Evaluation of bioenergetic and mitochondrial function in liver transplantation

BACKGROUND/AIMS: We measured changes in mitochondrial function and bioenergetics that occur during ischemia/reperfusion in fresh liver samples of patients undergoing liver transplantation. These variations correlated with markers of liver function and clinical outcome. Ischemia/reperfusion injury related to liver transplantation affects mitochondrial function and bioenergetics. Experimental studies were conducted to identify the role of bioenergetics and mitochondrial dysfunction. To the best of our knowledge, no investigation of these two factors’ impacts on liver transplantation has been performed. METHODS: This was a prospective study of 28 patients who underwent liver transplantation. We measured parameters of mitochondrial function and bioenergetics in biopsies performed during the procedure. RESULTS: We observed a statistically significant reduction in mitochondrial membrane potential, an increase in lag phase, and decreases in mitochondrial respiration and adenosine triphosphate content (P<0.010). Higher postoperative aminotransferase peaks correlated with worse mitochondrial function; mitochondrial respiration correlated with arterial lactate (P<0.010). CONCLUSIONS: There is a relationship between mitochondrial function and ischemia/reperfusion injury. The future use of these clinical markers as prognostic factors may allow early identification of post-transplant liver failure and may indicate the need to perform a new transplant.

Construction of the dioxin bioassay method based on the clonal expressed aryl hydrocarbon receptor system / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the specific binding of the artificial clonal aryl hydrocarbon receptor translocator (ARNT) with the natural aryl hydrocarbon receptor (AhR) and the recolonization by polyclonal antibody. The dose-response relationship with tetrachlo-rodibenzo-dioxin (TCDD) was also studied to develop TCDD detection method and the binding degree related to dose response.</p><p><b>METHODS</b>(1) The target genes including AhR-PAS, AhR-C and ARNT-PAS were amplified by RT-PCR by using the total RNA purified from the liver cells of C57BL/6J mice as templates to construct pGEX-5X1 recombinants. The recombinant plasmids were expressed in E. coli. (2) The rabbits were immuned by the clonal fusion proteins: AhR-PAS, AhR-C to prepare the polyclonal antibody. (3) The natural AhR from the hepatic cytosol of C57BL/6J mice was extracted. The artificial cloning expressed fusion protein:GST-ARNT-PAS and the natural AhR were incubated in different dose of TCDD. The quantity of the heterodimer through affinity adsorption and Western blots were measured.</p><p><b>RESULTS</b>(1) The target proteins including AhR-PAS, AhR-C and ARNT-PAS were successfully cloned and expressed in E. coli. (2) The detection limit of polyclonal antibody AhR-PAS and AhR-C were 5 ng and 1 ng, respectively. (3) The total protein concentration prepared from the liver cells was 60.5 mg/ml. The artificial clonal protein ARNT-PAS could specifically bind to the natural AhR complex with the existence of TCDD. The detection limit of TCDD was 0.25 pmol which was 80 pg approximately.</p><p><b>CONCLUSION</b>A TCDD detection method based on the aryl hydrocarbon receptor system was established and the detection limit might reach pg grade.</p>

A multicenter clinical study of nafate for treatment of hepatic fibrosis in patients with chronic liver diseases / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the effect of nafate (compound fetal cow liver extract tablets) in hepatic fibrosis patients with chronic liver diseases.</p><p><b>METHODS</b>One hundred fifteen hepatic fibrosis patients with chronic liver diseases from 3 medical centers were included in this study. All patients were given nafate orally for twenty-four weeks. Before treatment and 12, 24 and 36 weeks after the treatment, serum levels of hyaluronic acid (HA), laminin (LN) and type IV collagen (IV-C) of the patients were measured by radioimmunoassay and their liver biopsy specimens were also assessed.</p><p><b>RESULTS</b>Before treatment and 24 and 36 weeks after the nafate treatment, serum levels of HA were (279.2+/-81.4) ng/ml, (136.8+/-56.7) ng/ml and (86.9+/-40.7) ng/ml respectively, serum levels of LN were (170.8+/-73.0) ng/ml, (112.5+/-39.5) ng/ml, and (60.8+/-31.8) ng/ml respectively, and IV-C levels were (153.7+/-60.1) ng/ml, (112.4+/-43.1) ng/ml, and (96.3+/-44.1) ng/ml respectively. There was a significant reduction in these serum values after the treatments. Histopathological examinations of the liver biopsies showed that the degree of hepatic fibrosis obviously declined by one or two degrees after the treatments.</p><p><b>CONCLUSIONS</b>nafate has positive effects in treating hepatic fibrosis.</p>

