Repeated freeze–thaw cycles induce embolism in drought stressed conifers (Norway spruce,stone pine)

Freezing and thawing lead to xylem embolism when gas bubbles caused by ice formation expand during the thaw process. However, previous experimental studies indicated that conifers are resistant to freezing-induced embolism, unless xylem pressure becomes very negative during the freezing. In this study, we show that conifers experienced freezing-induced embolism when exposed to repeated freeze-thaw cycles and simultaneously to drought. Simulating conditions at the alpine timberline (128 days with freeze-thaw events and thawing rates of up to 9.5 K h(-1) in the xylem of exposed twigs during winter), young trees of Norway spruce [Picea abies (L.) Karst.] and stone pine (Pinus cembra L.) were exposed to 50 and 100 freeze-thaw cycles. This treatment caused a significant increase in embolism rates in drought-stressed samples. Upon 100 freeze-thaw cycles, vulnerability thresholds (50% loss of conductivity) were shifted 1.8 MPa (Norway spruce) and 0.8 MPa (stone pine) towards less negative water potentials. The results demonstrate that freeze-thaw cycles are a possible reason for winter-embolism in conifers observed in several field studies. Freezing-induced embolism may contribute to the altitudinal limits of conifers.     

Molecular mapping of QTLs for Fusarium head blight resistance in spring wheat. II. Resistance to fungal penetration and spread

Fusarium head blight (FHB, scab) causes severe yield and quality losses, but the most serious concern is the mycotoxin contamination of cereal food and feed. The cultivation of resistant varieties may contribute to integrated control of this fungal disease. Breeding for FHB resistance by conventional selection is feasible, but tedious and expensive. The aim of this work was to detect QTLs for combined type I and type II resistance against FHB and estimate their effects in comparison to the QTLs identified previously for type II resistance. A population of 364, F1 derived doubled-haploid (DH) lines from the cross 'CM-82036' (resistant)/'Remus' (susceptible) was evaluated for components of FHB resistance during 2 years under field conditions. Plants were inoculated at anthesis with a conidial suspension of Fusarium graminearum or Fusarium culmorum. The crop was kept wet for 20 h after inoculation by mist-irrigation. Disease severity was assessed by visual scoring. Initial QTL analysis was performed on 239 randomly chosen DH lines and extended to 361 lines for putative QTL regions. Different marker types were applied, with an emphasis on PCR markers. Analysis of variance, as well as simple and composite interval mapping, revealed that two genomic regions were significantly associated with FHB resistance. The two QTLs on chromosomes 3B (Qfhs.ndsu-3BS) and 5A (Qfhs.ifa-5A) explained 29 and 20% of the phenotypic variance, respectively, for visual FHB severity. Qfhs.ndsu-3BS appeared to be associated mainly with resistance to fungal spread, and Qfhs.ifa-5A primarily with resistance to fungal penetration. Both QTL regions were tagged with flanking SSR markers. These results indicate that FHB resistance was under the control of two major QTLs operating together with unknown numbers of minor genes. Marker-assisted selection for these two major QTLs appears feasible and should accelerate the development of resistant and locally adapted wheat cultivars.     

Average hydroxyapatite concentration is uniform in the extracollagenous ultrastructure of mineralized tissues: evidence at the 1–10-μm scale

At the ultrastructural observation scale of fully mineralized tissues (l=1-10 mum), transmission electron micrographs (TEM) reveal that hydroxyapatite (HA) is situated both within the fibrils and extrafibrillarly, and that the majority of HA lies outside the fibrils. The extrafibrillar amount of HA varies from tissue to tissue. By means of mathematical modeling, we here provide strong indications that there exists a physical quantity that is the same inside and outside the fibrils, for all different fully mineralized tissues. This quantity is the average mineral concentration in the non-collagenous space. This space is the sum of the extrafibrillar volume and of the volume of the fibrils that is not occupied by collagen molecules. Two independent sets of experimental observations covering a large range of tissue mass densities establish the relevance of our proposition: (i) mass density measurements and diffraction spacing measurements, re-analyzed through a dimensionally consistent packing model; (ii) optical density measurements of TEMs. The aforementioned average uniform HA-concentration in the extracollagenous space of the ultrastructure may emphasize the putative role played by a number of non-collagenous organic molecules in providing the chemical boundary conditions for mineralization of HA in the extracollagenous space. The probable existence of an average uniform extracollagenous HA concentration has far-reaching consequences for the mechanical behavior of mineralized tissues.     

<Emphasis Type="Italic">Aspergillus niger</Emphasis> citric acid accumulation: do we understand this well working black box?

This Mini-Review summarizes the current knowledge on the biochemical and physiological events leading to massive citric acid accumulation by Aspergillus niger under industrially comparable conditions, thereby particularly emphasizing the roles of glycolytic flux and its control, excretion of citric acid from the mitochondria and the cytosol, and the critical fermentation variables. The potential of novel techniques for metabolic analysis and genomic approaches in understanding this fermentation is also discussed.     

Contents of a storage pit from late Bronze Age Stillfried,Austria: another record of the "new" glume wheat

Since Jones et al. (2000) drew attention to a "new" type of glume wheat from Neolithic and Bronze Age sites in northern Greece, several finds of this morphologically distinct tetraploid wheat form have been made across central and southeastern Europe. Charred remains of this wheat, dating from 819–1031 cal b.c., have also been discovered in a storage pit at late Bronze Age Stillfried, eastern Austria. As both chaff and grains were found, it was not only possible to match the diagnostic features of the spikelet bases to the "new" form, but also to examine the grains, which are strikingly long, slender and flat. A dorsal ridge is absent and there is no hump above the embryo. The embryo angle is relatively low and compression lines are much more distinct. Within the Stillfried store "new" glume wheat grains were also easily separable from two-grained einkorn and spelt grains. The morphology of the grains is not inconsistent with the suggestion that the "new" type glume wheat might correspond to modern Triticum timopheevi. In Stillfried "new" glume wheat was grown as a winter crop, and it seems to have been cultivated as a maslin (mixed crop) together with T. monococcum (einkorn).     

