Regulation of endothelin-1 production in cultured rat mesangial cells

Regulation of endothelin-1 production in cultured rat mesangial cells. We investigated the regulatory mechanisms of endothelin (ET)-1 production in cultured rat mesangial cells (MC), with a special focus on the roles of protein kinase A (PKA)- and protein kinase C (PKC)-mediated signaling systems. Vasoactive agents and growth promoting factors, including platelet-derived growth factor, vasopressin and thrombin, which act through receptors coupled to the phospholipase C-mediated signaling system, as well as phorbol ester and fetal calf serum stimulated ET-1 production. This effect was attenuated in PKC-depleted or H-7 (a PKC inhibitor) treated MC. On the other hand, an increase in intracellular cyclic AMP by forskolin or β-adrenergic agonist, isoproterenol, which act as anti-mitogenic agents, inhibited serum-stimulated ET-1 production. In addition this effect was mimicked by the addition of 8-bromo-cyclic AMP to the medium. The effect of isoproterenol was abolished by propranolol. H-8, a PKA inhibitor, attenuated the inhibitory effect of forskolin. These findings suggest that ET-1 production in MC is regulated by interaction of both positive and negative signals mediated by PKC- and PKA-dependent mechanisms

Comparability of Weighed Dietary Records and a Self-Administered Diet History Questionnaire for Estimating Monetary Cost of Dietary Energy

An increasing number of studies have estimated monetary diet cost using various dietary assessment methods, based on databases on retail food prices, for investigating its association with dietary intake and health outcomes. However, information regarding the comparability of monetary diet cost across dietary assessment methods is absolutely lacking. This study compared monetary cost of dietary energy estimated from weighed dietary records (DRs) with that estimated from a self-administered diet history questionnaire (DHQ). The subjects were 92 Japanese women aged 31–69 years and 92 Japanese men aged 32–76 years. The DHQ (assessing diet during the preceding month) and 4-day DRs (one weekend day and three weekdays) were completed in each season over a 1-year period (DHQs1-4 and DRs1-4, respectively). An additional DHQ was completed at one year after completing DHQ1 (DHQ5). Monetary cost of dietary energy (Japanese yen/4184 kJ) was calculated using food intake information derived from each dietary assessment method, based on retail food prices. Pearson correlation between the mean of DRs1-4 and mean of DHQs1-4 was 0.64 for women and 0.69 for men. Pearson correlation between the mean of DRs1-4 and DHQ1 was 0.60 for women and 0.52 for men, while intraclass correlation between DHQ1 and DHQ5 was 0.64 for women and 0.51 for men. These data indicate reasonable comparability of monetary cost of dietary energy across DR and a DHQ as well as usefulness of a single administration of the DHQ for estimating monetary cost of dietary energy

Hemodynamic Analysis of a Microanastomosis Using Computational Fluid Dynamics

[Background] Technical issues in free flap transfer, such as the selection of recipient vessels and the positioning and method of anastomosis of the vascular pedicle, have been the subject of vigorous debate. Recent developments in computational fluid dynamics (CFD) have enabled the analysis of blood flow within microvessels. In this study, CFD was used to analyze hemodynamics in a microanastomosis. [Methods] In the fluid calculation process, the fluid domain modelizes microvessels with anastomosis. The inlet flow conditions were measured as venous waveform, and the fluid is simulated as blood. Streamlines (SL), wall shear stress (WSS), and oscillatory shear index (OSI) at the anastomosis were visualized and analyzed for observing effects from the flow field. [Results] Some flow disruption was evident as the SL passed over the sutures. The maximum recorded WSS was 13.37 Pa where the peak of a suture was exposed in the lumen. The local maximum value of the OSI was 0.182, recorded at the base of the anastomosis on the outflow side. [Conclusion] In the ideal anastomosis, the SL is disrupted as little as possible by the sutures. The WSS indicated that thrombus formation is unlikely to occur at suture peaks, but more likely to occur at the base of sutures, where the OSI is high. Tight suture knots are important in microanastomosis

CCN3 (NOV) Drives Degradative Changes in Aging Articular Cartilage

Aging is a major risk factor of osteoarthritis, which is characterized by the degeneration of articular cartilage. CCN3, a member of the CCN family, is expressed in cartilage and has various physiological functions during chondrocyte development, differentiation, and regeneration. Here, we examine the role of CCN3 in cartilage maintenance. During aging, the expression of Ccn3 mRNA in mouse primary chondrocytes from knee cartilage increased and showed a positive correlation with p21 and p53 mRNA. Increased accumulation of CCN3 protein was confirmed. To analyze the effects of CCN3 in vitro, either primary cultured human articular chondrocytes or rat chondrosarcoma cell line (RCS) were used. Artificial senescence induced by H2O2 caused a dose-dependent increase in Ccn3 gene and CCN3 protein expression, along with enhanced expression of p21 and p53 mRNA and proteins, as well as SA-beta gal activity. Overexpression of CCN3 also enhanced p21 promoter activity via p53. Accordingly, the addition of recombinant CCN3 protein to the culture increased the expression of p21 and p53 mRNAs. We have produced cartilage-specific CCN3-overexpressing transgenic mice, and found degradative changes in knee joints within two months. Inflammatory gene expression was found even in the rib chondrocytes of three-month-old transgenic mice. Similar results were observed in human knee articular chondrocytes from patients at both mRNA and protein levels. These results indicate that CCN3 is a new senescence marker of chondrocytes, and the overexpression of CCN3 in cartilage may in part promote chondrocyte senescence, leading to the degeneration of articular cartilage through the induction of p53 and p21