Establishment of a mouse model with mutagenesis induced hyperaldosteronism

According to recent epidemiological studies, primary aldosteronism is considered to be responsible for almost 10-15% of all cases of arterial hypertension. The genetic background of this common disease, however, has been elucidated only for the rare familial types whereas in the large majority of sporadic cases genetic causes or modifiers still remain unclear. In an attempt to define novel genetic mechanisms of hyperaldosteronism we utilized a random mutagenesis screen after treatment with the alkylating agent N-ethylnitrosourea (ENU) and phenotypically characterized affected mice for their blood aldosterone levels. As the detection method we used a time resolved fluorescent immunoassay which allows the measurement of aldosterone in very small murine plasma volumes. Using this assay we determined the normal aldosterone values for C3HeB/FeJ wild type mice under baseline conditions and following specific stimulation and suppression tests. We could demonstrate the expected increase in aldosterone response upon ACTH stimulation, a high potassium diet and an angiotensin II injection, as well as the decrease after a dexamethasone suppression test, a normal saline load test and a fludrocortisone suppression test. On the molecular level, the expression of aldosterone synthase showed a similar pattern with a fast response to the investigated stimuli. These tests should later be applied to the mouse lines derived from the ENU screen, in order to investigate potential abnormal response to these stimuli in comparison to wild type animals. Furthermore, aldosterone measurement was carried out in more than 2000 F1 offspring (of both genders) of chemically mutated inbred C3HeB/FeJ mice. From these tested F1 offspring, aldosterone levels were consistently elevated (defined as levels above 3 SD over the mean of untreated animals)upon repeated measurement in 9 animals (8 females and 1 male). Further breeding of affected female animals gave rise to F2 pedigrees from which four established lines displayed high aldosterone values. These animals served for a detailed phenotypic characterization and showed an increased aldosterone to renin ratio, low potassium values and normal renal function in line with the presence of primary aldosteronism. In addition, the investigation of their cardiac phenotype showed increased collagen deposits and subsequently cardiac fibrosis, as also observed in patients suffering from primary aldosteronism. In the future, genetic SNP analysis can be performed to identify underlying genetic loci, responsible for this trait. Taken together, these data demonstrate the feasibility of a phenotype-driven mutagenesis screen to detect and establish mutant mouse lines with a high aldosterone phenotype

Transcriptomics, Epigenetics, and Metabolomics of Primary Aldosteronism

INTRODUCTION Primary aldosteronism (PA) is the most common cause of endocrine hypertension, mainly caused by aldosterone-producing adenomas or hyperplasia; understanding its pathophysiological background is important in order to provide ameliorative treatment strategies. Over the past several years, significant progress has been documented in this field, in particular in the clarification of the genetic and molecular mechanisms responsible for the pathogenesis of aldosterone-producing adenomas (APAs). METHODS Systematic searches of the PubMed and Cochrane databases were performed for all human studies applying transcriptomic, epigenetic or metabolomic analyses to PA subjects. Studies involving serial analysis of gene expression and microarray, epigenetic studies with methylome analyses and micro-RNA expression profiles, and metabolomic studies focused on improving understanding of the regulation of autonomous aldosterone production in PA were all included. RESULTS In this review we summarize the main findings in this area and analyze the interplay between primary aldosteronism and several signaling pathways with differential regulation of the RNA and protein expression of several factors involved in, among others, steroidogenesis, calcium signaling, and nuclear, membrane and G-coupled protein receptors. Distinct transcriptomic and metabolomic patterns are also presented herein, depending on the mutational status of APAs. In particular, two partially opposite transcriptional and steroidogenic profiles appear to distinguish APAs carrying a KCNJ5 mutation from all other APAs, which carry different mutations. CONCLUSIONS These findings can substantially contribute to the development of personalized treatment in patients with PA

