSc., Methods: All consecutive patients admitted for SSc over a 9 month period and a healthy control group were investigated. Serum concentrations of the C-terminal telopeptide of Col I (s-CTX-I), C-terminal type I procollagen propeptide (PICP), osteocalcin, and bone alkaline phosphatases (ALP), and urinary concentrations of deoxypyridinoline (u-DPD) and u-CTX-I were determined by ELISA., Results: A total of 33 patients with SSc were included: mean age +/- SD was 54 +/- 12 yrs, with a mean disease duration of 5.4 +/- 3.9 years. Sixteen of 33 patients with SSc had s-CTX-I values exceeding the upper limit of normal values of the test and the mean +/- SEM was significantly higher (7388 +/- 1422 pmol/l) than in healthy controls (2800 +/- 1120 pmol/l; p < 0.001). s-CTX-I correlated with the Rodnan skin score (p = 0.003); it was higher in patients with diffuse disease (8459 +/- 3125; n = 14) than in patients with the limited form (6453 +/- 1235; n = 19; p < 0.02). This marker also correlated with acute phase reactants (C-reactive protein, p = 0.004; erythrocyte sedimentation rate, p = 0.02). s-CTX-I was high in patients with positive antitopoisomerase I autoantibodies (p = 0.04) and in patients with a decrease in forced vital capacity to less than 75% (p = 0.02). u-DPD concentration was high in patients with SSc (10.6 +/- 1.4 nmol/mmol creatinine vs 6.3 +/- 2.1 in controls; p < 0.01). No difference between patients and controls or correlations with the disease were found for PICP, u-CTX-I, osteocalcin, and bone ALP concentrations., Conclusion: s-CTX-I, a marker of Col I degradation, is correlated with cutaneous and pulmonary involvement and with acute phase reactants in patients with SSc. This marker is a good candidate for further evaluation for disease activity and treatment purposes.

The role of endothelin in the initial vasoconstrictor step of hyperacute xenogeneic rejection was investigated. Isolated rat livers were perfused in recirculation. Perfusion with human sera provided an ex vivo model of hyperacute rejection in a discordant combination. Perfusion of 10% xenogeneic serum induced a marked (70%) and sustained reduction of the liver flow and induced the release of endothelin into the perfusion medium. In contrast, perfusion of 10% allogeneic serum or of 10% decomplemented human serum induced a weak (25%) and transient reduction of the liver flow and induced the release of minimal amounts of endothelin. The simultaneous administration of BQ 123 and BQ 788, the respective antagonists of ET(A) and ET(B) endothelin receptors, or that of bosentan, a mixed ET(A)/ET(B) antagonist, antagonized the vasoconstrictor effect of 10% xenogeneic human serum, as well as that of 10(-9) M endothelin-1. The vasoconstrictor effects of xenogeneic serum on liver circulation are, at least partly, mediated through the release of endothelin by the graft.

Objective: To study the prevalence of osteoporosis (OP) and osteopenia in ankylosing spondylitis (AS) and to investigate the relationship between symptomatic and structural severity, the indices of bone turnover, and body composition., Methods: Eighty patients with AS were enrolled prospectively: 52 men (65%) and 28 women, mean age 36.7 years +/- 11.5 (range 18-67); they were studied clinically, radiologically, and by dual energy x-ray absorptiometry. Sixty-three underwent biological assessment of bone turnover markers., Results: OP and osteopenia as defined by the World Health Organization (T score < -2.5 SD and between -1 and -2.5 SD, respectively) were observed in 15 (18.7%) and 25 patients (31.2%) at the lumbar spine and in 11 (13.7%) and 33 patients (41.2%) at the femoral neck, respectively. Patients with OP had a lower body mass index (BMI) and fat mass percentage. There was a trend to a lower disease duration in patients with OP at the spine than in healthy subjects. Bone resorption markers (urinary D-pyridinoline or C-telopeptide concentrations) were increased in 34 patients (53.9%). Bone turnover markers were positively correlated with C-reactive protein concentration and Larsen radiological hip score; they were negatively correlated with Schober index and fat mass percentage., Conclusion: (1) OP is frequent in AS and can be observed in early stages of the disease. (2) Patients with AS are more susceptible to develop OP when they have low BMI, low fat mass percentage, and active and severe disease. OP was observed in parallel with increased bone resorption.

