High molecular diversity in the true service tree (Sorbus domestica) despite rareness: data from Europe with special reference to the Austrian occurrence

Background and Aims Sorbus domestica (Rosaceae) is one of the rarest deciduous tree species in Europe and is characterized by a scattered distribution. To date, no large-scale geographic studies on population genetics have been carried out. Therefore, the aims of this study were to infer levels of molecular diversity across the major part of the European distribution of S. domestica and to determine its population differentiation and structure. In addition, spatial genetic structure was examined together with the patterns of historic and recent gene flow between two adjacent populations.Methods Leaf or cambium samples were collected from 17 populations covering major parts of the European native range from north-west France to south-east Bulgaria. Seven nuclear microsatellites and one chloroplast minisatellite were examined and analysed using a variety of methods.Key Results Allelic richness was unexpectedly high for both markers within populations (mean per locus: 3·868 for nSSR and 1·647 for chloroplast minisatellite). Moreover, there was no evidence of inbreeding (mean F_is = –0·047). The Italian Peninsula was characterized as a geographic region with comparatively high genetic diversity for both genomes. Overall population differentiation was moderate (F_ST = 0·138) and it was clear that populations formed three groups in Europe, namely France, Mediterranean/Balkan and Austria. Historic gene flow between two local Austrian populations was high and asymmetric, while recent gene flow seemed to be disrupted.Conclusions It is concluded that molecular mechanisms such as self-incompatibility and high gene flow distances are responsible for the observed level of allelic richness as well as for population differentiation. However, human influence could have contributed to the present genetic pattern, especially in the Mediterranean region. Comparison of historic and recent gene flow may mirror the progress of habitat fragmentation in eastern Austria.     

Genetic diversity of the relict moss <Emphasis Type=Italic>Rhytidium rugosum</Emphasis> (Hypnales) in Europe inferred from the ITS region (nrDNA)

Post-glacial survival, potential migration routes, genetic diversity and phylogeography of the boreal moss species Rhytidium rugosum have been studied. This species is considered to be one of glacial relics of the wide but scattered Holarctic range. According to molecular data sampling from the selected European, American and Asian populations high genetic diversity of this species is present, even if this species is mostly sterile and produced sex organs extremely rarely and spread mostly asexually. Analysing the internal transcribed spacer (ITS) of the nuclear ribosomal DNA, it can be concluded that the populations of this species survived glaciations in various places in Europe and settled and re-settled present range space in various times from various refuges.     

Novel Algorithm for Non-Invasive Assessment of Fibrosis in NAFLD

Introduction

Various conditions of liver disease and the downsides of liver biopsy call for a non-invasive option to assess liver fibrosis. A non-invasive score would be especially useful to identify patients with slow advancing fibrotic processes, as in Non-Alcoholic Fatty Liver Disease (NAFLD), which should undergo histological examination for fibrosis.

Patients/Methods

Classic liver serum parameters, hyaluronic acid (HA) and cell death markers of 126 patients undergoing bariatric surgery for morbid obesity were analyzed by machine learning techniques (logistic regression, k-nearest neighbors, linear support vector machines, rule-based systems, decision trees and random forest (RF)). Specificity, sensitivity and accuracy of the evaluated datasets to predict fibrosis were assessed.

Results

None of the single parameters (ALT, AST, M30, M60, HA) did differ significantly between patients with a fibrosis score 1 or 2. However, combining these parameters using RFs reached 79% accuracy in fibrosis prediction with a sensitivity of more than 60% and specificity of 77%. Moreover, RFs identified the cell death markers M30 and M65 as more important for the decision than the classic liver parameters.

Conclusion

On the basis of serum parameters the generation of a fibrosis scoring system seems feasible, even when only marginally fibrotic tissue is available. Prospective evaluation of novel markers, i.e. cell death parameters, should be performed to identify an optimal set of fibrosis predictors.     

Heterotrophic soil respiration and soil carbon dynamics in the deciduous Hainich forest obtained by three approaches

Three different approaches were used to calculate heterotrophic soil respiration (Rh) and soil carbon dynamics in an old-growth deciduous forest in central Germany. A root and mycorrhiza exclosure experiment in the field separated auto- and heterotrophic soil respiration. It was compared to modeled heterotrophic respiration resulting from two different approaches: a modular component model of soil respiration calculated autotrophic and heterotrophic soil respiration with litter, climate and canopy photosynthesis as input variables. It was calibrated by independent soil respiration measurements in the field. A second model was calibrated by incubation of soil samples from different soil layers in the laboratory. In this case, the annual sum of Rh was calculated by an empirical model including response curves to temperature and a soil moisture. The three approaches showed good accordance during spring and summer and when the annual sums of Rh calculated by the two models were compared. Average Rh for the years 2002–2006 were 436 g C m^?2 year^?1 (field model) and 417 g C m^?2 year^?1 (lab-model), respectively. Differences between the approaches revealed specific limitations of each method. The average carbon balance of the Hainich forest soil was estimated to be between 1 and 35 g C m^?2 year^?1 depending on the model used and the averaging period. A comparison with nighttime data from eddy covariance (EC) showed that EC data were lower than modelled soil respiration in many situations. We conclude that better filter methods for EC nighttime data have to be developed.     