A Bayesian Logistic Regression approach in Asthma Persistence Prediction

Background: A number of models based on clinical parameters have been used for the prediction of asthma persistence in children. The number and significance of factors that are used in a proposed model play a cardinal role in prediction accuracy. Different models may lead to different significant variables. In addition, the accuracy of a model in medicine is really important since an accurate prediction of illness persistence may improve prevention and treatment intervention for the children at risk. Methods: Data from 147 asthmatic children were analyzed by a new method for predicting asthma outcome using Principal Component Analysis (PCA) in combination with a Bayesian logistic regression approach implemented by the Markov Chain Monte Carlo (MCMC). The use of PCA is required due to multicollinearity among the explanatory variables. Results: This method using the most appropriate models seems to predict asthma with an accuracy of 84.076% and 86.3673%, a Sensitivity of 84.96% and 87.25% and a Specificity of 83.22% and 85.52%, respectively. Conclusion: Our approach predicts asthma with high accuracy, gives steadier results in terms of positive and negative patients and provides better information about the influence of each factor (demographic, symptoms etc.) in asthma prediction

Mouse models of primary aldosteronism: from physiology to pathophysiology

Primary aldosteronism (PA) is a common form of endocrine hypertension that is characterized by the excessive production of aldosterone relative to suppressed plasma renin levels. PA is usually caused by either a unilateral aldosterone-producing adenoma or bilateral adrenal hyperplasia. Somatic mutations have been identified in several genes that encode ion pumps and channels that may explain the aldosterone excess in over half of aldosterone-producing adenomas, whereas the pathophysiology of bilateral adrenal hyperplasia is largely unknown. A number of mouse models of hyperaldosteronism have been described that recreate some features of the human disorder although none replicate the genetic basis of human PA. Animal models that reproduce the genotype-phenotype associations of human PA are required to establish the functional mechanisms that underlie the endocrine autonomy and deregulated cell growth of the affected adrenal and for preclinical studies of novel therapeutics. Herein, we discuss the differences in adrenal physiology across species and describe the genetically-modified mouse models of PA that have been developed to date

Deoxyguanosine kinase mutation F180S is associated with a lean phenotype in mice

Background: Deoxyguanosine kinase (DGUOK) deficiency is one of the genetic causes of mitochondrial DNA depletion syndrome (MDDS) in humans, leading to the hepatocerebral or the isolated hepatic form of MDDS. Mouse models are helpful tools for the improvement of understanding of the pathophysiology of diseases and offer the opportunity to examine new therapeutic options. Methods: Herein, we describe the generation and metabolic characterization of a mouse line carrying a homozygous DguokF180S/F180S mutation derived from an N-ethyl-N-nitrosourea-mutagenesis screen. Energy expenditure (EE), oxygen consumption (VO2) and carbon dioxide production (VCO2) were assessed in metabolic cages. LC-MS/MS was used to quantify plasma adrenal steroids. Plasma insulin and leptin levels were quantified with commercially available assay kits. Results: Mutant animals displayed significantly lower body weights and reduced inguinal fat pad mass, in comparison to unaffected littermates. Biochemically, they were characterized by significantly lower blood glucose levels, accompanied by significantly lower insulin, total cholesterol, high density lipoprotein and triglyceride levels. They also displayed an almost 2-fold increase in transaminases. Moreover, absolute EE was comparable in mutant and control mice, but EE in mutants was uncoupled from their body weights. Histological examination of inguinal white adipose tissue (WAT) revealed adipocytes with multilocular fat droplets reminiscent of WAT browning. In addition, mRNA and protein expression of Ucp1 was increased. Mutant mice also presented differing mitochondrial DNA content in various tissues and altered metabolic activity in mitochondria, but no further phenotypical or behavioral abnormalities. Preliminary data imply normal survival of DguokF180S/F180S mutant animals. Conclusion: Taken together, DGUOK mutation F180S leads to a lean phenotype, with lower glucose, insulin, and lipid levels rendering this mouse model not only useful for the study of MDDS forms but also for deciphering mechanisms resulting in a lean phenotyp

Carbon sequestration and soil nitrogen enrichment in Robinia pseudoacacia L. post-mining restoration plantations