Three autologous blood units were transfused during elective orthopaedic surgery in a patient with undiagnosed haemoglobin SC disease. The packed red blood cells had been stored at 4 degrees C on SAG-M under standard conditions for 10 to 31 days. There was no evidence of adverse clinical reactions during the perioperative period. Six months later, a blood unit was collected at the initial step of an exchange transfusion in the same patient. Haemolysis was moderate after a 12-day-storage period and more significant after 32 days. This observation, as some other case reports, suggest that autologous blood transfusion may be considered for haemorrhagic surgery in selected patients with sickle cell disease.

Controlling the S phase of the hepatocyte cell cycle would be of considerable help for stable retroviral foreign gene transfer. The aim of this article is to study hepatocyte regeneration during S phase in isolated, perfused rat liver followed by liver transplantation. Normal livers (G I: n = 7) were perfused with blood from normal rats for 6.1+/-0.3 hours. Regenerating livers (G II; n = 7) obtained 18 hours after partial hepatectomy were perfused for 6.0+/-0.3 hours with blood from rats partially hepatectomized 18 hours before. Regenerating livers (G III; n = 7) obtained 22 hours after partial hepatectomy were perfused for 2.4+/-0.1 hours with blood from normal rats. In the normothermal perfusion system, a bolus of 25 mg of 5-bromo-2'-deoxyuridine (BrdU) was added to the perfusate. Liver biopsies were taken at the end of each experiment. In group II, a biopsy was also taken 1 hour after BrdU introduction. At the end of each experiment, livers were orthotopically transplanted. The percentage of BrdU positive hepatocyte nuclei was 0.2% in G I; 14.8% and 38.4% after 1 hour and 6.1 hours, respectively, in G II; and 46.5% after 2.4 hours in G III. In G I, five rats died at day 1, 5, 6, 7, and 48 and two rats were still alive after 17 months. In G II, all the rats died before day five. In G III, two rats died at day one, one at day six, and four were still alive after 12 months. This study shows that, after 6 hours of normothermal perfusion, organ viability allows successful liver transplantation and that rat hepatocyte regeneration during cell cycle S phase in isolated normothermal conditions progresses in a similar way-quantity and timing-to liver regeneration found in vivo after partial hepatectomy.

The method selected by the SFBC (Société française de biologie clinique) is derived from the colorimetric reaction of creatinine with alkaline picrate, measured kinetically, without any pretreatment step. The key parameters of the reaction determining the quality of the results are studied, with special regard to samples including known interferents. The aims of the study were to gain an optimal analytical sensitivity and to reduce main interferences (acetoacetate, bilirubine, glucose, protein) which plague the Jaffé reaction, through a comprehensive study of the reagents, of their concentrations and of the analytical procedures. The selected concentrations (in the test) are: 150 mmol/L sodium hydroxide, 10 mmol/L picric acid and 2 g/L sodium dodecyl sulfate. Ten millilitres of a BRIJ solution (30% volvol) are added to the reagent. The operating procedures are as follow: sample ratio 0.07 to 0.08; wavelength 505 to 510 nm; temperature 37 degrees C; incubation of the specimen with the alkaline reagent 5 mn (at least), before starting the reaction with picric acid. A seric calibrator is recommended. The first measurement is taken 20 to 40 s after starting the reaction. Total measurement time is 120 to 150 seconds.

