Background/Aims: S-Adenosylhomocysteine hydrolase (AdoHcyase) catalyzes the reversible hydrolysis of S-adenosylhomocysteine (AdoHcy), which is a potent product inhibitor of S-adenosylmethionine (AdoMet)-dependent methyltransferases. While previous studies have shown that AdoHcyase inhibition or deficiency lead to a decreased AdoMet/AdoHcy ratio resulting in impaired transmethylation, the effect of enhanced AdoHcyase activity on AdoMet/AdoHcy metabolism and methylation reactions has not been studied in detail. Methods: To investigate the effect of enhanced AdoHcyase activity, we generated HEK-293 cell lines stably overexpressing AdoHcyase. Results: Initial studies revealed that 2-10-fold AdoHcyase overexpression resulted in decreased intracellular AdoHcy and elevated adenosine levels, whereas 16-fold AdoHcyase overexpression increased adenosine and AdoHcy levels, lowered energy charge, and altered cell morphology. Furthermore, we found a correlation between AdoHcyase activity and cell viability. Caspase-activity assays and DNA fragmentation analysis revealed that the cell death in AdoHcyase overexpressing cells was due to apoptosis. Global DNA methylation was not altered in the different AdoHcyase overexpressing cell lines. Conclusion: Taken together, these data show that 2-5-fold enhanced AdoHcyase activity is well tolerated by the cell, while greatly enhanced AdoHcyase activity results in adenosine-induced apoptosis. The fact that enhanced AdoHcyase activity does not increase transmethylation activity suggests that AdoHcyase activity under physiological conditions is not rate limiting for efficient transmethylation.

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Over the last 20 years, the species assemblage of the species-rich dry grassland communities of central Germany has changed due to the ongoing abandonment of traditional land-use practices. In our study we wanted to investigate the germination biology and the plant-plant interaction of the low-growing and declining forb Alyssum montanum and the increasingly dominant grass Festuca rupicola. To investigate the germination behaviour we simulated cold (8/4°C), warm (20/10°C) and hot (32/20°C) conditions under a light-dark regime (12/12 hours) as well as in constant darkness. Germination of both species was similar with almost all non-dormant seeds germinating under intermediate temperature conditions (20/10°C). Whereas F. rupicola (F) germinated equally well under light changing conditions and constant darkness, the germination of A. montanum (A) was clearly reduced in darkness, which implies a strong competitive advantage of F. rupicola in dense vegetation cover. However, A. montanum germinated early under all temperatures regimes and better under 20/10°C as well as with light changing conditions. The interactions between the two species were tested in a pod-experiment with a replacement design, within which the composition of individuals was changed in the following way: 4A/1F, 3A/2F, 2A/3F, 1A/4F (eight replications per each). To evaluate any influence of intraspecific competition we created monocultures in an additive design (1A, 2A, 3A, 4A, 5A, 1F, 5F; eight replications per each). Despite that A. montanum developed faster and flowered after only four months in the replacement design experiment, the herb responded negatively to the presence of F. rupicola in several parameters (number of flowers, number of fruits and biomass per individual). The calculation of the Relative Neighbour Effect (RNE) indicated facilitation for F. rupicola and competition on A. montanum, which increased with increasing number of individuals of the other species. Our data suggest that A. montanum seedling recruitment, growth and fitness are negatively affected by increasing vegetation density of the expanding grass species F. rupicola.

Glomerular filtration rate (GFR) is inversely and thus paradoxically related to dietary NaCl intake in rats and patients with early type 1 diabetes mellitus (DM). Enhanced sensitivity of proximal reabsorption to NaCl diet inducing secondary adaptations in GFR through actions of tubuloglomerular feedback causes this salt paradox. We studied the role of renal nerves for the salt paradox in rats with streptozotocin (STZ)-induced DM since a regulatory influence of renal nerves on proximal reabsorption is well established. The left kidney (LK) was denervated before induction of STZ-DM. Subsequently, the normal diet was continued or a low NaCl diet was initiated and 1 week later animals were prepared for clearance experiments under anesthesia including ureter catheterization to measure GFR for each kidney. In diabetic rats, the right innervated as well as the left denervated kidney showed higher values for GFR and kidney weight in animals on a low versus a normal NaCl diet indicating that the salt paradox occurs independent of renal innervation. In addition, evidence is provided that the renal nerves of non-diabetic rats do not contribute to renal Na⁺ retention during dietary NaCl restriction but modulate renal hemodynamics and kidney weight under these conditions.

The present study was designed to investigate, in human subjects, urinary dopamine excretion under different conditions of sodium and water homeostasis. In a cross-over trial, ten healthy volunteers were subjected to low-salt (LS; dietary salt restriction, sodium chloride (NaCl) intake <5 g per day), normal-salt (NS; normal food ad libitum), and high-salt (HS; normal food plus NaCl 100 mg/kg per day) regimens for 8 days in a randomized order. On day 7, urine was collected for 24 h. The variations in urinary sodium excretion reflected the dietary salt intake (LS: 16.3+/-4.7; NS: 144.1+/-18.2; HS: 221.9+/-12.9 mmol 24 h(-1) 1.73 m(-2)), but were not accompanied by significant changes in urinary dopamine excretion. On day 8, clearance studies showed that an acute oral water load of 1500 ml did not alter glomerular filtration rate or renal plasma flow but significantly increased urinary flow rate without affecting dopamine excretion. Assuming that excreted dopamine is not metabolized or reabsorbed during the tubular passage, both the unchanged urinary dopamine output in spite of 14-fold variations in sodium excretion and its independence of an acute water load argue against the hypothesis that dopamine in the tubular lumen acts as a natriuretic and/or diuretic factor in humans.