Robinia pseudoacacia L. (black locust) has been extensively used for restoring degraded lands, following anthropogenic interventions like coal mining. Here we have addressed the contribution of black locust restoration plantations, established on overburden post-mining material, to carbon storage and to soil nitrogen enrichment at the largest lignite center in Greece. Carbon stocks and fluxes in all pools of the ecosystem, as well as the foliar nitrogen resorption efficiency and soil N stocks were quantified and the effect of plantations’ age was tested. The young age of the plantations (4–24 years) resulted in a relatively low total ecosystem C stock (56.7 t ha−1), which was partitioned among the different pools in the following order: above-ground biomass (50%) > black locust-derived SOC (24%) > coarse roots (14%) > deadwood (6%) > forest floor (5%) > fine roots (less than 1%). Litterfall started early in the growing season and together with fine roots that had a turnover rate of 0.62 yr−1, fueled soil organic carbon. SOC accrual, referring to the accumulation of SOC derived by black locust, declined with age. However, further SOC accumulation is expected, based on the potential SOC storage capacity of soil at the area. C stocks in above- and below-ground biomass increased linearly with age. The same response was observed for soil N stock and NRE, indicating that despite the N2-fixing capacity of black locust, there was still a poor pedospheric N supply and a need for efficient N cycling. Overall, the studied restoration plantations have a considerable contribution to C and N accumulation at the degraded post-mining sites. These positive effects are expected to further increase at least until the plantations reach maturity

Statins as antifungal agents

This poster paper describes the objectives, approach and use casesof the EC FP7 Integrated Project PERICLES. The project beganon 1st February 2013 and runs for four years. The aim is toresearch and prototype solutions for digital preservation incontinually evolving environments including changes in context,semantics and practices. The project addresses use cases focusingon digital art, media and science.Proceedings source: http://purl.pt/24107/1/iPres2013_PDF/iPres2013-Proceedings.pd

Impact of 123 I-MIBG scintigraphy on clinical decision making in pheochromocytoma and paraganglioma

CONTEXT Cross sectional imaging with computed tomography (CT) or magnetic resonance imaging (MRI) is regarded as a first-choice modality for tumor localization in patients with pheochromocytoma and paraganglioma (PPGL). 123I-labeled metaiodobenzylguanidine (123I-MIBG) is widely used for functional imaging but the added diagnostic value is controversial. OBJECTIVE To establish the virtual impact of adding 123I-MIBG scintigraphy to CT or MRI on diagnosis and treatment of PPGL. DESIGN International multicenter retrospective study. INTERVENTION None. PATIENTS 236 unilateral adrenal, 18 bilateral adrenal, 48 unifocal extra-adrenal, 12 multifocal and 26 metastatic PPGL. MAIN OUTCOME MEASURES Patients underwent both anatomical imaging (CT and/or MRI) and 123I-MIBG scintigraphy. Local imaging reports were analyzed centrally by two independent observers who were blinded to the diagnosis. Imaging-based diagnoses determined by CT/MRI only, 123I-MIBG only, and CT/MRI combined with 123I-MIBG scintigraphy were compared with the correct diagnoses. RESULTS The rates of correct imaging-based diagnoses determined by CT/MRI only versus CT/MRI plus 123I-MIBG scintigraphy were similar: 89.4 versus 88.8%, respectively, (P=0.50). Adding 123I-MIBG scintigraphy to CT/MRI resulted in a correct change in the imaging-based diagnosis and ensuing virtual treatment in four cases (1.2%: two metastatic instead of non-metastatic, one multifocal instead of single, one unilateral instead of bilateral adrenal) at the cost of an incorrect change in seven cases (2.1%: four metastatic instead of non-metastatic, two multifocal instead of unifocal and one bilateral instead of unilateral adrenal). CONCLUSIONS For the initial localization of PPGL, the addition of 123I-MIBG scintigraphy to CT/MRI rarely improves the diagnostic accuracy at the cost of incorrect interpretation in others, even when 123I-MIBG scintigraphy is restricted to patients who are at risk for metastatic disease. In this setting, the impact of 123I-MIBG scintigraphy on clinical decision-making appears very limited