AIM To determine the influence of the construction design over the biological component's performance in an experimental bio-artificial liver(BAL) device.METHODS Two BAL models for liver microorgans(LMOs) were con...AIM To determine the influence of the construction design over the biological component's performance in an experimental bio-artificial liver(BAL) device.METHODS Two BAL models for liver microorgans(LMOs) were constructed. First, we constructed a cylindrical BAL and tested it without the biological component to establish its correct functioning. Samples of blood and biological compartment(BC) fluid were taken after 0, 60, and 120 min of perfusion. Osmolality, hematocrit, ammonia and glucose concentrations, lactate dehydrogenase(LDH) release(as a LMO viability parameter), and oxygen consumption and ammonia metabolizing capacity(as LMO functionality parameters) were determined. CPSI and OTC gene expression and function were measured. The second BAL, a "flat bottom" model, was constructed using a 25 cm2 culture flask while maintaining all other components between the models. The BC of both BALs had the same capacity(approximately 50 cm3) and both were manipulated with the same perfusion system. The performances of the two BALs were compared to show the influence of architecture.RESULTS The cylindrical BAL showed a good exchange of fluids and metabolites between blood and the BC, reflected by the matching of osmolalities, and glucose and ammonia concentration ratios after 120 min of perfusion. No hemoconcentration was detected, the hematocrit levels remained stable during the whole study, and the minimal percentage of hemolysis(0.65% ± 0.10%) observed was due to the action of the peristaltic pump. When LMOs were used as biological component of this BAL they showed similar values to the ones obtained in a Normothermic Reoxygenation System(NRS) for almost all the parameters assayed. After 120 min, the results obtained were: LDH release(%): 14.7 ± 3.1 in the BAL and 15.5 ± 3.2 in the NRS(n = 6); oxygen consumption(μmol/min?g wet tissue): 1.16 ± 0.21 in the BAL and 0.84 ± 0.15 in the NRS(n = 6); relative expression of Cps1 and Otc: 0.63 ± 0.12 and 0.67 ± 0.20, respectively, in the BAL, and 0.86 ± 0.10 and 0.82 �展开更多
Two specific carbamyl phosphate synthetase Ⅰ gene binding nuclear proteins (M. W. 109kD and 74 kD) have been determined in the rat liver by the protein blotting technique (South-western blot assay). The result shows ...Two specific carbamyl phosphate synthetase Ⅰ gene binding nuclear proteins (M. W. 109kD and 74 kD) have been determined in the rat liver by the protein blotting technique (South-western blot assay). The result shows that they are not present in the normal rat spleenand F-26 rat hepatoma cell. The Ba131 nuclease deletion in the CPSI gene 5’ upstream regionproves that the binding sites for 109 kD and 74 kD are respectively located in the regions of- 38 bp to - 4 bp and - 113 bp to -38 bp. The binding proteins may be the liver-specific onesof the CPSI gene, which are related to hepatocyte differentiation and hepatocarcinogenesis.展开更多
基金Supported by Universidad Nacional de Rosario(UNR),BIO 272,Resol.C.S.,No.677/2013Agencia Nacional de Promoción Científica y Tecnológica(ANPCyT),PICT-03-14492,BID 1728 OC/AR(Argentina)a grant from Regione Autonoma FriuliVenezia Giulia,Italy
文摘AIM To determine the influence of the construction design over the biological component's performance in an experimental bio-artificial liver(BAL) device.METHODS Two BAL models for liver microorgans(LMOs) were constructed. First, we constructed a cylindrical BAL and tested it without the biological component to establish its correct functioning. Samples of blood and biological compartment(BC) fluid were taken after 0, 60, and 120 min of perfusion. Osmolality, hematocrit, ammonia and glucose concentrations, lactate dehydrogenase(LDH) release(as a LMO viability parameter), and oxygen consumption and ammonia metabolizing capacity(as LMO functionality parameters) were determined. CPSI and OTC gene expression and function were measured. The second BAL, a "flat bottom" model, was constructed using a 25 cm2 culture flask while maintaining all other components between the models. The BC of both BALs had the same capacity(approximately 50 cm3) and both were manipulated with the same perfusion system. The performances of the two BALs were compared to show the influence of architecture.RESULTS The cylindrical BAL showed a good exchange of fluids and metabolites between blood and the BC, reflected by the matching of osmolalities, and glucose and ammonia concentration ratios after 120 min of perfusion. No hemoconcentration was detected, the hematocrit levels remained stable during the whole study, and the minimal percentage of hemolysis(0.65% ± 0.10%) observed was due to the action of the peristaltic pump. When LMOs were used as biological component of this BAL they showed similar values to the ones obtained in a Normothermic Reoxygenation System(NRS) for almost all the parameters assayed. After 120 min, the results obtained were: LDH release(%): 14.7 ± 3.1 in the BAL and 15.5 ± 3.2 in the NRS(n = 6); oxygen consumption(μmol/min?g wet tissue): 1.16 ± 0.21 in the BAL and 0.84 ± 0.15 in the NRS(n = 6); relative expression of Cps1 and Otc: 0.63 ± 0.12 and 0.67 ± 0.20, respectively, in the BAL, and 0.86 ± 0.10 and 0.82 �
基金Project supported by the National Natural Science Foundation of China.
文摘Two specific carbamyl phosphate synthetase Ⅰ gene binding nuclear proteins (M. W. 109kD and 74 kD) have been determined in the rat liver by the protein blotting technique (South-western blot assay). The result shows that they are not present in the normal rat spleenand F-26 rat hepatoma cell. The Ba131 nuclease deletion in the CPSI gene 5’ upstream regionproves that the binding sites for 109 kD and 74 kD are respectively located in the regions of- 38 bp to - 4 bp and - 113 bp to -38 bp. The binding proteins may be the liver-specific onesof the CPSI gene, which are related to hepatocyte differentiation and hepatocarcinogenesis.
