(Young Investigator Award) Trabecular Bone Score Helps Classifying Women At Risk Of Fracture Prospectively In The Ofely Study

Introduction: To determine the metabolic effect of teriparatide (TPTD) on bone, 99mTc-MDP skeletal plasma clearance was measured in postmenopausal women with osteoporosis treated with TPTD 20 μg/day. Methods: Ten postmenopausal women with osteoporosis had radionuclide bone scans at baseline, 3, and 18 months after starting TPTD 20 μg/day and after 6 months off therapy. Participants were injected with 600 MBq 99mTc- MDP and whole body bone scans acquired at 10 min, 1, 2, 3, and 4 h. Multiple blood samples were taken between 5 min and 4 h and free 99mTc-MDP measured using ultrafiltration. 99mTc-MDP plasma clearance (Kbone) was evaluated using the Patlak plot method. Regional differences in Kbone were studied by measuring the whole skeleton and subregions. Serum procollagen type I Nterminal propeptide (PINP), bone-specific alkaline phosphatase (BSAP), and urinary N-terminal telopeptide (NTX) were measured at each visit.Discussion: The median increase from baseline in whole skeleton Kbone was 22% (P=0.004) at 3 months and 34% (P= 0.002) at 18 months, decreasing to 0.7% after 6 months off therapy. In subregions, Kbone value increases were statistically significant at 3 months and in all subregions except the pelvis at 18 months. After 6 months off therapy, subregional Kbone values also returned toward baseline. Bone markers increases from baseline were statistically significant at 3 and 18 months (BSAP, 15% and 36%; PINP, 137% and 192%; NTX, 109% and 125%). After 6 months off therapy, PINP and NTX values had declined, though remained above baseline (BSAP, −3%; PINP, 43%; NTX, 56%). Increased Kbone values in the whole body and lower extremities were correlated with increases in most bone markers at 3 and 18 months. Increased skeletal uptake of 99mTc-MDP during treatment with TPTD is indicative of increased bone formation and is supported by increases in bone turnover markers.Conclusion: Changes in Kbone and skeletal uptake measured by radionuclide bone scans in patients taking TPTD are the result of metabolic activity of the drug. These data may provide physicians with useful insights when interpreting bone scan results in this population.

Two-stage, extreme albitization of A-type granites from Rajasthan, NW India

Albitization is a common process during which hydrothermal fluids convert plagioclase and/or K-feldspar into nearly pure albite; however, its specific mechanism in granitoids is not well understood. The c. 1700 Ma A-type metaluminous ferroan granites in the Khetri complex of Rajasthan, NW India, have been albitized to a large extent by two metasomatic fronts, an initial transformation of oligoclase to nearly pure albite and a subsequent replacement of microcline by albite, with sharp contacts between the microcline-bearing and microcline-free zones. Albitization has bleached the original pinkish grey granite and turned it white. The mineralogical changes include transformation of oligoclase (similar to An(12)) and microcline (similar to Or(95)) to almost pure albite (similar to An(0 center dot 5-2)), amphibole from potassian ferropargasite (X-Fe 0 center dot 84-0 center dot 86) to potassic hastingsite (X-Fe 0 center dot 88-0 center dot 97) and actinolite (X-Fe 0 center dot 32-0 center dot 67), and biotite from annite (X-Fe 0 center dot 71-0 center dot 74) to annite (X-Fe 0 center dot 90-0 center dot 91). Whole-rock isocon diagrams show that, during albitization, the granites experienced major hydration, slight gain in Si and major gain in Na, whereas K, Mg, Fe and Ca were lost along with Rb, Ba, Sr, Zn, light rare earth elements and U. Whole-rock Sm-Nd isotope data plot on an apparent isochron of 1419 +/- 98 Ma and reveal significant disturbance and at least partial resetting of the intrusion age. Severe scatter in the whole-rock Rb-Sr isochron plot reflects the extreme Rb loss in the completely albitized samples, effectively freezing Sr-87/Sr-86 ratios in the albite granites at very high values (0 center dot 725-0 center dot 735). This indicates either infiltration of highly radiogenic Sr from the country rock or, more likely, radiogenic ingrowth during a considerable time lag (estimated to be at least 300 Myr) between original intrusion and albitization. The albitization took place at similar to 350-400 degrees C. It was caused by the infiltration of an ascending hydrothermal fluid that had acquired high Na/K and Na/Ca ratios during migration through metamorphic rocks at even lower temperatures in the periphery of the plutons. Oxygen isotope ratios increase from delta O-18 = 7 parts per thousand in the original granite to values of 9-10 parts per thousand in completely albitized samples, suggesting that the fluid had equilibrated with surrounding metamorphosed crust. A metasomatic model, using chromatographic theory of fluid infiltration, explains the process for generating the observed zonation in terms of a leading metasomatic front where oligoclase of the original granite is converted to albite, and a second, trailing front where microcline is also converted to albite. The temperature gradients driving the fluid infiltration may have been produced by the high heat production of the granites themselves. The confinement of the albitized granites along the NE-SW-trending Khetri lineament and the pervasive nature of the albitization suggest that the albitizing fluids possibly originated during reactivation of the lineament. More generally, steady-state temperature gradients induced by the high internal heat production of A-type granites may provide the driving force for similar metasomatic and ore-forming processes in other highly enriched granitoid bodies.

