Enzyme kinetics inform about mechanistic changes in tea litter decomposition across gradients in land-use intensity in Central German grasslands.

Grassland ecosystems provide important ecosystem services such as nutrient cycling and primary production that are affected by land-use intensity. To assess the effects of land-use intensity, operational and sensitive ecological indicators that integrate effects of grassland management on ecosystem processes such as organic matter turnover are needed. Here, we investigated the suitability of measuring the mass loss of standardized tea litter together with extracellular enzyme kinetics as a proxy of litter decomposition in the topsoil of grasslands along a well-defined land-use intensity gradient (fertilization, mowing, grazing) in Central Germany. Tea bags containing either green tea (high-quality litter) or rooibos tea (low-quality litter) were buried in 5 cm soil depth. Litter mass loss was measured after three (early-stage decomposition) and 12 months (mid-stage decomposition). Based on the fluorescence measurement of the reaction product 4-methylumbelliferone, Michaelis-Menten enzyme kinetics (Vmax: potential maximum rate of activity; Km: substrate affinity) of five hydrolases involved in the carbon (C)-, nitrogen (N)- and phosphorus (P)-cycle (ß-glucosidase (BG), cellobiohydrolase (CBH), cellotriohydrolase (CTH), 1,4-ß-N-acetylglucosaminidase (NAG), and phosphatase (PH)) were determined in tea litter bags and in the surrounding soil. The land-use intensity index (LUI), summarizing fertilization, mowing, grazing, and in particular the frequency of mowing were identified as important drivers of early-stage tea litter decomposition. Mid-stage decomposition was influenced by grazing intensity. The higher the potential activity of all measured C-, N- and P-targeting enzymes, the higher was the decomposition of both tea litters in the early-phase. During mid-stage decomposition, individual enzyme parameters (Vmax of CTH and PH, Km of CBH) became more important. The tea bag method proved to be a suitable indicator which allows an easy and cost-effective assessment of land-use intensity effects on decay processes in manged grasslands. In combination with enzyme kinetics it is an appealing approach to identify mechanisms driving litter break down.

Ascorbic acid-induced TET activation mitigates adverse hydroxymethylcytosine loss in renal cell carcinoma.

Although clear cell renal cell carcinoma (ccRCC) has been shown to result in widespread aberrant cytosine methylation and loss of 5-hydroxymethylcytosine (5hmC), the prognostic impact and therapeutic targeting of this epigenetic aberrancy has not been fully explored. Analysis of 576 primary ccRCC samples demonstrated that loss of 5hmC was strongly associated with aggressive clinicopathologic features and was an independent adverse prognostic factor. Loss of 5hmC also predicted reduced progression-free survival after resection of nonmetastatic disease. The loss of 5hmC in ccRCC was not due to mutational or transcriptional inactivation of ten eleven translocation (TET) enzymes, but to their functional inactivation by l-2-hydroxyglutarate (L2HG), which was overexpressed due to the deletion and underexpression of L2HG dehydrogenase (L2HGDH). Ascorbic acid (AA) reduced methylation and restored genome-wide 5hmC levels via TET activation. Fluorescence quenching of the recombinant TET-2 protein was unaffected by L2HG in the presence of AA. Pharmacologic AA treatment led to reduced growth of ccRCC in vitro and reduced tumor growth in vivo, with increased intratumoral 5hmC. These data demonstrate that reduced 5hmC is associated with reduced survival in ccRCC and provide a preclinical rationale for exploring the therapeutic potential of high-dose AA in ccRCC.

Land-use and soil depth affect resource and microbial stoichiometry in a tropical mountain rainforest region of southern Ecuador.

Global change phenomena, such as forest disturbance and land-use change, significantly affect elemental balances as well as the structure and function of terrestrial ecosystems. However, the importance of shifts in soil nutrient stoichiometry for the regulation of belowground biota and soil food webs have not been intensively studied for tropical ecosystems. In the present account, we examine the effects of land-use change and soil depth on soil and microbial stoichiometry along a land-use sequence (natural forest, pastures of different ages, secondary succession) in the tropical mountain rainforest region of southern Ecuador. Furthermore, we analyzed (PLFA-method) whether shifts in the microbial community structure were related to alterations in soil and microbial stoichiometry. Soil and microbial stoichiometry were affected by both land-use change and soil depth. After forest disturbance, significant decreases of soil C:N:P ratios at the pastures were followed by increases during secondary succession. Microbial C:N ratios varied slightly in response to land-use change, whereas no fixed microbial C:P and N:P ratios were observed. Shifts in microbial community composition were associated with soil and microbial stoichiometry. Strong positive relationships between PLFA-markers 18:2n6,9c (saprotrophic fungi) and 20:4 (animals) and negative associations between 20:4 and microbial N:P point to land-use change affecting the structure of soil food webs. Significant deviations from global soil and microbial C:N:P ratios indicated a major force of land-use change to alter stoichiometric relationships and to structure biological systems. Our results support the idea that soil biotic communities are stoichiometrically flexible in order to adapt to alterations in resource stoichiometry.