A selected method for the determination of creatinine in plasma, using the reaction with alkaline picrate without prior pretreatment has been proposed by the Commission 'Validation de techniques' in the SFBC (Société Française de biologie clinique). The transferability step was conducted in seven laboratories, equipped with different automatic analyzers, using analytical procedures derived from the recommended method. Its goal was to test whether the original analytical performances could be maintained and consistent results obtained. The validation step was designed to evaluate the linearity limits of the analytical range, the detection limit, to assess accuracy as compared to a high performance liquid chromatography and to investigate the effect of the main interferents. Linearity limits are 15 and 2000 mumol/L. The detection limit is 3 to 8 mumol/L according to the analytical systems. The selected method can fulfil the set imprecision goals: intralaboratory CV minus than 2% (within-run), minus than 4% (run-to-run), interlaboratory CV minus than 5% (for 100 mumol/L creatinine). Inaccuracy evaluated for the chosen control sera is 1 to 15% as compared to the chromatographic method, according to the sera and to the analytical systems. The results obtained with the selected method are more consistent with the HPLC than are those obtained with an alkaline picrate method without SDS or with an enzymatic method. No interference could be demonstrated for acetoacetate (up to 8 mmol/L), hemoglobin (up to 210 mumol/L), unconjugated bilirubin (up to 250 mumol/L), glucose (up to 30 mmol/L), IgG (up to 45 g/L), albumin (up to 60 g/L). The effect of cephalosporins depends on the molecule. The reagents are stable for at least 6 months when stored in closed vials at +20 degrees C. The alkaline reagent is stable 30 days at +4 degrees C. Reference limits (0.025 and 0.975 fractiles) have been established for healthy adults. They are respectively 73 to 126 mumol/L for men and 59 to 100 mumol/L for females.

In cirrhotic livers, the intrahepatic resistance is increased and drug elimination and portal transhepatic flow are decreased. The aim of our work was to study the effect of a twofold increase in portal blood flow during 2 hr on the hemodynamic parameters, drug elimination and hepatic viability in eight isolated perfused human cirrhotic livers. Using an oxygenated recirculating system with independent arterial and portal flows, we perfused livers with Kreb's buffer bicarbonate solution, bovine serum albumin (20 gm.L-1) and human red blood cells (hematocrit 20%). The flow was maintained at a basal level of 0.713 +/- 0.19 L/min for 1 hr and then increased and maintained for 2 hr at twice the basal flow. Portal pressure-portal flow curve slopes were linear (27.04 +/- 21.06 mm Hg.L-1 x min; range = 6.43 to 60.8) and correlated with intrahepatic resistance during the basal-flow period (r = 0.87, p < 0.01). Parameters registered during the basal- and high-flow periods were compared by use of Student's t test: portal pressure increased from 23.5 +/- 7 to 37.3 +/- 16.7 mm Hg (p < 0.05); arterial pressure increased from 80.3 +/- 19 to 103.5 +/- 26 mm Hg (p < 0.005); hepatic artery flow resistance increased 31.9% (from 690.1 +/- 218 to 899.4 +/- 269 mm Hg.L-1 x min; p < 0.005); indocyanine green clearance increased by 28.2% (from 86.0 +/- 58.3 to 109.2 +/- 74.8 ml.min-1 x kg liver-1; p < 0.04). No significant differences were observed in enzyme release, biliary flow (n = 5) and oxygen consumption. Histological examinations demonstrated sinusoidal dilatations in six of eight cases.(ABSTRACT TRUNCATED AT 250 WORDS)