Harnessing epithelial stem cell potency

Increase in potency of adult stem/progenitor cells holds great expectations for regenerative medicine; reprogramming is achieved by manipulating the genome or indirectly by manipulating the microenvironment. However, the genetic approach, which can result in lineage conversion up to ground pluripotent embryonic state, will certainly face strict regulatory constraints and consequently translation to the clinic may be difficult. Manipulating stem cell fate without altering the genome of adult stem cells is a promising alternative. My laboratory has demonstrated that non hairy squamous epithelia e.g. the cornea, the oral cavity, the oesophagus, the vagina, contain clonogenic stem cells that can respond to skin morphogenetic signals and form epidermis, cycling hair follicles and sebaceous glands. This capacity is maintained in serial transplantation, crosses primary germ line boundaries and is intrinsic to the stem cells, as cells which have never been exposed to cell culture behave in a similar fashion. Even more surprising, the thymus contains a population of clonogenic epithelial cells of endodermal origin that maintain a thymic identity in culture and have the capacity to incorporate into a thymic network, but can acquire the functionality of bona fide multipotent stem cells of the skin when exposed to proper developmental signals. Thymic epithelial cells exposed to a skin microenvironment exhibit a down-regulation or silencing of transcription factors important for thymic function. Hence, it is possible to reveal unsuspected potency and even to robustly reprogram stem cells by solely manipulating the microenvironment.

Non-invasive diagnostic biomarkers for estimating the onset time of permanent cerebral ischemia.

The treatments for ischemic stroke can only be administered in a narrow time-window. However, the ischemia onset time is unknown in ~30% of stroke patients (wake-up strokes). The objective of this study was to determine whether MR spectra of ischemic brains might allow the precise estimation of cerebral ischemia onset time. We modeled ischemic stroke in male ICR-CD1 mice using a permanent middle cerebral artery filament occlusion model with laser Doppler control of the regional cerebral blood flow. Mice were then subjected to repeated MRS measurements of ipsilateral striatum at 14.1 T. A striking initial increase in γ-aminobutyric acid (GABA) and no increase in glutamine were observed. A steady decline was observed for taurine (Tau), N-acetyl-aspartate (NAA) and similarly for the sum of NAA+Tau+glutamate that mimicked an exponential function. The estimation of the time of onset of permanent ischemia within 6 hours in a blinded experiment with mice showed an accuracy of 33±10 minutes. A plot of GABA, Tau, and neuronal marker concentrations against the ratio of acetate/NAA allowed precise separation of mice whose ischemia onset lay within arbitrarily chosen time-windows. We conclude that (1)H-MRS has the potential to detect the clinically relevant time of onset of ischemic stroke.

