Genome-wide association study identifies novel loci associated with circulating phospho- and sphingolipid concentrations

Phospho- and sphingolipids are crucial cellular and intracellular compounds. These lipids are required for active transport, a number of enzymatic processes, membrane formation, and cell signalling. Disruption of their metabolism leads to several diseases, with diverse neurological, psychiatric, and metabolic consequences. A large number of phospholipid and sphingolipid species can be detected and measured in human plasma. We conducted a meta-analysis of five European family-based genome-wide association studies (N = 4034) on plasma levels of 24 sphingomyelins (SPM), 9 ceramides (CER), 57 phosphatidylcholines (PC), 20 lysophosphatidylcholines (LPC), 27 phosphatidylethanolamines (PE), and 16 PE-based plasmalogens (PLPE), as well as their proportions in each major class. This effort yielded 25 genome-wide significant loci for phospholipids (smallest P-value = 9.88×10⁻²⁰⁴) and 10 loci for sphingolipids (smallest P-value = 3.10×10⁻⁵⁷). After a correction for multiple comparisons (P-value<2.2×10⁻⁹), we observed four novel loci significantly associated with phospholipids (PAQR9, AGPAT1, PKD2L1, PDXDC1) and two with sphingolipids (PLD2 and APOE) explaining up to 3.1% of the variance. Further analysis of the top findings with respect to within class molar proportions uncovered three additional loci for phospholipids (PNLIPRP2, PCDH20, and ABDH3) suggesting their involvement in either fatty acid elongation/saturation processes or fatty acid specific turnover mechanisms. Among those, 14 loci (KCNH7, AGPAT1, PNLIPRP2, SYT9, FADS1-2-3, DLG2, APOA1, ELOVL2, CDK17, LIPC, PDXDC1, PLD2, LASS4, and APOE) mapped into the glycerophospholipid and 12 loci (ILKAP, ITGA9, AGPAT1, FADS1-2-3, APOA1, PCDH20, LIPC, PDXDC1, SGPP1, APOE, LASS4, and PLD2) to the sphingolipid pathways. In large meta-analyses, associations between FADS1-2-3 and carotid intima media thickness, AGPAT1 and type 2 diabetes, and APOA1 and coronary artery disease were observed. In conclusion, our study identified nine novel phospho- and sphingolipid loci, substantially increasing our knowledge of the genetic basis for these traits.     

Towards high throughput metabolic flux analysis in plants

Research on plant metabolism is currently experiencing the common use of various omics methods creating valuable information on the concentrations of the cell's constituents. However, little is known about in vivo reaction rates, which can be determined by Metabolic Flux Analysis (MFA), a combination of isotope labeling experiments and computer modeling of the metabolic network. Large-scale applications of this method so far have been hampered by tedious procedures of tissue culture, analytics, modeling and simulation. By streamlining the workflow of MFA, the throughput of the method could be significantly increased. We propose strategies for these improvements on various sub-steps which will move flux analysis to the medium-throughput range and closer to established methods such as metabolite profiling. Furthermore, this may enable novel applications of MFA, for example screening plant populations for traits related to the flux phenotype.     

Protein p0071, a major plaque protein of non-desmosomal adhering junctions,is a selective cell-type marker

Protein p0071, which originally was introduced as a member of the p120-subfamily of armadillo proteins, common to desmosomes and adhaerens junctions (AJs) and to several other cell structures (centrosomes, midbodies), has been localized by using a series of novel mono- and polyclonal antibodies generated against various domains of the molecule. By protein analysis and immunolocalization techniques, protein p0071 has been localized as a plaque protein in AJs of diverse epithelia and certain vascular endothelia, in the composite junctions (areal compositae) of the intercalated disks of cardiomyocytes, and in the punctate or more extended AJs of the vast majority of cell culture types examined, including mitotic states. Using these antibodies, we have also shown that this AJ protein occurs only rarely or is even absent in tissues such as skeletal and smooth muscles, in a series of mesenchymal tissue cells, and in specific desmosome-rich cells such as those of the upper layers of the epidermis and certain other stratified epithelia and Hassall corpuscles of the thymus. We have also demonstrated that p0071 is absent from desmosomes. The occurrence of two major subtypes of lymphatic endothelial cells, one with AJs containing p0071 and one without detectable p0071, is emphasized. Possible structural and functional roles of p0071 are discussed in light of these new findings regarding its localization, and the addition of p0071 to the armamentarium of cytodiagnostic cell-type markers is recommended. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. This study was supported by project grants from the German Ministry for Education and Research (BMBF) in a cooperative research program entitled “Standardization of mesenchymal stem cells for regenerative medicine (START-MSC)” and from the Deutsche Krebshilfe (project 10–2049) to W.W. Franke.     

