Synthesis of O-β-D-Ribofuranosyl-(1″-2′)-adenosine-5″-O-phosphate

Abstract

The first synthesis of O-β-D-ribofuranosyl-(1″-2′)-adenosine-5″-O-phosphate starting from protected 2′-O-β-D-ribofuranosyladenosine has been performed.     

C11ORF24 Is a Novel Type I Membrane Protein That Cycles between the Golgi Apparatus and the Plasma Membrane in Rab6-Positive Vesicles

The Golgi apparatus is an intracellular compartment necessary for post-translational modification, sorting and transport of proteins. It plays a key role in mitotic entry through the Golgi mitotic checkpoint. In order to identify new proteins involved in the Golgi mitotic checkpoint, we combine the results of a knockdown screen for mitotic phenotypes and a localization screen. Using this approach, we identify a new Golgi protein C11ORF24 (NP_071733.1). We show that C11ORF24 has a signal peptide at the N-terminus and a transmembrane domain in the C-terminal region. C11ORF24 is localized on the Golgi apparatus and on the trans-Golgi network. A large part of the protein is present in the lumen of the Golgi apparatus whereas only a short tail extends into the cytosol. This cytosolic tail is well conserved in evolution. By FRAP experiments we show that the dynamics of C11ORF24 in the Golgi membrane are coherent with the presence of a transmembrane domain in the protein. C11ORF24 is not only present on the Golgi apparatus but also cycles to the plasma membrane via endosomes in a pH sensitive manner. Moreover, via video-microscopy studies we show that C11ORF24 is found on transport intermediates and is colocalized with the small GTPase RAB6, a GTPase involved in anterograde transport from the Golgi to the plasma membrane. Knocking down C11ORF24 does not lead to a mitotic phenotype or an intracellular transport defect in our hands. All together, these data suggest that C11ORF24 is present on the Golgi apparatus, transported to the plasma membrane and cycles back through the endosomes by way of RAB6 positive carriers.     

Occurrence of Fatty Acid Short‐Chain‐Alkyl Esters in Fruits of Celastraceae Plants

Small amounts of a mixture of fatty acid short‐chain‐alkyl esters (FASCAEs) were obtained from the fruits of twelve plant species of Celastraceae family, and in five of them the FASCAEs were present not only in the arils but also in the seeds. These mixtures contained 32 individual FASCAE species, which formed four separate fractions, viz. FA methyl, ethyl, isopropyl, and butyl esters (FAMEs, FAEEs, FAIPEs, and FABEs, resp.). The FASCAE acyl components included the residues of 16 individual C₁₄–C₂₄ saturated, mono‐, di‐, and trienoic FAs. Linoleic, oleic, and palmitic acids, and, in some cases, also α‐linolenic acid predominated in FAMEs and FAEEs, while myristic acid was predominant in FAIPEs. It can be suggested that, in the fruit arils of some plant species, FAMEs and FAEEs were formed at the expense of a same FA pool characteristic of a given species and were strongly different from FAIPEs and FABEs esters regarding the mechanism of their biosynthesis. However, as a whole, the qualitative and quantitative composition of various FASCAE fractions, as well as their FA composition, varied considerably depending on various factors. Therefore, separate FASCAE fractions seem to be synthesized from different FA pools other than those used for triacylglycerol formation.     

The Bacillus cereus Hbl and Nhe Tripartite Enterotoxin Components Assemble Sequentially on the Surface of Target Cells and Are Not Interchangeable

Bacillus cereus is a spore-forming, Gram-positive bacterium commonly associated with outbreaks of food poisoning. It is also known as an opportunistic pathogen causing clinical infections such as bacteremia, meningitis, pneumonia, and gas gangrene-like cutaneous infections, mostly in immunocompromised patients. B. cereus secretes a plethora of toxins of which four are associated with the symptoms of food poisoning. Two of these, the non-hemolytic enterotoxin Nhe and the hemolysin BL (Hbl) toxin, are predicted to be structurally similar and are unique in that they require the combined action of three toxin proteins to induce cell lysis. Despite their dominant role in disease, the molecular mechanism of their toxic function is still poorly understood. We report here that B. cereus strain ATCC 10876 harbors not only genes encoding Nhe, but also two copies of the hbl genes. We identified Hbl as the major secreted toxin responsible for inducing rapid cell lysis both in cultured cells and in an intraperitoneal mouse toxicity model. Antibody neutralization and deletion of Hbl-encoding genes resulted in significant reductions of cytotoxic activity. Microscopy studies with Chinese Hamster Ovary cells furthermore showed that pore formation by both Hbl and Nhe occurs through a stepwise, sequential binding of toxin components to the cell surface and to each other. This begins with binding of Hbl-B or NheC to the eukaryotic membrane, and is followed by the recruitment of Hbl-L₁ or NheB, respectively, followed by the corresponding third protein. Lastly, toxin component complementation studies indicate that although Hbl and Nhe can be expressed simultaneously and are predicted to be structurally similar, they are incompatible and cannot complement each other.     

Soil conservation practices for insect pest management in highly disturbed agroecosystems – a review

Decline in soil health is a serious worldwide problem that decreases complexity and stability of agricultural ecosystems, commonly making them more prone to outbreaks of herbivorous insect pests. Potato (Solanum tuberosum L., Solanaceae) and onion (Allium cepa L., Amaryllidaceae) production is currently characterized by high soil disturbance and heavy reliance on synthetic inputs, including insecticides. Evidence suggests that adopting soil conservation techniques often (but not always) increases mortality and decreases reproductive output for the major insect pests of these important vegetable crops. Known mechanisms responsible for such an effect include increases in density and activity of natural enemy populations, enhanced plant defenses, and modified physical characteristics of respective agricultural habitats. However, most research efforts focused on mulches and organic soil amendments, with additional research needed on elucidating effects and their mechanisms for conservation tillage, cover crops, and arbuscular mycorrhizae.     

