Suction blister fluid as potential body fluid for biomarker proteins

Early diagnosis is important for effective disease management. Measurement of biomarkers present at the local level of the skin could be advantageous in facilitating the diagnostic process. The analysis of the proteome of suction blister fluid, representative for the interstitial fluid of the skin, is therefore a desirable first step in the search for potential biomarkers involved in biological pathways of particular diseases. Here, we describe a global analysis of the suction blister fluid proteome as potential body fluid for biomarker proteins. The suction blister fluid proteome was compared with a serum proteome analyzed using identical protocols. By using stringent criteria allowing less than 1% false positive identifications, we were able to detect, using identical experimental conditions and amount of starting material, 401 proteins in suction blister fluid and 240 proteins in serum. As a major result of our analysis we construct a prejudiced list of 34 proteins, relatively highly and uniquely detected in suction blister fluid as compared to serum, with established and putative characteristics as biomarkers. We conclude that suction blister fluid might potentially serve as a good alternative biomarker body fluid for diseases that involve the skin.     

Intermediate stage complex regional pain syndrome type 1 is unrelated to proinflammatory cytokines

The aim of this paper is to determine the involvement of tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-6 in intermediate CRPS 1 as locally formed mediators of inflammation. In this study, 25 patients with proven CRPS 1 (Bruehl criteria) were included. All patients participated in one of our earlier studies during the acute stage of their disease. After the disease developed into an intermediate stage, both the disease activity and the profile of inflammatory mediators were reevaluated. Disease activity and impairment were determined by means of a visual analogue scale, the McGill Pain Questionnaire, the difference in volume and temperature between the involved and uninvolved extremities, and the reduction in active range of motion of the involved extremity. Suction blisters were made on the involved and uninvolved extremities for measurement of IL-6 and TNF-alpha. A significant improvement in signs and symptoms of impairment was found. However, the levels of IL-6 and TNF-alpha in blister fluid in the involved extremity versus uninvolved extremity were still significantly raised. Although signs and symptoms are significantly improved, proinflammatory cytokines are still increased in CRPS 1 affected extremities during the intermediate stage of the disease. This indicates that the initiation and sustained development of the disease are only partially affected by proinflammatory cytokines. Follow-up in the chronic stage is necessary to draw more definite conclusions about the existence of a supposed relation between clinical signs and symptoms and the level of proinflammatory cytokines.     

Charred olive stones: experimental and archaeological evidence for recognizing olive processing residues used as fuel

After extracting oil from olives a residue is left usually referred to as the olive oil processing residue (OPR). This study explores the way in which ancient societies may have used OPR as fuel for fires to generate heat and the various issues that are related to the residues of this fuel. After drying, the high heating value and structure of OPR makes it an excellent and efficient fuel. Upgrading OPR further, through thermal conversion or charring, provides an even more efficient fuel (COPR), with a hotter and smoke free flame, a higher heating value and which is lighter in mass and thus easier to transport. After a fire is extinguished two types of remains of the fuel are left i.e. char and ash. Analyses on both remains, recovered from archaeological deposits, could be used as a source of information on fuel utilization. Laboratory experiments on charred modern OPR and stones show that by measuring their reflectance and analyzing their structure under reflected light microscopy, OPR and COPR can be distinguished in the charred material recovered from three archaeological sites in Greece and Syria. Based on these investigations it is suggested that on the three sites COPR was used as fuel. Ash, sampled together with the char, provides the possibility of investigating if other types of fuel were used, apart from OPR or COPR. On the investigated sites no ash was collected, but the analysis of the modern OPR showed that the properties of its ash could be used to distinguish it from other types of fuel. Ash from modern OPR and olive stones showed the presence of phytoliths. The often discussed issue related to the sharpness and smoothness of the edges of charred fragmented olive stones was investigated. The results showed that this is not a reliable criterion for recognizing olive oil production. It is recommended that in addition to the identification of the botanical material more properties of the remains of fuels should be analysed. To prevent destroying and losing char and ash as a result of excavation activities such as flotation and sieving, special measures have to be taken. The results show that analysing char and ash may provide valuable information on the (pyro)technology practised in ancient societies.     

Proximal Spectral Sensing to Monitor Phytoremediation of Metal-Contaminated Soils

Assessment of soil contamination and its long-term monitoring are necessary to evaluate the effectiveness of phytoremediation systems. Spectral sensing-based monitoring methods promise obvious benefits compared to field-based methods: lower cost, faster data acquisition and better spatio-temporal monitoring. This paper reviews the theoretical basis whereby proximal spectral sensing of soil and vegetation could be used to monitor phytoremediation of metal-contaminated soils, and the eventual upscaling to imaging sensing. Both laboratory and field spectroscopy have been applied to sense heavy metals in soils indirectly via their intercorrelations with soil constituents, and also through metal-induced vegetation stress. In soil, most predictions are based on intercorrelations of metals with spectrally-active soil constituents viz., Fe-oxides, organic carbon, and clays. Spectral variations in metal-stressed plants is particularly associated with changes in chlorophyll, other pigments, and cell structure, all of which can be investigated by vegetation indices and red edge position shifts. Key shortcomings in obtaining satisfactory calibration for monitoring the metals in soils or metal-related plant stress include: reduced prediction accuracy compared to chemical methods, complexity of spectra, no unique spectral features associated with metal-related plant stresses, and transfer of calibrations from laboratory to field to regional scale. Nonetheless, spectral sensing promises to be a time saving, non-destructive and cost-effective option for long-term monitoring especially over large phytoremediation areas, and it is well-suited to phytoremediation networks where monitoring is an integral part.     