The alkaline intraplate volcanism of the Antalya nappes (Turkey): a Late Triassic remnant of the Neotethys

Late Triassic submarine alkali basalts and hawaiites were collected from two superimposed tectonic slices belonging to the Kara Dere - Sayrun unit of the Middle Antalya nappes, southwestern Turkey. New determinations on conodont faunas allow to date this sequence to the Lower Carnian (Julian). The volcanic rocks show rather homogeneous compositions, with high TiO2 and relatively low MgO and Ni contents which suggest olivine fractionation. Their primitive mantle-normalised multi-elements plots show Nb and Ta enrichments relative to La, Pb negative anomalies and heavy rare earth element and Y depletions typical of intraplate ocean island basalts. These characteristics are consistent with the major and trace element compositions of their primary clinopyroxene phenocrysts, which do not show any feature ascribable to crustal contamination. The studied lavas display a restricted range of epsilon Nd (+4.6 to +5.2) which falls within the range of ocean island basalts. Their initial (Nd-143/Nd-144)i ratios are too low to be explained by a simple mixing line between depleted MORB mantle (DMM) and HIMU components. Their Pb and Nd isotopic compositions plot along a mixing line between HIMU component and an enriched mantle, the composition of which could be the result of the addition of about 5 to 8% of an EM2 component (recycled marine sediments) to DMM. The lack of evidence for any continental crustal component. in their genesis could be consistent with their emplacement in an intra-oceanic setting.

Shield morphology of curly overhair in 22 genera of Soricidae (Insectivora, Mammalia)

The structure of the shield of curly overhairs was studied in 22 genera of shrews, using scanning electron microscopy. The cross section of the shield is quadriconcave with two sides showing particular scale patterns or a relief which can be grouped into 4 morphological types: 1) a smooth type with, at most, shallow U-shaped notches; 2) a type with uniserial, V-shaped tiled notches; 3) a type with a groove and irregular notches; 4) a type with a deep ridged groove. Myosorex, which occupies a basal phyletic position, shows type 3. This type is therefore interpreted as an ancestral character state. It is found also in Feroculus and in some Sylvisorex. In the Crocidurinae, this type evolved into type 2 (Scutisorex and some Indomalayan Crocidura), and finally into type 1 (Suncus and most Crocidura). In the Soricinae, it evolved into type 4, which is common to all genera except Megasorex and Notiosorex. These two genera possess type 1 which is interpreted as being a synapomorphous character of these genera. The regression of the complex structure under dry climatic conditions supports the hypothesis that the function of the grooved form of hair is water repulsion.

Microenvironmental reprogramming of thymic epithelial cells to skin multipotent stem cells.

The thymus develops from the third pharyngeal pouch of the anterior gut and provides the necessary environment for thymopoiesis (the process by which thymocytes differentiate into mature T lymphocytes) and the establishment and maintenance of self-tolerance. It contains thymic epithelial cells (TECs) that form a complex three-dimensional network organized in cortical and medullary compartments, the organization of which is notably different from simple or stratified epithelia. TECs have an essential role in the generation of self-tolerant thymocytes through expression of the autoimmune regulator Aire, but the mechanisms involved in the specification and maintenance of TECs remain unclear. Despite the different embryological origins of thymus and skin (endodermal and ectodermal, respectively), some cells of the thymic medulla express stratified-epithelium markers, interpreted as promiscuous gene expression. Here we show that the thymus of the rat contains a population of clonogenic TECs that can be extensively cultured while conserving the capacity to integrate in a thymic epithelial network and to express major histocompatibility complex class II (MHC II) molecules and Aire. These cells can irreversibly adopt the fate of hair follicle multipotent stem cells when exposed to an inductive skin microenvironment; this change in fate is correlated with robust changes in gene expression. Hence, microenvironmental cues are sufficient here to re-direct epithelial cell fate, allowing crossing of primitive germ layer boundaries and an increase in potency.