1. Isolated perfused cirrhotic rat livers were used to study the effects of an increase in portal perfusion pressure and portal flow on the microcirculation and viability of the hepatocytes. Cirrhosis was induced by CCl4, and Krebs-Ringer bicarbonate buffer solution was used as the perfusate. Portal perfusion pressures were increased incrementally between 25 and 45 cm H2O. The viability of the livers was assessed and histological studies were performed under light and electron microscopy. 2. An increase in portal perfusion pressure induced an increase in hepatic flow in all the experiments (P < 0.05). Hepatic flow was 2.52 ml min-1g-1 of liver (SD 0.67; n = 5) at basal pressure compared with 4.19 ml min-1g-1 of liver (SD 0.93; n = 5) and 5.91 ml min-1g-1 of liver (SD 0.63; n = 5) when pressures were raised to 25 and 45 cmH2O, respectively. Portal perfusion pressure and hepatic flow were correlated (r = 0.908; P < 0.001; n = 30). 3. Production of the enzyme alanine aminotransferase (EC 2.6.1.2) increased significantly from 5.69i.u. ml-1min g-1 of liver (SD 3.62; n = 5) to 23.53i.u. ml-1min g-1 of liver (SD 16.7; n = 5) when the perfusion pressure was raised from baseline to 30 cmH2O. In all the cases the porto-caval gradient of enzyme production was within the normal range. No correlation existed between the release of enzyme and portal perfusion pressures.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum delta-bilirubin measured in relationship to total bilirubin could be a liver function test in hepatobiliary diseases and even a marker of early rejection after liver transplantation. Delta-bilirubin bound to albumin constitutes with mono and diglucuroconjugates, conjugated bilirubins. We determined the delta-bilirubin/conjugated bilirubins ratio and studied its value in various complications in 16 liver recipients. The results were compared with alanine aminotransferase activity and prothrombin index. Hyperbilirubinemia and a delta-bilirubin/conjugated bilirubins ratio lower than 40% were observed initially. After 15 days, the delta-bilirubin/conjugated bilirubins ratio evolved differently according to the functional state of the graft, which allowed to distinguish three patient groups: in patients with good graft function, the ratio increased regularly to reach 65% or more on the 30th day; in patients with dysfunctional episodes, the ratio was always less than or equal to 40% and did not increase after the 15th day; in other patients, the ratio was lower than 40% and dropped suddenly before death. After liver transplantation, the delta-bilirubin/conjugated bilirubins ratio may be a simple liver function marker. Alone, however, it cannot be used to assess acute rejection.

During a multicenter evaluation, 16 methods for creatinine measurement have been tested according to the guidelines of the Société française de biologie clinique (SFBC) protocol. Kinetic Jaffé methods, widely used in France, performed on different analytical systems (Astra Beckman, IL 508, RA 1000 Technicon, Hitachi 704, 705, 717 Boehringer, Fara Roche, Progress Kone, Kem-O-Mat Coulter, Perspective France Monitor) have been compared to a continuous flow method with aqueous standards, to enzymatic methods using creatinine amidohydrolase with a colorimetric measurement (Boehringer and Ektachem Kodak) and to an HPLC method. Reproducibility, estimated with four different control sera, proved to be unsatisfactory in some cases as compared to current criteria for imprecision (less than +/- 10 mumol/l for intralaboratory and less than +/- 20 mumol/l for interlaboratory imprecision). The same selected patients sera covering the whole range of physiopathological concentrations have been analyzed with each method, and compared with the continuous flow results. Differences are more dependent on the sample than on the calibrators. The influences of haemolysis, bilirubin, acetoacetate, albumin, lipids, glucose, and some cephalosporins have been evaluated with spiked human sera. Haemolysed, turbid and jaundiced patient samples have been analyzed as well. The results vary according to the analytical procedure. This study took place in the implementation of a selected method for routine purpose with special regards to interferences and an acceptable imprecision. The method must satisfy the physicians' demands in the renal function exploration, especially in kidney-transplant patients.

