How nitrogen and phosphorus availability change water use efficiency in a Mediterranean savanna ecosystem

Nutrient availability, especially of nitrogen (N) and phosphorus (P), is of major importance for every organism and at a larger scale for ecosystem functioning and productivity. Changes in nutrient availability and potential stoichiometric imbalance due to anthropogenic nitrogen deposition might lead to nutrient deficiency or alter ecosystem functioning in various ways. In this study, we present 6 years (2014–2020) of flux-, plant-, and remote sensing data from a large-scale nutrient manipulation experiment conducted in a Mediterranean savanna-type ecosystem with an emphasis on the effects of N and P treatments on ecosystem-scale water-use efficiency (WUE) and related mechanisms. Two plots were fertilized with N (NT, 16.9 Ha) and N + P (NPT, 21.5 Ha), and a third unfertilized plot served as a control (CT). Fertilization had a strong impact on leaf nutrient stoichiometry only within the herbaceous layer with increased leaf N in both fertilized treatments and increased leaf P in NPT. Following fertilization, WUE in NT and NPT increased during the peak of growing season. While gross primary productivity similarly increased in NT and NPT, transpiration and surface conductance increased more in NT than in NPT. The results show that the NPT plot with higher nutrient availability, but more balanced N:P leaf stoichiometry had the highest WUE. On average, higher N availability resulted in a 40% increased leaf area index (LAI) in both fertilized treatments in the spring. Increased LAI reduced aerodynamic conductance and thus evaporation at both fertilized plots in the spring. Despite reduced evaporation, annual evapotranspiration increased by 10% (48.6 ± 28.3 kg H2O m−2), in the NT plot, while NPT remained similar to CT (−1%, −6.7 ± 12.2 kgH2O m−2). Potential causes for increased transpiration at NT could be increased root biomass and thus higher water uptake or rhizosphere priming to increase P-mobilization through microbes. The annual net ecosystem exchange shifted from a carbon source in CT (75.0 ± 20.6 gC m−2) to carbon-neutral in both fertilized treatments [−7.0 ± 18.5 gC m−2 (NT) 0.4 ± 22.6 gC m−2 (NPT)]. Our results show, that the N:P stoichiometric imbalance, resulting from N addition (without P), increases the WUE less than the addition of N + P, due to the strong increase in transpiration at NT, which indicates the importance of a balanced N and P content for WUE

Mutation analysis of SDHB and SDHC: novel germline mutations in sporadic head and neck paraganglioma and familial paraganglioma and/or pheochromocytoma

BACKGROUND: Germline mutations of the SDHD, SDHB and SDHC genes, encoding three of the four subunits of succinate dehydrogenase, are a major cause of hereditary paraganglioma and pheochromocytoma, and demonstrate that these genes are classic tumor suppressors. Succinate dehydrogenase is a heterotetrameric protein complex and a component of both the Krebs cycle and the mitochondrial respiratory chain (succinate:ubiquinone oxidoreductase or complex II). METHODS: Using conformation sensitive gel electrophoresis (CSGE) and direct DNA sequencing to analyse genomic DNA from peripheral blood lymphocytes, here we describe the mutation analysis of the SDHB and SDHC genes in 37 patients with sporadic (i.e. no known family history) head and neck paraganglioma and five pheochromocytoma and/or paraganglioma families. RESULTS: Two sporadic patients were found to have a SDHB splice site mutation in intron 4, c.423+1G>A, which produces a mis-spliced transcript with a 54 nucleotide deletion, resulting in an 18 amino acid in-frame deletion. A third patient was found to carry the c.214C>T (p.Arg72Cys) missense mutation in exon 4 of SDHC, which is situated in a highly conserved protein motif that constitutes the quinone-binding site of the succinate: ubiquinone oxidoreductase (SQR) complex in E. coli. Together with our previous results, we found 27 germline mutations of SDH genes in 95 cases (28%) of sporadic head and neck paraganglioma. In addition all index patients of five families showing hereditary pheochromocytoma-paraganglioma were found to carry germline mutations of SDHB: four of which were novel, c.343C>T (p.Arg115X), c.141G>A (p.Trp47X), c.281G>A (p.Arg94Lys), and c.653G>C (p.Trp218Ser), and one reported previously, c.136C>T, p.Arg46X. CONCLUSION: In conclusion, these data indicate that germline mutations of SDHB and SDHC play a minor role in sporadic head and neck paraganglioma and further underline the importance of germline SDHB mutations in cases of familial pheochromocytoma-paraganglioma