Quantitative measurement of cell migration using time-lapse videomicroscopy and non-linear system analysis

Epithelial cells of the mammary gland possess the inherent capacity to form epithelial monolayers in vitro. This requires coordination of cell migration, cell-cell contact formation, and cell proliferation. Using time-lapse phase contrast videomicroscopy we have observed mammary gland epithelial cells over different time scales. We show the generation of a complete polarized epithelial monolayer in real-time, starting from a few cells. We subsequently concentrated on the early stages of this process by tracking epithelial cells during phases of polarized migration. We performed migration analysis using fractal measures. With this technology the structure of seemingly random processes not accessible to the usual methods of linear analysis can be measured. As a control and proof of principle approach we applied infection of cells with an adenoviral vector, which is used as a gene targeting vector for many applications. Infection markedly influenced the patterns of migratory behavior. We, therefore, believe that time-lapse videomicroscopy in combination with fractal analysis can contribute to differential characterization of distinct cellular migration patterns. This will be useful in situations of long-term alterations in cell culture systems.     

Role of B7-CD28/CTLA-4 costimulation and NF-kappa B in allergen-induced T cell chemotaxis by IL-16 and RANTES

The mechanisms that cause T cell recruitment into inflamed airways of asthmatic individuals are poorly understood. It has been shown previously that both natural exposure to allergen and challenge in the laboratory induce T cell accumulation in the bronchial mucosa of sensitized asthmatics. To study the mechanisms involved in this process, we have used an explant model in which bronchial biopsies taken from mild atopic asthmatic volunteers during fiberoptic bronchoscopy were stimulated in culture for 24 h by the common aeroallergen house dust mite (Dermatophagoides pteronyssinus (Der p)). Analysis of culture supernatants showed that stimulation with Der p significantly enhanced both the generation of T cell chemotactic activity by the mucosal tissue, as assayed in microchemotaxis chambers, and the production of IL-16 and RANTES. Neutralization experiments showed that IL-16 contributed more to the chemotactic activity than RANTES. The fusion protein CTLA-4-Ig, blocking B7:CD28 costimulation, and dexamethasone both significantly reduced the ex vivo production of chemotactic activity and release of IL-16 and RANTES. The proteasome inhibitor Cbz-Ile-Glu(OtBu)-Ala-leucinal also had a significant inhibitory effect on T cell chemotactic activity and IL-16 but not RANTES generation, indicating a role for nuclear factor NF kappa B activation. These results indicate that allergen stimulates cells within the bronchial mucosa to increase IL-16 and RANTES release, both of which contribute to T cell accumulation in asthmatic airways. The allergen-induced chemotactic activity is dependent on cell activation via CD28 and involves, at least partly, NF-kappa B.     

Ciboulot regulates actin assembly during Drosophila brain metamorphosis

A dynamic actin cytoskeleton is essential for the remodeling of cell shape during development, but the specific roles of many actin partners remain unclear. Here we characterize a novel actin binding protein, Ciboulot (Cib), which plays a major role in axonal growth during Drosophila brain metamorphosis. Loss of Cib function leads to axonal growth defects in the central brain, while overexpression of the gene during development leads to overgrown projections. The Cib protein displays strong sequence similarity to beta-thymosins but has biochemical properties like profilin: the Cib-actin complex participates in actin filament assembly exclusively at the barbed end, and Cib enhances actin-based motility in vitro. Genetic experiments show that Cib and the Drosophila profilin protein Chickadee (Chic) cooperate in central brain metamorphosis.     

Src Family Tyrosine Kinase Regulates Intracellular pH in Cardiomyocytes

The Anion Cl⁻/HCO₃⁻ Exchangers AE1, AE2, and AE3 are membrane pH regulatory ion transporters ubiquitously expressed in vertebrate tissues. Besides relieving intracellular alkaline and CO₂ loads, the AEs have an important function during development and cell death and play a central role in such cellular properties as cell shape, metabolism, and contractility. The activity of AE(s) are regulated by neurohormones. However, little is known as to the intracellular signal transduction pathways that underlie this modulation. We show here that, in cardiomyocytes that express both AE1 and AE3, the purinergic agonist, ATP, triggers activation of anion exchange. The AE activation is observed in cells in which AE3 expression was blocked but not in cells microinjected with neutralizing anti-AE1 antibodies. ATP induces tyrosine phosphorylation of AE1, activation of the tyrosine kinase Fyn, and association of both Fyn and FAK with AE1. Inhibition of Src family kinases in vivo by genistein, herbimycin A, or ST638 prevents purinergic activation of AE1. Microinjection of either anti-Cst.1 antibody or recombinant CSK, both of which prevent activation of Src family kinase, significantly decreases ATP-induced activation of AE. Microinjection of an anti-FAK antibody as well as expression in cardiomyocytes of Phe397 FAK dominant negative mutant, also prevents purinergic activation of AE. Therefore, tyrosine kinases play a key role in acute regulation of intracellular pH and thus in cell function including excitation–contraction coupling of the myocardium.