LEAF SHAPE EVOLUTION IN THE SOUTH AFRICAN GENUS PELARGONIUM L' HÉR. (GERANIACEAE)

Leaf shapes reflect complex assemblages of shape-determining elements, yet evolutionary studies tend to treat leaf shape as a single attribute, for example cordate or linear. As with all complex structures, individual elements of a leaf could theoretically evolve independently and at different rates to the extent permitted by genetic and functional limitations. We examined relative evolutionary lability of shape-determining elements in the highly diverse South African plant genus Pelargonium (Geraniaceae). We used SIMMAP to calculate Bayesian posterior probabilities for ancestral states of leaf-shape characters for major nodes across multiple phylogenetic trees. Trees were derived from a Bayesian analysis of DNA sequence data from four partitions. We found that shape elements differed in rates of character-state transformations across the tree. Leaf base, apex, and overall outline had low rates. Transformations in venation occurred at slightly higher rates and were associated with shifts in venation among major clades. Leaf margin type and overall leaf size showed intermediate rates, whereas high rates were observed in the extent of lamina lobing and functional leaf size. The results indicate that suites of elements characteristic of the recently evolved xerophytic lineage, for example pinnate venation, dissected lamina, and entire margins, were acquired piecemeal over nested levels of the phylogeny.     

Olfactory receptor genes cooperate with protocadherin genes in human extreme obesity

Worldwide, the incidence of obesity has increased dramatically over the past decades. More knowledge about the complex etiology of obesity is needed in order to find additional approaches for treatment and prevention. Investigating the exome sequencing data of 30 extremely obese subjects (BMI 45–65 kg/m²) shows that predicted damaging missense variants in olfactory receptor genes on chromosome 1q and rare predicted damaging variants in the protocadherin (PCDH) beta-cluster genes on chromosome 5q31, reported in our previous work, co-localize in subjects with extreme obesity. This implies a synergistic effect between genetic variation in these gene clusters in the predisposition to extreme obesity. Evidence for a general involvement of the olfactory transduction pathway on itself could not be found. Bioinformatic analysis indicates a specific involvement of the PCDH beta-cluster genes in controlling tissue development. Further mechanistic insight needs to await the identification of the ligands of the 1q olfactory receptors. Eventually, this may provide the possibility to manipulate food flavor in a way to reduce the risk of overeating and of extreme obesity in genetically predisposed subjects.

Electronic supplementary material

The online version of this article (doi:10.1007/s12263-015-0465-3) contains supplementary material, which is available to authorized users.     

Targeted mutation of the talpid3 gene in zebrafish reveals its conserved requirement for ciliogenesis and Hedgehog signalling across the vertebrates

Using zinc-finger nuclease-mediated mutagenesis, we have generated mutant alleles of the zebrafish orthologue of the chicken talpid3 (ta3) gene, which encodes a centrosomal protein that is essential for ciliogenesis. Animals homozygous for these mutant alleles complete embryogenesis normally, but manifest a cystic kidney phenotype during the early larval stages and die within a month of hatching. Elimination of maternally derived Ta3 activity by germline replacement resulted in embryonic lethality of ta3 homozygotes. The phenotype of such maternal and zygotic (MZta3) mutant zebrafish showed strong similarities to that of chick ta3 mutants: absence of primary and motile cilia as well as aberrant Hedgehog (Hh) signalling, the latter manifest by the expanded domains of engrailed and ptc1 expression in the somites, reduction of nkx2.2 expression in the neural tube, symmetric pectoral fins, cyclopic eyes and an ectopic lens. GFP-tagged Gli2a localised to the basal bodies in the absence of the primary cilia and western blot analysis showed that Gli2a protein is aberrantly processed in MZta3 embryos. Zygotic expression of ta3 largely rescued the effects of maternal depletion, but the motile cilia of Kupffer's vesicle remained aberrant, resulting in laterality defects. Our findings underline the importance of the primary cilium for Hh signaling in zebrafish and reveal the conservation of Ta3 function during vertebrate evolution.     

The Microbial Diversity of Traditional Spontaneously Fermented Lambic Beer

Lambic sour beers are the products of a spontaneous fermentation that lasts for one to three years before bottling. The present study determined the microbiota involved in the fermentation of lambic beers by sampling two fermentation batches during two years in the most traditional lambic brewery of Belgium, using culture-dependent and culture-independent methods. From 14 samples per fermentation, over 2000 bacterial and yeast isolates were obtained and identified. Although minor variations in the microbiota between casks and batches and a considerable species diversity were found, a characteristic microbial succession was identified. This succession started with a dominance of Enterobacteriaceae in the first month, which were replaced at 2 months by Pediococcus damnosus and Saccharomyces spp., the latter being replaced by Dekkera bruxellensis at 6 months fermentation duration.