Anti-idiotypic monoclonal antibody AB3, reacting with the primary antigen (CEA), can localize in human colon-carcinoma xenografts as efficiently as AB1.

BALB/c mice were immunized with anti-idiotypic monoclonal (MAb) antibody (anti-Id or Ab2) directed against an AB1 MAb anti-carcinoembryonic (CEA) in order to obtain AB3 MAbs (anti-anti-Id). AB3 MAbs were shown to recognise the primary antigen (CEA) and one of them was tested extensively in vitro and in vivo. This AB3 MAb was shown to bind specifically to CEA on frozen sections of a human colon carcinoma by immunoperoxidase. Scatchard plot analyses showed that the affinity of this AB3 was of the same order of magnitude as the AB1. In vivo experiments, in nude mice bearing CEA-producing human colon-carcinoma xenografts showed that up to 30% of the intravenously injected dose of 125I-labelled AB3 were localized per gram of tumour tissue. Furthermore, calculation of the ratios of AB3 concentration in the tumour over those in normal organs such as lung, liver, kidney, spleen and bone gave relatively high values similar to results obtained with AB1. All together our results show that AB3 can localize as efficiently and specifically in the tumour as AB1, despite the fact that the mice from which it was derived were immunized with a mouse MAb (AB2) and had never been exposed to CEA.

Role of Notch signaling in epidermis and appendages

Résumé Dans la peau, il a été montré que Notch1 induit l'arrêt de la prolifération et la différentiation des keratinocytes. L'inactivation de Notch1 cause une hyperplasie de l'épiderme et la formation de carcinomes basaux cellulaires. Notre groupe a principalement identifié deux voies de signalisations, la voie Shh et la voie Wnt, qui sont dérégulées en conséquence de l'inactivation de Notch1 dans la peau. Nous avons démontré l'habilité de Notch1 à réprimer la voie Wnt induite par ß-catenin dans les keratinocytes primaires ainsi que dans d'autres types de cellules épithéliales humaines. De plus, nous avons pu déterminer que Notch1 régule cette voie, probablement en favorisant la phosphorylation de ß-catenin par le complexe axin/APC/GSK-3ß. La protéine faisant partie de la voie Wnt, ou la protéine affectant la voie Wnt, qui est régulée par Notch1 est sujette à de plus amples investigations. Un autre but de cette étude a été l'identification de potentiels gènes cibles de Notch1 autres que ceux faisant partie des voies de signalisation Shh et Wnt précédemment évoquées. Ce projet fut abordé par l'analyse de puces à ADN (ISREC et Affymetrix) qui ont été utilisées pour des expériences de gain et de perte de fonction de Notch1 dans des keratinocytes prúmaires. En plus de l'hyperplasie épidermale, les souris Notch1 déficiente ont une perte importante de poils. Nous avons montré que Notch1 est nécessaire pour le développement et l'homéostasie des follicules pileux. En effet, l'inactivation du gène Notch1 mediée par l'activation des kératines 5 ou 14 dans l'épiderme, cause des défauts du cycle ainsi que de la structure des poils. De plus, d'autres appendices de la peau, comme les glandes sudoripares et de Meibomius, ont une structure anormale et sont non fonctionnelles dans les souris Notch1 déficiente. Finalement, nous avons observé que la déficience de Notch1 dans l'épithélium cornéen mène à la formation d'une plaque épidermale opaque sur la cornée. Basé sur l'hypothèse que le défaut des glandes de Meibomius des souris Notch1 déficientes cause des lésions de la surface oculaire, nous avons montré que Notch1 est essentiel pour la cicatrisation de la cornée. Lorsque Notch1 est absent, les cellules souches de l'épithélium cornéen ne sont plus capables de se différentier en cellules cornéennes, mais réparent la blessure en se différentiant en épiderme. Ce résultat indique que Notch1 est essentiel pour la différentiation de cellules souches de la cornée qui sont spécifiquement impliquées dans la réparation de la cornée. De plus, nous avons montré que l'expression de CRBP1 dans l'épithélium cornéen est diminuée en l'absence de Notch1, ceci étant possiblement à l'origine de la formation de la plaque épidermale. Abstract: In the skin, Notch1 has been shown to trigger cell growth arrest and differentiation of keratinocytes. Notch1 inactivation results in epidermal hyperplasia and subsequent formation of basal cell carcinoma-like (BCC-like) tumors. So far our group has identified two main pathways, the Shh and the Wnt pathway, that are deregulated as a consequence of Notch1 inactivation in the skin. We showed the ability of Notch1 to represses ß-catenin-mediated Wnt signaling in primary keratinocytes as well as in other types of human epithelial cells. In addition we were able to determine that Notch1 regulates this pathway possibly by enhancing ß-catenin phosphorylation by the axin/APC/GSK-3ß complex. The exact target protein of the Wnt pathway or target protein that affects the Wnt pathway, and that is regulated by Notch1, is subject of current investigation. Another aim of this study was the identification of possible Notch1 target genes in addition to those of the Shh and Wnt signaling pathways. This was addressed by gene chip analysis using ISREC as well as Affymetrix microarrays for gain and loss of function of Notch1 in mouse primary keratinocytes. In addition to epidermal hyperplasia, Notch1 deficient mice show an important hair loss. We showed that Notch1 is required for postnatal development and homeostasis of hair follicles. Indeed, keratin5 or keratinl4-driven Cre recombinase-mediated inactivation of the Notch1 gene in the epidermis causes perturbations of the hair cycle and structural defects of the hair follicle. Moreover, other skin appendages, like the sweat and Meibomian glands show abnormal morphology and are not functional in the Notch 1 deficient mice. Finally, we observed that Notch1 deficiency in the corneal epithelium leads to the formation of an epidermal corneal plaque. Based on the hypothesis that the Meiboinian gland defect in the Notch1 deficient mice results in lesions of the eye surface, we showed that Notch1 is essential for wound-healing of the cornea. In absence of Notch1 the stem cells of the corneal epithelium are no longer able to differentiate in the corneal fate but instead repair the wound by differentiating into skin-like epidermis. This result indicated that Notch1 is essential for the differentiation of corneal stem cells specifically implicated in corneal wound-healing. Moreover, we showed that CRBP1 expression in the corneal epithelium was lost in the absence of Notch1, possibly being at the origin of plaque formation.