Piauí de plantas e gentes: Construção de coleções de referência de plantas úteis/econômicas como base para estudos arqueobotânicos.

Museum Studie

No evidence for involvement of SDHD in neuroblastoma pathogenesis

BACKGROUND: Deletions in the long arm of chromosome 11 are observed in a subgroup of advanced stage neuroblastomas with poor outcome. The deleted region harbours the tumour suppressor gene SDHD that is frequently mutated in paraganglioma and pheochromocytoma, which are, like neuroblastoma, tumours originating from the neural crest. In this study, we sought for evidence for involvement of SDHD in neuroblastoma. METHODS: SDHD was investigated on the genome, transcriptome and proteome level using mutation screening, methylation specific PCR, real-time quantitative PCR based homozygous deletion screening and mRNA expression profiling, immunoblotting, functional protein analysis and ultrastructural imaging of the mitochondria. RESULTS: Analysis at the genomic level of 67 tumour samples and 37 cell lines revealed at least 2 bona-fide mutations in cell lines without allelic loss at 11q23: a 4bp-deletion causing skip of exon 3 resulting in a premature stop codon in cell line N206, and a Y93C mutation in cell line NMB located in a region affected by germline SDHD mutations causing hereditary paraganglioma. No evidence for hypermethylation of the SDHD promotor region was observed, nor could we detect homozygous deletions. Interestingly, SDHD mRNA expression was significantly reduced in SDHD mutated cell lines and cell lines with 11q allelic loss as compared to both cell lines without 11q allelic loss and normal foetal neuroblast cells. However, protein analyses and assessment of mitochondrial morphology presently do not provide clues as to the possible effect of reduced SDHD expression on the neuroblastoma tumour phenotype. CONCLUSIONS: Our study provides no indications for 2-hit involvement of SDHD in the pathogenesis of neuroblastoma. Also, although a haplo-insufficient mechanism for SDHD involvement in advanced stage neuroblastoma could be considered, the present data do not provide consistent evidence for this hypothesis

Murine Models and Cell Lines for the Investigation of Pheochromocytoma: Applications for Future Therapies?

Pheochromocytomas (PCCs) are slow-growing neuroendocrine tumors arising from adrenal chromaffin cells. Tumors arising from extra-adrenal chromaffin cells are called paragangliomas. Metastases can occur up to approximately 60% or even more in specific subgroups of patients. There are still no well-established and clinically accepted “metastatic” markers available to determine whether a primary tumor is or will become malignant. Surgical resection is the most common treatment for non-metastatic PCCs, but no standard treatment/regimen is available for metastatic PCC. To investigate what kind of therapies are suitable for the treatment of metastatic PCC, animal models or cell lines are very useful. Over the last two decades, various mouse and rat models have been created presenting with PCC, which include models presenting tumors that are to a certain degree biochemically and/or molecularly similar to human PCC, and develop metastases. To be able to investigate which chemotherapeutic options could be useful for the treatment of metastatic PCC, cell lines such as mouse pheochromocytoma (MPC) and mouse tumor tissue (MTT) cells have been recently introduced and they both showed metastatic behavior. It appears these MPC and MTT cells are biochemically and molecularly similar to some human PCCs, are easily visualized by different imaging techniques, and respond to different therapies. These studies also indicate that some mouse models and both mouse PCC cell lines are suitable for testing new therapies for metastatic PCC