Ultrastructure of grape leaf protoplasts in comparison with the source tissue

Based on their histochemical properties, protoplasts from grape leaf mesophyll were found to be viable and devoid of remnants of the cell wall. The results were confirmed by electron microscopy of sectioned and freeze-etched protoplast preparations. The only remarkable difference between cells in the intact tissue and the protoplasts was the occurrence of electron-dense globules in the cytoplasm of some individuals. These "osmiophilic bodies" are believed to be due to inevitable plasmolytic effects during wall removal and seem not to have any influence on protoplast CO2-assimilation, as has been demonstrated earlier. lt therefore appears that the vital structures remain intact and functional, in spite of the stress situation created by the isolation procedure.Ultrastruktur von Rebenblatt-Protoplasten und Ausgangsgewebe - ein VergleichAnhand histochemischer Nachweise, welche das Weiterfunktionieren der Plasmamembran als physiologische Barriere nach dem Isolationsprozeß bestätigen, können Mesophyllprotoplasten von Vitis vinifera L. als lebensfähig betrachtet werden. Dieser Befund, sowie das vollständige Fehlen von Zellwandresten, wurde durch elektronenmikroskopische Untersuchungen an Ultradünnschnitten bzw. Gefrierätzproben von Protoplastenpräparaten erhärtet. Das vereinzelte Auftreten von elektronendichten, runden Gebilden (sog. osmiophilic bodies) im Cytoplasma ist vermutlich eine der plasmolysierenden Wirkung des Isolationsmediums zuzuschreibende Erscheinung. Wie früher gezeigt, hat aber die verwendete Isolationsmethodik keinen negativen Einfluß auf den photosynthetischen C-Stoffwechsel der Protoplasten. Daraus geht hervor, daß die lebenswichtigen Zellkomponenten, trotz der isolationsbedingten Streßsituation, keinen irreversiblen und damit nachweisbaren, strukturellen oder funktionellen Beeinträchtigungen unterworfen sind

Evolutionary patchwork of an insecticidal toxin shared between plant-associated pseudomonads and the insect pathogens Photorhabdus and Xenorhabdus.

BACKGROUND: Root-colonizing fluorescent pseudomonads are known for their excellent abilities to protect plants against soil-borne fungal pathogens. Some of these bacteria produce an insecticidal toxin (Fit) suggesting that they may exploit insect hosts as a secondary niche. However, the ecological relevance of insect toxicity and the mechanisms driving the evolution of toxin production remain puzzling. RESULTS: Screening a large collection of plant-associated pseudomonads for insecticidal activity and presence of the Fit toxin revealed that Fit is highly indicative of insecticidal activity and predicts that Pseudomonas protegens and P. chlororaphis are exclusive Fit producers. A comparative evolutionary analysis of Fit toxin-producing Pseudomonas including the insect-pathogenic bacteria Photorhabdus and Xenorhadus, which produce the Fit related Mcf toxin, showed that fit genes are part of a dynamic genomic region with substantial presence/absence polymorphism and local variation in GC base composition. The patchy distribution and phylogenetic incongruence of fit genes indicate that the Fit cluster evolved via horizontal transfer, followed by functional integration of vertically transmitted genes, generating a unique Pseudomonas-specific insect toxin cluster. CONCLUSIONS: Our findings suggest that multiple independent evolutionary events led to formation of at least three versions of the Mcf/Fit toxin highlighting the dynamic nature of insect toxin evolution

Effects of 3^3He Impurity on Solid 4^4He Studied by Compound Torsional Oscillator

Frequency shifts and dissipations of a compound torsional oscillator induced by solid $^4$He samples containing $^3$He impurity concentrations ($x_3$ = 0.3, 3, 6, 12 and 25 in units of 10$^{-6}$) have been measured at two resonant mode frequencies ($f_1$ = 493 and $f_2$ = 1164 Hz) at temperatures ($T$) between 0.02 and 1.1 K. The fractional frequency shifts of the $f_1$ mode were much smaller than those of the $f_2$ mode. The observed frequency shifts continued to decrease as $T$ was increased above 0.3 K, and the conventional non-classical rotation inertia fraction was not well defined in all samples with $x_3 \geq$ 3 ppm. Temperatures where peaks in dissipation of the $f_2$ mode occurred were higher than those of the $f_1$ mode in all samples. The peak dissipation magnitudes of the $f_1$ mode was greater than those of the $f_2$ mode in all samples. The activation energy and the characteristic time ($\tau_0$) were extracted for each sample from an Arrhenius plot between mode frequencies and inverse peak temperatures. The average activation energy among all samples was 430 mK, and $\tau_0$ ranged from 2$\times 10^{-7}$ s to 5$\times 10^{-5}$ s in samples with $x_3$ = 0.3 to 25 ppm. The characteristic time increased in proportion to $x_3^{2/3}$. Observed temperature dependence of dissipation were consistent with those expected from a simple Debye relaxation model \emph{if} the dissipation peak magnitude was separately adjusted for each mode. Observed frequency shifts were greater than those expected from the model. The discrepancies between the observed and the model frequency shifts increased at the higher frequency mode.Comment: 10 pages, 10 figure