Three-dimensional ordered-subset expectation maximization iterative protocol for evaluation of left ventricular volumes and function by quantitative gated SPECT: a dynamic phantom study.

The purposes of this study were to characterize the performance of a 3-dimensional (3D) ordered-subset expectation maximization (OSEM) algorithm in the quantification of left ventricular (LV) function with (99m)Tc-labeled agent gated SPECT (G-SPECT), the QGS program, and a beating-heart phantom and to optimize the reconstruction parameters for clinical applications. METHODS: A G-SPECT image of a dynamic heart phantom simulating the beating left ventricle was acquired. The exact volumes of the phantom were known and were as follows: end-diastolic volume (EDV) of 112 mL, end-systolic volume (ESV) of 37 mL, and stroke volume (SV) of 75 mL; these volumes produced an LV ejection fraction (LVEF) of 67%. Tomographic reconstructions were obtained after 10-20 iterations (I) with 4, 8, and 16 subsets (S) at full width at half maximum (FWHM) gaussian postprocessing filter cutoff values of 8-15 mm. The QGS program was used for quantitative measurements. RESULTS: Measured values ranged from 72 to 92 mL for EDV, from 18 to 32 mL for ESV, and from 54 to 63 mL for SV, and the calculated LVEF ranged from 65% to 76%. Overall, the combination of 10 I, 8 S, and a cutoff filter value of 10 mm produced the most accurate results. The plot of the measures with respect to the expectation maximization-equivalent iterations (I x S product) revealed a bell-shaped curve for the LV volumes and a reverse distribution for the LVEF, with the best results in the intermediate range. In particular, FWHM cutoff values exceeding 10 mm affected the estimation of the LV volumes. CONCLUSION: The QGS program is able to correctly calculate the LVEF when used in association with an optimized 3D OSEM algorithm (8 S, 10 I, and FWHM of 10 mm) but underestimates the LV volumes. However, various combinations of technical parameters, including a limited range of I and S (80-160 expectation maximization-equivalent iterations) and low cutoff values (< or =10 mm) for the gaussian postprocessing filter, produced results with similar accuracies and without clinically relevant differences in the LV volumes and the estimated LVEF.