Subtypes of melanocytes and melanoma cells distinguished by their intercellular contacts: heterotypic adherens junctions,adhesive associations,and dispersed desmoglein 2 glycoproteins

In the tissue integration of melanocytes and melanoma cells, an important role is attributed to cell adhesion molecules, notably the cadherins. In cultured melanoma cells, we have previously described a more heterogeneous repertoire of cadherins than normal, including some melanoma subtypes synthesizing the desmosomal cadherin, desmoglein 2, out of the desmosomal context. Using biochemical and immunological characterization of junctional molecules, confocal laser scanning, and electron and immunoelectron microscopy, we now demonstrate homo- and heterotypic cell-cell adhesions of normal epidermal melanocytes. In human epidermis, both in situ and in cell culture, melanocytes and keratinocytes are connected by closely aligned membranes that are interspersed by small puncta adhaerentia containing heterotypic complexes of E- and P-cadherin. Moreover, melanocytes growing in culture often begin to synthesize desmoglein 2, which is dispersed over extended areas of intimate adhesive cell-cell associations. As desmoglein 2 is not found in melanocytes in situ, we hypothesize that its synthesis is correlated with cell proliferation. Indeed, in tissue microarrays, desmoglein 2 has been demonstrated in a sizable subset of nevi and primary melanomas. The biological meanings of these cell-cell adhesion molecule arrangements, the possible diagnostic and prognostic significance of these findings, and the implications of the heterogeneity types of melanomas are discussed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. This work was supported in parts by grants from the Deutsche Forschungsgemeinschaft to W. K. Peitsch (project PE 896/1) and the Deutsche Krebshilfe to W. W. Franke (project 10-2049).     

Predicting species distributions in poorly-studied landscapes

Conservationists are increasingly relying on distribution models to predict where species are likely to occur, especially in poorly-surveyed but biodiverse areas. Modeling is challenging in these cases because locality data necessary for model formation are often scarce and spatially imprecise. To identify methods best suited to modeling in these conditions, we compared the success of three algorithms (Maxent, Mahalanobis Typicalities and Random Forests) at predicting distributions of eight bird and eight mammal species endemic to the eastern slopes of the central Andes. We selected study species to have a range of locality sample sizes representative of the data available for endemic species of this region and also that vary in their distribution characteristics. We found that for species that are known from moderate numbers (N = 38–94) of localities, the three methods performed similarly for species with restricted distributions but Maxent and Random Forests yielded better results for species with wider distributions. For species with small numbers of sample localities (N = 5–21), Maxent produced the most consistently successful results, followed by Random Forests and then Mahalanobis Typicalities. Because evaluation statistics for models derived from few localities can be suspect due to the poor spatial representation of the evaluation data, we corroborated these results with review by scientists familiar with the species in the field. Overall, Maxent appears to be the most capable method for modeling distributions of Andean bird and mammal species because of the consistency of results in varying conditions, although the other methods have strengths in certain situations. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Age- and fitness-related differences in limb venous compliance do not affect tolerance to maximal lower body negative pressure in men and women.

Aging and chronic exercise training influence leg venous compliance. Venous compliance affects responses to an orthostatic stress; its effect on tolerance to maximal lower body negative pressure (LBNP) in the elderly is unknown. The purpose of this study was to determine the influence of age and fitness, a surrogate measure of exercise training, on calf venous compliance and tolerance to maximal LBNP in men and women. Forty participants, 10 young fit (YF; age = 22.6 +/- 0.5 yr, peak oxygen uptake = 57.1 +/- 2.0 ml.kg(-1).min(-1)), 10 young unfit (YU; 23.1 +/- 1.0 yr, 41.1 +/- 2.0 ml.kg(-1).min(-1)), 10 older fit (OF; 73.9 +/- 2.0 yr, 39.0 +/- 2.0 ml.kg(-1).min(-1)), and 10 older unfit (OU; 70.9 +/- 1.6 yr, 27.1 +/- 2.0 ml.kg(-1).min(-1)), underwent graded LBNP to presyncope or 4 min at -100 mmHg. By utilizing venous occlusion plethysmography, calf venous compliance was determined by using the first derivative of the pressure-volume relation during cuff pressure reduction. We found that the more fit groups had greater venous compliance than their unfit peers (P < 0.05) as did the young groups compared with their older peers (P < 0.05) such that OU < YU = OF < YF. LBNP tolerance did not differ between groups. In conclusion, these data suggest that aging reduces, and chronic exercise increases, venous compliance. However, these data do not support a significant influence of venous compliance on LBNP tolerance.     

Folic acid deficiency anemia caused by therapy in chronic renal insufficiency

The turbidimetric determination of folic acid concentration in serum and erythrocytes was made by means of a spectrophotometer multiscan in 35 conservatively treated patients with chronic renal failure with a minimum creatine retention of 309 mumol/l and in 63 hemodialysed patients with terminal renal insufficiency. The result showed a statistically significant diminution of serum folic acid concentration in those patients treated in a conservative manner and a significant diminution of plasmatic and erythrocytic folate++ activity in patients under hemodialysis. There was a correlation between the folic acid concentration and the hemoglobin level in both groups of patients, whereas there was no correlation to the number of leukocytes and thrombocytes. A protein deficiency diet or loss of dialysis through the capillary membrane could be found to be the cause for the loss of folic acid. The possibility of folate++ substitutive therapy and its performance was discussed and recommended for selected indications.     

Structure-activity relationships in polycyclic aromatic hydrocarbons: induction of microsomal aryl hydrocarbon hydroxylase and its possible importance in chemical carcinogenesis

The results of quantitative structure-activity analysis show that the potency of polycyclic hydrocarbons as inducers of microsomal arylhydrocarbon hydroxylase (AHH) can be quantitatively described in terms of hydrophobic interactions, chemical reactivity, and the ability to participate in charge-transfer interactions. The rate-determining step in enzyme induction is probably the formation of a reactive K-region metabolite of the hydrocarbons which then reacts with a specific repressor. The same mechanism is proposed for chemical carcinogenesis on the ground of similarities in structure-activity relationships for both processes. It is suggested that AHH induction is the first step in chemical carcinogenesis.