Intravenous tolerability of an acid vehicle in Sprague Dawley rats and beagle dogs after daily slow bolus injection and infusion for one hour during 14 days

To assess the tolerability of an acid vehicle to be used in toxicology studies, a low pH aqueous solution containing 16.4 mg/ml of citric acid, 4.2 mg/ml of disodium phosphate, 25 mg/ml of mannitol, adjusted with phosphoric acid/NaOH 1 M to pH 3 was daily administered intravenously to rats and dogs for 14 consecutive days. The dosing regimen consisted of a slow intravenous bolus injection given over 30 s (0.75 and 0.625 ml/kg, for rats and dogs, respectively) followed by intravenous infusion for one hour (3.75 and 2.75 ml/kg/h, for rats and dogs, respectively). In rats, the dose was administered via the lateral tail vein. In dogs, the intravenous bolus dose was administered via the vena cephalica, vena saphena or vena jugularis, whilst the infusion dose was given into the vena cephalica or vena saphena. In rats, administration of the vehicle was associated with clinical signs (occasional mild vocalization and agitation) which were considered to be due to local irritation during the dosing procedure. Nevertheless, only mild histopathological changes at the injection site were found, while no relevant clinical chemistry changes were found in this species. However, the vehicle caused significant vascular damage with thrombus formation in the dog. It is therefore concluded that this vehicle is suitable for 2-week rat toxicity studies, if carefully applied. The vehicle with its present regimen should not be used in dogs, in view of the prohibitive findings.     

Rhizobium nod genes are involved in the induction of two early nodulin genes in Vicia sativa root nodules

Nodulin gene expresison was studied in Vicia sativa (common vetch) root nodules induced by several Rhizobium and Agrobacterium strains. An Agrobacterium transconjugant containing a R. leguminosarum symplasmid instead of its Ti-plasmid, that was previously shown to form empty nodules on pea, induced nodules on Vicia roots in which nodule cells were infected with bacteria. In the Vicia nodules induced by this transconjugant, two so-called early nodulin genes were found to be expressed, whereas in the nodules formed on pea the expression of only one early nodulin gene was detected. In both cases the majority of the nodulin genes was not expressed.Apparently, an intracellular location of the bacteria is not sufficient for the induction of the majority of the nodulin genes. All nodulin genes were expressed in nodules induced by cured Rhizobium strains containing cosmid clones that have a 10 kb nod region of the sym-plasmid in common. Since in tumours no nodulin gene expression was found at all, the Agrobacterium chromosome does not contribute to the induction of nodulin genes. Therefore it is concluded that the signal for the induction of the expression of the two Vicia early nodulin genes is encoded by the nod-region, and the signal involved in the induction of all other nodulin genes has to be located outside the sym-plasmid, on the Rhizobium chromosome. The apparent difference in early nodulin gene expression between pea and Vicia is discussed in the light of the usefulness of Agrobacterium transconjugants in the study of nodulin gene expression.     

Neonatal steroids induce a down-regulation of tenascin-C and elastin and cause a deceleration of the first phase and an acceleration of the second phase of lung alveolarization

Pre- and postnatal corticosteroids are often used in perinatal medicine to improve pulmonary function in preterm infants. To mimic this clinical situation, newborn rats were treated systemically with dexamethasone (Dex), 0.1–0.01 mg/kg/day on days P1–P4. We hypothesized that postnatal Dex may have an impact on alveolarization by interfering with extracellular matrix proteins and cellular differentiation. Morphological alterations were observed on 3D images obtained by high-resolution synchrotron radiation X-ray tomographic microscopy. Alveolarization was quantified stereologically by estimating the formation of new septa between days P4 and P60. The parenchymal expression of tenascin-C (TNC), smooth muscle actin (SMA), and elastin was measured by immunofluorescence and gene expression for TNC by qRT-PCR. After Dex treatment, the first phase of alveolarization was significantly delayed between days P6 and P10, whereas the second phase was accelerated. Elastin and SMA expressions were delayed by Dex treatment, whereas TNC expression was delayed and prolonged. A short course of neonatal steroids impairs the first phase of alveolarization, most likely by altering the TNC and elastin expression. Due to an overshooting catch-up during the second phase of alveolarization, the differences disappear when the animals reach adulthood.     

Effect of salt and volume loading on the circulation in the leech,Hirudo medicinalis L.

Summary The effects of increased fluid volume in the closed vascular system on circulation were studied in the leech (Hirudo medicinalis) by intravascular pressure recordings and blood flow measurements.Significant increases in blood volume were achieved by crop loading with hyposmotic (72 mOsmol·kg^–1 H₂O) or hyperosmotic (300 mOsmol·kg^–1 H₂O) salt solutions or by infusion of isosmotic saline (200 mOsmol·kg^–1) into the vascular system.During the high-pressure (HIP) phase, which maintains the rear-to-front circulation, systolic blood pressure in the heart was not affected. An increase in systolic pressure in the heart was observed during the low-pressure (LOP) phase, which supplies the segmental circulation. Heart rate was not changed by crop loading with hyposmotic saline or by vascular infusion. Heart rate decreased after crop loading with hyperosmotic saline. Blood flow rate in the dorsal vessel was increased by crop loading with hyposmotic saline, but not after crop loading with hyperosmotic saline. In all cases the diameter of the dorsal vessel was not affected. A possible mechanism controlling blood pressure and blood flow in the vascular system is discussed.Abbreviations HIP-phase high-pressure phase - LOP-phase low-pressure phase - CNS central nervous system