Peroxisome proliferator-activated receptor (PPAR)-beta as a target for wound healing drugs: what is possible?

Peroxisome proliferator-activated receptor (PPAR) dysfunction has been implicated in the manifestation of many diseases and illnesses, ranging from obesity to cancer. Herein, we discuss the role of PPARbeta, one of the three PPAR isotypes, during wound healing. While PPARbeta expression is undetectable in unchallenged and healthy adult interfollicular mouse skin, it is robustly re-activated in stress situations, such as upon phorbol ester treatment, hair plucking and cutaneous wounding. The inflammatory reaction associated with a skin injury activates the keratinocytes at the edges of the wound. This activation involves PPARbeta, whose expression and activity as transcription factor are up-regulated by pro-inflammatory signals. The re-activation of PPARbeta influences three important properties of the activated keratinocytes that are vital for rapid wound closure, namely, survival, migration and differentiation. The anti-apoptotic and, thus, survival role of PPARbeta is mediated by the up-regulation of expression of integrin-linked kinase and 3-phosphoinositide-dependent kinase-1. Both kinases are required for the full activation of the Akt1 survival cascade. Therefore, the up-regulation of PPARbeta, early after injury, appears to be important to maintain a sufficient number of viable keratinocytes at the wound edge. At a later stage of wound repair, the stimulation of keratinocyte migration and differentiation by PPARbeta is also likely to be important for the formation of a new epidermis at the wounded area. Consistent with these observations, the entire wound healing process is delayed in PPARbeta +/- mice and wound closure is retarded by 2-3 days. The multiple roles of PPARbeta in the complex keratinocyte response after injury and during skin repair certainly justify a further exploration of its potential as a target for wound healing drugs.

Selective expression of a dominant-negative form of peroxisome proliferator-activated receptor in keratinocytes leads to impaired epidermal healing.

Many nuclear hormone receptors are involved in the regulation of skin homeostasis. However, their role in the epithelial compartment of the skin in stress situations, such as skin healing, has not been addressed yet. The healing of a skin wound after an injury involves three major cell types: immune cells, which are recruited to the wound bed; dermal fibroblasts; and epidermal and hair follicle keratinocytes. Our previous studies have revealed important but nonredundant roles of PPARalpha and beta/delta in the reparation of the skin after a mechanical injury in the adult mouse. However, the mesenchymal or epithelial cellular compartment in which PPARalpha and beta/delta play a role could not be determined in the null mice used, which have a germ line PPAR gene invalidation. In the present work, the role of PPARalpha was studied in keratinocytes, using transgenic mice that express a PPARalpha mutant with dominant-negative (dn) activity specifically in keratinocytes. This dn PPARalpha lacks the last 13 C terminus amino acids, binds to a PPARalpha agonist, but is unable to release the nuclear receptor corepressor and to recruit the coactivator p300. When selectively expressed in keratinocytes of transgenic mice, dn PPARalphaDelta13 causes a delay in the healing of skin wounds, accompanied by an exacerbated inflammation. This phenotype, which is similar to that observed in PPARalpha null mice, strongly suggests that during skin healing, PPARalpha is required in keratinocytes rather than in other cell types.

Loss of Memo, a novel FGFR regulator, results in reduced lifespan.

