Expression of carbamoylphosphate synthetase I and glutamine synthetase in hepatic organoids reconstructed by rat small hepatocytes and hepatic nonparenchymal cells.

The objective of this study was to investigate the expression of carbamoylphosphate synthetase I (CPS) and glutamine synthetase (GS) in small hepatocyte colonies and whether the heterogeneous expression of the enzymes could be induced during the maturation of small hepatocytes. Small hepatocytes isolated from an adult rat liver were cultured and proliferated to form colonies. The expression of CPS and GS was examined using immunocytochemistry and immunoblotting. In this culture more than 99% of morphologically hepatic cells were positive for CPS and all small hepatocytes were negative for GS at day 5. CPS-positive cells dramatically decreased with time in culture, whereas GS-positive ones appeared and their number increased in the colonies. Two to 3 weeks after plating, colonies with rising and piled-up cells appeared and the number of such colonies reached about 25% of all colonies at day 30. In most rising and piled-up cells in colonies both proteins were strongly expressed, whereas many small hepatocytes in monolayer colonies did not express either protein. When small hepatocytes in monolayer colonies were overlayed with Matrigel, the cells gradually piled up and both CPS and GS proteins were dramatically induced. The expression of CPS and GS in small hepatocytes may interact with the extracellular matrix because the rising and piled-up cells appear to be induced by the extracellular matrix produced by hepatic nonparenchymal cells.

Analysis of blood pressure responses during exercise by logistic function curve in hypertension: effects of age, gender and physical training.

During exercise blood pressure fluctuates from minute to minute and does not rise linearly with time. Blood pressure responses were evaluated during exercise by a logistic function curve. Thirty-nine hypertensive patients underwent exercise testing with an ergometer, employing a multistage method (25 watts increment, every 3 min). We plotted the exercise duration on the X-coordinate and systolic blood pressure on the Y-coordinate and blood pressure was assumed to form a logistic curve for exercise duration. The relationship of systolic blood pressure vs. exercise duration was better fitted into a logistic function curve than a linear regression model. The logistic curve was defined by lower plateau, upper plateau, SPX (the X-coordinate at the shift point) and df (SPX), the maximal slope at the shift point. The effects of aging, gender and physical training were then analyzed on the curve. Aging did not affect lower plateau, upper plateau and SPX but augmented df (SPX), indicating greater blood pressure responses in older subjects during exercise. In females the curve was shifted to the left compared to males (SPX: 4.9 vs. 8.3 min, P < 0.05) without changes in plateaux and df (SPX), indicating greater blood pressure responses than males. Physical training for 3 weeks decreased the lower plateau from 157 to 144 mmHg (P < 0.05) and shifted the curve to the right (SPX: 7.1 vs. 8.6 min, P < 0.05), indicating unchanged blood pressure responses after training because of the opposite effects by decreases in the lower plateau vs. the curve shift to the right. In conclusion, blood pressure during exercise is better delineated by a logistic function curve than a linear regression model. The biological or physiological significance of df (SPX) is not clear at present and needs further investigations.