A new weevil genus of the tribe Metrioxenini (Coleoptera: Belidae) in Eocene Baltic amber

A new genus, Succinometrioxena n. gen., and a new species, S. poinari n. sp. (Metrioxenini: Oxycoryninae), are described from Baltic amber. This new genus resembles the recent genus Wallacexena Legalov, 2009 but differs from it by the frons with horn-like tubercles on either side, the absence of a middle pronotal carina, teeth on the side of the pronotum, the anterior margin of the pronotum lacking teeth and two long carinae on the elytra. From the fossil genus Paltorhynchus Scudder, 1893 the present fossil differs by possessing a frons with horn-like tubercles on either side, the absence of a middle pronotal carina, teeth on the sides of the pronotum, two long carinae on the elytra and short temples. From the fossil genus Archimetrioxena Voss, 1953, the new genus differs by the frons with horn-like tubercles on either side, the absence of the two truncated elytral carinae, the sides of the pronotum with teeth, short temples and a larger body size.     

New species of the subfamily Cossoninae (Coleoptera: Curculionidae) in Dominican amber

The following new genus and seven new species of Cossonine weevils (Curculionidae: Cossoninae) are described from Dominican amber: Acamptus exilipes Poinar and Legalov, n. sp. (similar to A. rigidus), Caulophilus camptus Poinar and Legalov, n. sp. (similar to C. bennetti), Caulophilus elongatus Poinar and Legalov, n. sp. (similar to C. swensoni), Caulophilus ruidipunctus Poinar and Legalov, n. sp., Dryotribus pedanus Poinar and Legalov, n. sp. (similar to D. amplioculus), Ogygius obrieni Poinar and Legalov, n. gen., n. sp. (similar to the genus Apotrepus), and Stenotrupis pumilis Poinar and Legalov, n. sp. (similar to S. breviscapus). A key to the Dominican amber species of the genus Caulophilus is provided. Based on the extant weevil fauna, it appears that the Hispaniolan Cossonine were more diverse in the Tertiary than at present. http://zoobank.org/urn:lsid:zoobank.org:pub:14D1CFC0-4967-4D22-888B-0DE3E157D6BD     

New species of the genera Dryophthorus Germ. and Stenommatus Woll. (Coleoptera: Dryophthoridae) in Dominican amber

A new species of Dryophthorus and two new species of Stenommatus (Coleoptera: Dryophthoridae) are described from Dominican amber. Dryophthorus microtremus n. sp. is characterised by monochrome dorsal scales, nearly parallel sides of the pronotum, wide elytral intervals and the elytral apex only slightly narrowed. Stenommatus tanyrhinus n. sp. is characterised by an elongate rostrum 4.5 times as long as wide in the middle. S. leptorhinus is characterised by small body size, a thin rostrum 3.0 times as long as wide in the middle, wide striae and a narrow prosternal process. This brings the total number of species of Dryophthorini in Dominican amber to five, which is more than half the number of extant species.http://www.zoobank.org/urn:lsid:zoobank.org:pub:4329D0DA-57BD-4CEA-9464-ABD5D1577FC9     

Comparison of Complex DNA Mixtures with Generic Oligonucleotide Microchips

Abstract

The reproducibility of melting curves for repeated hybridizations of target DNA with generic oligonucleotide microchips is shown experimentally to depend on the character of matching between fragments of target DNA and immobilized oligonucleotides. The reproducibility of melting curves is higher for the perfect match duplexes and decreases as the number of mismatched pairs within duplexes increases. This effect was applied to the comparative analysis of complex DNA mixtures. We developed a scheme in which we can identify and discriminate between the probe oligonucleotides responsible for the distinctions between target DNA mixtures. A scheme is illustrated by comparing DNA mixtures corresponding to VD-J genes connected with populations of mRNAs CDR3 TCR Vb (T-cell receptor beta complementarity determining region 3) from the thymus and pancreas of NOD mice. Our results demonstrate that generic microchips can be applied efficiently to the analysis of DNA mixtures.     

Elastic properties of the growth-controlling outer cell walls of maize coleoptile epidermis

The effects of tensile stress and temperature on cell wall elasticity have been investigated in the outer cell walls of coleoptile epidermis of 4- and 6-day-old Zea mays L. seedlings. The change in tensile stress from 6 to 40 MPa caused the increase in cell wall elastic modulus from 0.4 to 3 GPa. Lowering the temperature from 30 to 4 °C resulted in instantaneous and reversible cell wall elongation of 0.3–0.5 ‰. At a given temperature and stress level, the wall elastic modulus of 6-day-old seedlings tended to be 30 % higher than that of 4-day-old plants. The relationship between cell wall elasticity and mechanical stress indicated that the stress distribution within the cell wall is highly uneven. The analysis of the effect of temperature on cell wall elastic strain showed that structural differences between crystalline and amorphous load-bearing polymers were not the only cause of the uneven stress distribution. Based on the results obtained by Hejnowicz and Borowska-Wykr?t (Planta 220:465–473, 2005), we suggested that the uneven stress distribution is partially related to the stress gradient between inner and outer layers of the cell wall.