Engaging Religious Institutions to Address Racial Disparities in HIV/AIDS: A Case of Academic-Community Partnership

African Americans face the most severe burden of HIV among all racial and ethnic groups. Direct involvement of faith leaders and faith communities is increasingly suggested as a primary strategy to reduce HIV-related disparities, and Black churches are uniquely positioned to address HIV stigma, prevention, and care in African American communities. The authors describe an academic-community partnership to engage Black churches to address HIV in a predominantly African American, urban, southern Midwest location.  The opportunities, process, and challenges in forming this academic-community partnership with Black churches can be used to guide future efforts toward engaging faith institutions, academia, and other community partners in the fight against HIV.

Laser-Induced Fabrication of Metallic Interlayers and Patterns in Polyimide Films

Self-metallizing polyimide films are created by doping polyamic acid solutions with metallic ions and solubilizing agents. Upon creating a film, the film is exposed to coherent light for a specific time and then cured. The resulting film has been found to have a metallic surface layer and a metallic subsurface layer (interlayer). The layer separating the metallic layer has a uniform dispersion of small metal particulates within the polymer. The layer below the interlayer has larger metal particulates uniformly distributed within the polymer. By varying the intensity or time of exposure to the coherent light, three-dimensional control of metal formation within the film is provided

Metabolic effects of elevated temperature on organic acid degradation in ripening <em>Vitis vinifera</em> fruit

First published online: September 1, 2014Berries of the cultivated grapevine Vitis vinifera are notably responsive to temperature, which can influence fruit quality and hence the future compatibility of varieties with their current growing regions. Organic acids represent a key component of fruit organoleptic quality and their content is significantly influenced by temperature. The objectives of this study were to (i) manipulate thermal regimes to realistically capture warming-driven reduction of malate content in Shiraz berries, and (ii) investigate the mechanisms behind temperature-sensitive malate loss and the potential downstream effects on berry metabolism. In the field we compared untreated controls at ambient temperature with longer and milder warming (2-4 °C differential for three weeks; Experiment 1) or shorter and more severe warming (4-6 °C differential for 11 days; Experiment 2). We complemented field trials with control (25/15 °C) and elevated (35/20 °C) day/night temperature controlled-environment trials using potted vines (Experiment 3). Elevating maximum temperatures (4-10 °C above controls) during pre-véraison stages led to higher malate content, particularly with warmer nights. Heating at véraison and ripening stages reduced malate content, consistent with effects typically seen in warm vintages. However, when minimum temperatures were also raised by 4-6 °C, malate content was not reduced, suggesting that the regulation of malate metabolism differs during the day and night. Increased NAD-dependent malic enzyme activity and decreased phosphoenolpyruvate carboxylase and pyruvate kinase activities, as well as the accumulation of various amino acids and γ-aminobutyric acid, suggest enhanced anaplerotic capacity of the TCA cycle and a need for coping with decreased cytosolic pH in heated fruit.C. Sweetman, V. O. Sadras, R. D. Hancock, K. L. Soole and C. M. For

R2R - software to speed the depiction of aesthetic consensus RNA secondary structures

<p>Abstract</p> <p>Background</p> <p>With continuing identification of novel structured noncoding RNAs, there is an increasing need to create schematic diagrams showing the consensus features of these molecules. RNA structural diagrams are typically made either with general-purpose drawing programs like Adobe Illustrator, or with automated or interactive programs specific to RNA. Unfortunately, the use of applications like Illustrator is extremely time consuming, while existing RNA-specific programs produce figures that are useful, but usually not of the same aesthetic quality as those produced at great cost in Illustrator. Additionally, most existing RNA-specific applications are designed for drawing single RNA molecules, not consensus diagrams.</p> <p>Results</p> <p>We created R2R, a computer program that facilitates the generation of aesthetic and readable drawings of RNA consensus diagrams in a fraction of the time required with general-purpose drawing programs. Since the inference of a consensus RNA structure typically requires a multiple-sequence alignment, the R2R user annotates the alignment with commands directing the layout and annotation of the RNA. R2R creates SVG or PDF output that can be imported into Adobe Illustrator, Inkscape or CorelDRAW. R2R can be used to create consensus sequence and secondary structure models for novel RNA structures or to revise models when new representatives for known RNA classes become available. Although R2R does not currently have a graphical user interface, it has proven useful in our efforts to create 100 schematic models of distinct noncoding RNA classes.</p> <p>Conclusions</p> <p>R2R makes it possible to obtain high-quality drawings of the consensus sequence and structural models of many diverse RNA structures with a more practical amount of effort. R2R software is available at <url>http://breaker.research.yale.edu/R2R</url> and as an Additional file.</p