Peptone-liver [P-L] a new culture medium for the Egyptian leishmania major

Leishmania major promastigotes isolated from an Egyptian ZCL patient was inoculated 4x106/ml in a new culture medium, peptone-liver [P-L], with a total concentration of 10% of FCS [fetal calf serum] and incubated at 25 +/- 1C. Meanwhile, the same number of promastigotes was inoculated in Schneider's Drosophila medium as a control. On the 10th day, the number of L. major promastigotes reached 29.25x106/ml. The same number was obtained with Schneider's Drosophila medium within four days. The new culture medium or P-L medium is cheap, easily prepared and gave a very good number of promastigotes as in Schneider's Drosophila medium, but in a longer time

Effect of tissue and plasma factors on kidney proliferation

Liver extract, plasma from intact mice, ES2 tumour extract and plasma from tumour bearing mice has an inhibiting effect on the mitotic activity of hepatocytes and duodenal enterocytes. In the present experiments, the effect of these treatments on the mitotic activity of renal tubular cells was studied. C3HS 28 day-old male mice, standardized for periodicity analysis were used. The determination of normal mitotic circadian curve of the renocytes was done. A second batch of mice were injected with 0.01 ml/gr of either liver extract, plasma from intact mice, ES2 tumour extract or plasma from tumour bearing mice, at 16:00 hours and controlled at 08:00, 12:00 and 16:00 hs during 2 consecutive days post treatment. Colchicine (2 micrograms/gr) was injected 4 hours before killing. Kidneys were processed for histology and mitotic index determinations. Results were expressed as colchicine metaphases per 1000 nuclei, and showed that mitotic activity values of treated animals were significantly lower than those of controls. In conclusion, mitotic activity inhibition of renocytes may be due to some non specific plasmatic and/or tissue factors.

Accion del extracto de higado de ratones parcialmente hepatectomizados sobre la actividad mitotica de los enterocitos del raton joven / Action of liver extract from partially hepatectomized mice on the mitotic activity of young mouse enterocytes

En un trabajo anterior demonstramos que el plasma de retones adultos obtenido 36 horas post hepatectomía parcial, ejerce un efecto inhibitorio en la actividad mitótica de los enterocitos crípticos duodenales del ratón joven. En el presente trabajo se analiza la posibilidad de que dicho efecto se origine en algún factor del hígado regenerante. Para ello se estudia la acción del extracto de hígado de ratones adultos (90 días) obtenido 36 horas después de su hepatectomía parcial (70 por ciento), sobre la actividad mitótica de los enterocitos de ratones jóvenes, analizando 3 niveles celulares de las criptas duodenales. Se emplearon 36 ratones hembra de la cepa C3H/S de 27 días de edad la mitad de los cuales recibió, a las 16:00 horas, una inyección intraperitoneal de solución fisiológica, y restantes extracto hepático (0,01 ml/g). Lotes de 8 animales de cada grupo se sacrificaron a las 08:00/16, 12:00/20 y 16:00/24 (hora del día/horas post tratamiento) previa inyección de colchicina (2mug/g) 4 horas antes. Los resultados, expresados como metafases colchicínas por mil núcleos, demuenstran que la actividad mitótica, en los animales tratados con extracto, es significativamente menor con respecto a los testigos. El efecto inhibidor se manifesta en los niveles celulares de 1 a 4 y de 5 a 12 células de las criptas analizadas. En el nivel superior, de 13 a 20 células, no se aprecia ninguna modificación de la actividad proliferativa. Esta inhibición de la actividad mitótica de los enterocitos de las zonas basal y media de las criptas duodenales es probablemente debido a factores hepáticos difusibles.