Biotin deficiency was induced in germ-free rats using three experimental protocols. The results showed the important role of biotin during gestation and suckling. The earlier the deprivation, the earlier the deficiency and the severer the symptoms. In this vitamin deficiency, symptoms occurred that were not observed in the control rats, such as the formation of an intestinal volvulus in rats ingesting an L-isoleucine-supplemented diet. The main biochemical anomalies characteristic of propionic acidemia (ketoacidosis and increased urinary elimination of propionic acid) due to propionyl CoA carboxylase deficiency in man were not observed in the rats that were deprived of biotin for 200 days even after a dietary load of L-isoleucine. Only a small urinary excretion of propionyl-glycine and tiglylglycine was observed. We observed a drop in enzymatic propionyl CoA carboxylase activity in the liver that was proportional to the severity of the vitamin deficiency.

In this study we were able to evaluate the total lactate dehydrogenase activity of serum of children year by year. We also defined by electrophoresis the normal isozyme profile for each age group. We noted that the total activity was higher during the first years of life and that during this period, the isoenzymic profile was different from that of adults: higher percentage of LDH 5 (13% as against 8% in adults) and a lower percentage of LDH 1 (19% as against 24% in adults). Furthermore, it seemed to us preferable to determine the total LDH activity on fresh serum for a fall in activity is noted both after preservation at + 4 degrees C and -20 degrees C. Similarly, it is preferable to trace the isoenzyme profile on the same day rather than after preservation of the samples. The average differences show clearly the disappearance of certain fractions (isoenzymes 4 and 2 after six days preservation at + 4 degrees C). However, each serum seems to behave in a somewhat unforseeable manner and the dispersion of individual variations makes any statistical interpretation impossible.

Biotin deficiency induced in the rat, with a biotin-deficient diet but with added avidin involves clinical symptoms of deficiency as well as an important drop (80%) in propionyl-CoA carboxylase activity in the liver, heart and kidneys. But major biochemical anomalies (ketoacidosis and increased urinary elimination of propionic acid), characteristic of propionic acidaemia due to propionyl-CoA carboxylase deficiency in man, are not observed in the rat. Nevertheless, abnormal urinary elimination of methylcitrate and tiglylglycine reflects an appreciable decrease in the metabolism of propionyl-CoA in these animals. The propionyl-CoA load caused by the administration of metabolic precursors of this substance, mainly L-isoleucine, does not induce important biochemical variations except for excretion of propionylglycine.

An immuno-enzymatic assay of serum ferritin was performed in subjects over 70 years of age. Wide variations in serum ferritin levels were demonstrated in the non-anemic group (17-510 ng/ml). This makes interpretation of results very difficult. In anemic patients a negative correlation was found between serum ferritin levels and TIBC (Total Iron Binding Capacity). When TIBC is under 42 mumol/l or above 63 mumol/l the two values are so closely correlated that serum ferritin assay does not provide any additional information for the diagnosis of anemia. On the contrary when TIBC is between 42 and 63 mumol/l serum ferritin can allow an estimation of tissue iron storage particularly in absolute iron deficiencies.

Sodium dipropylacetate (DPA) was administered to 10 normally fed rats and 5 biotin-deficient rats. The effects of DPA were measured on propionic acid and glycine levels in urine and blood; and on hepatic propionyl-CoA carboxylase (PCC) activity. No abnormal amounts of propionic acid were found in the blood and urine of the treated rats. Hyperglycinemia and significant constant hyperglycinuria were observed in all animals given DPA, but without significant differences between normal and biotin-deprived rats. DPA treatment did not affect PCC activity in vivo or vitro. These results indicate that the abnormalities of the glycine metabolism are not linked to an accumulation of propionic acid due to PCC inhibition by DPA.

After administering sodium dipropylacetate to normal rats and biotin-deficient rats, we observed a slight hyperglycinemia, significant constant hyperglycinuria, but no accumulation of propionic acid in the blood or significant urinary elimination. Hepatic propionyl-CoA carboxylase activity, in vivo and in vitro, did not change, neither did sodium dipropylacetate inhibit propionyl-CoA carboxylase nor induce propionic acidemia resulting from a deficiency in this enzyme.