Memo is a widely expressed 33-kDa protein required for heregulin (HRG)-, epidermal growth factor (EGF)-, and fibroblast growth factor (FGF)-induced cell motility. Studies in mouse embryonic fibroblasts, wild-type or knockout for Memo, were performed to further investigate the role of Memo downstream of FGFR. We demonstrated that Memo associates with the FGFR signalosome and is necessary for optimal activation of signaling. To uncover Memo's physiological role, Memo conditional-knockout mice were generated. These animals showed a reduced life span, increased insulin sensitivity, small stature, graying hair, alopecia, kyphosis, loss of subcutaneous fat, and loss of spermatozoa in the epididymis. Memo-knockout mice also have elevated serum levels of active vitamin D, 1,25-dihydroxyvitamin D3 (1,25(OH)2D), and calcium compared to control littermates expressing Memo. In summary, the results from in vivo and in vitro models support the hypothesis that Memo is a novel regulator of FGFR signaling with a role in controlling 1,25(OH)2D production and normal calcium homeostasis.

Ethyl glucuronide in plant extracts :O16

Introduction: The specificity of ethyl glucuronide (EtG) in hair as marker of alcohol consumption exceeds by far those of fatty acid ethyl esters. False positive cases are therefore very rare but not excluded as recent publications have shown. Especially, the use of plant extracts containing high percentages of ethanol can lead to EtG hair concentrations typically found in cases of chronic alcohol consumption. As proposed by Baumgartner et al., a nucleohilic substitution could most likely explain this phenomenon. Fresh and dried plants as well as commercial hair lotions based on plants extracts have been analysed for EtG presence or EtG formation. Methods: Urtica dioica, Plantago lanceolata, Cortex Quercus, Sempervivum, Armoracia rusticana, Juniperus communis, Brassica alba, Thymian vulgaris, Salvia officinalis, Majorana hortensis, Aloe vera, birch gingko and green tea leafs, ginger, lemon grass were extracted in water, water/ethanol (50/50) and ethanol (100%). The extracts as well as diluted hair lotions were measured by immunological test (Microgenics DRI® EtG assay) and by LC-MS/MS on Shimadzu Nexera UHPLC coupled with an AB Sciex 4500 QTrap. Results: EtG could not be detected in water extracts of all tested plants. However, DRI® EtG assay indicated the presence of EtG in 66% of the tested ethanolic plant extracts. That could only be confirmed by mass spectrometry in the cases of fresh thyme as well as in dried birch, oak and plantain extracts where EtG concentrations between of 0.25 and 2,09 mg/l were measured. In one hair lotion, the EtG concentration was 0,76 mg/l. Conclusion: Ethanolic plant extracts represents a non-negligible risk for false positive EtG hair tests, especially when applied as lotion without following washing out. The use of hair care products must therefore be evaluated at every hair sampling. In case of doubt, the product should be analysed by mass spectrometric methods since the presence of EtG can't be proven by use of the DRI® EtG assay, only. Our results support Baumgartner's assumption of a nucleophilic substitution in presence of ethanol because EtG was only measured in the ethanolic extracts.

Impact of free-range pigs on mountain pastures in the Swiss Jura

The consequences of foraging by free-range pigs on the vegetation of mountain pastures was investigated. 25 pigs (15 week-old, mean weight 50 kg) were enclosed from June to mid-September in a 2 ha-enclosure in Jura Mountains (Switzerland), fed with a mixture of lactoserum and cereals. The enclosure contained five different plant communities. Eutrophic pastures on deep soil were strongly overturned, but the recolonisation was quick and dominated by the original species. Mesotrophic pastures were less damaged on stony soil but completely destroyed on deep soil, and the recovery was slow, characterised by a shift of plant species in a more eutrophic direction. Four years were not sufficient for complete recovery. Oligotrophic calcareous pastures on shallow stony soil were not damaged. Extensive breeding of pigs in mountain pastures might be harmful to plant species and vegetation, and ought to be restricted to the less sensitive plant communities, with a rotation on two to three different sites.