Properties of channels from rat liver gap junction membrane fractions incorporated into planar lipid bilayers

Rat membrane fractions highly enriched for gap junctions can be incorporated into planar lipid bilayers exhibiting channel currents with both voltage-dependent and independent components. Voltage dependence, however, is only one of the characteristics of liver gap junction channels. Other features include poor ionic selectivity and sensitivity to calcium, pH, octanol and to some intracellularly applied antibodies. To further test the junctional nature of channels from membrane fractions highly enriched in gap junctions incorporated into lipid bilayers we studied the sensitivity of these channels to uncoupling agents and determined channel selectivity properties. We found the incorporated channels to be insensitive to calcium and octanol, and in most cases to pH in the range of 5-7, suggesting that either these agents do not interact directly with the junctional channels or that the corresponding gating regions are inactivated during the isolation and reconstitution procedures. Attempts to block channel activity using polyclonal and monoclonal connexin 32 antibodies were generally unsuccessful, although one antibody (a monoclonal directed against the carboxy terminus portion of connexin32) blocked channel activity. Selectivity measurements indicated that the incorporated channels were slightly cation selective (PNa=Pk > PCl) and were permeable to large ions. These results further support the idea that functional connexin32 gap junction channels are present in channel activity recorded from rat liver junctional membranes incorporated into planar bilayers

Modulation of lymphocyte proliferation by murine liver extract.

Murine liver extract (LEx) purified by ammonium sulfate (45-70% saturation) possesses a strong inhibitory effect on human lymphocyte proliferation. We have shown that the inhibitory effect of LEx is not via a cytotoxic effect and that it is proportional to the length of incubation with LEx. Mitogen-prestimulated lymphocytes are more resistant to LEx inhibition than cells not prestimulated. B cells stimulated by PWM are more susceptible to LEx-induced inhibition than PHA- or Con A-stimulated T cells. In Con A cultures, there may be a population of cells more resistant to LEx inhibition. This population is not yet identified. The degree of reversibility of LEx inhibition was different in cells prestimulated by different mitogens. The inhibitory activity of LEx decreased in the presence of an increasing number of cells in the culture.

Effects of steroid drugs on some chemical constituents of the liver of hamsters infected with schistosoma mansoni I Oral contraceptive

The present work deals with the effect of Nordette [oral contraceptive] on some chemical constituents of the liver of normal and Schistosoma mansoni infected hamsters. Infection was found to cause an increase in the weight and water content of the liver. On the other hand, a decrease in the glycogen content and succinate dehydrogenase activity was recorded. In case of Nordette treated hamsters, the lipids content was increased in the liver of normal hamsters, while succinate dehydrogenase activity was decreased in both normal and infected animals

Effects of steroid drugs on some chemical constituents of the liver of hamsters infected with schistosoma mansoni II Glucocorticoid

Betamethasone [9 alpha-fluoro-16 beta-methyl-prednisolone] was administered orally [20 ug/100 g/day for 7 days] to normal and Schistosoma mansoni infected hamsters. The drug increased the liver weight in normal hamsters, but no such effect was seen in infected animals. A decrease in the proteins content of liver was noted in both normal and infected hamsters after Betamethasone treatment. Also, a decrease in the hepatic lipids content was evident in infected hamsters but not in normals. In normal and infected hamsters treated with Betamethasone, there was no change in hepatic contents of water, glycogen and succinate dehydrogenase activity