Molecular identification and phylogenetic position of Cuora yunnanensis

The Yunnan box turtle (Cuora yunnanensis, Boulenger, 1906), which has drawn much attention in conservation biology, was regarded as extinct since it was previously known only from 12 specimens collected in Yunnan, China, before 1908. Recently, live specimens have been discovered which are suggested to be C. yunnanensis. To determine whether the newly discovered specimens are really C. yunnanensis, we have established a molecular phylogeny, with a 1725-bp fragment of mitochondrial DNA, using samples from three live individuals of C. yunnanensis, together with sequence data from a museum specimen of C. yunnanensis (MNHN 1907.10) and other members of the genus Cuora. We found that the three newly discovered individuals and the old museum specimen of C. yunnanensis are very similar both in morphology and in mitochondrial DNA sequence, suggesting that the three new individuals are the very C. yunnanensis, and thus the species is not extinct. Our phylogenetic analysis also demonstrates that C. yunnanensis is not of recent hybrid origin, but rather represents a distinct evolutionary lineage.     

Asymmetric epoxidation of <Emphasis Type="Italic">α</Emphasis>,<Emphasis Type="Italic">β</Emphasis>-unsaturated ketones using <Emphasis Type="Italic">α</Emphasis>,<Emphasis Type="Italic">α</Emphasis>-diarylprolinols as catalysts

Asymmetric epoxidation of α,β-unsaturated ketones has been extensively studied and several important procedures have been developed in the last decade. This review addresses the most significant advances in asymmetric epoxidation of α,β-unsaturated ketones using proline-derived α,α-diarylprolinols as catalysts. Special attention has been paid to the enantioselective epoxidation of chalcones, α,β-unsaturated trifluoromethyl, trichloromethyl ketones and β,γ-unsaturated α-keto esters based on the reseach of our group.     

The R- and AR-indices： Complementing the h-index

Based on the foundation laid by the h-index we introduce and study the R- and AR-indices. These new indices eliminate some of the disadvantages of the h-index, especially when they are used in combina-tion with the h-index. The R-index measures the h-core’s citation intensity, while AR goes one step further and takes the age of publications into account. This allows for an index that can actually in-crease and decrease over time. We propose the pair (h, AR) as a meaningful indicator for research evaluation. We further prove a relation characterizing the h-index in the power law model.     

Transpositional behaviour of the Ds element in the Ac/Ds system in rice

Insertional mutagenesis based on maize Activator/Dissociator (Ac/Ds) transposons is becoming a ma- jor approach used to produce a saturated mutant collection in rice. In this research, Ds-T-DNA trans- formed homozygotes were crossed with Ac-T-DNA transformed homozygotes in order to establish an Ac/Ds transposon system in rice. The successive investigation of Ds transposition from F1 to F5 gen- erations indicated that the frequencies of germinal transposition increased over successive genera- tions and reached 54.2% in F3 generation. The Ds transposition pattern revealed that a Ds transposition induced an approximately 170-bp deletion of T-DNA sequence and another Ds transposition carried a 272-bp T-DNA sequence. Using thermal asymmetric interlaced PCR (TAIL-PCR), some flanking se- quences of the Ds element were amplified. Analyses of 17 Ds-flanking sequences showed that five Ds were inserted into gene regions. The Ds could transpose not only to the linked sites but also to the unlinked sites. The frequency of inter-chromosomal transposition of Ds was 33.3%.     

Osmotically stress-regulated the expression of glutathione peroxidase 3 in Arabidopsis

Gene expression of glutathione peroxidase 3 (ATGPX3) in response to osmotic stress was analyzed in Arabidopsis using ATGPX3 promoter-glucuronidase (GUS) transgenic plants. High levels of GUS ex- pression were detected under osmotic stress in ATGPX3 promoter-GUS transgenic plants. Compared with the wild type, the growth and development of ATGPX3 mutants (atgpx3-1) were more sensitive to mannitol. In addition, the expression of RD29A, ABI1, ABI2 and RbohD in atgpx3-1 was induced by ABA stress. These results suggest that ATGPX3 might be involved in the signal transduction of osmotic stress.     

The permeability effect of microcystin-RR on <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Bacillus subtilis</Emphasis>

Microcystins are a kind of cyclic hepatoxins produced by many species of cyanobacteria. Most previous work have been done on the toxic effects of microcystins on animals and plants. However, the reports about the effect of microcystins on microbial cells are very limited. In this work, the permeability of MC-RR on the cell outer membrane of Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis) was discussed. The permeability effect of MC-RR on the cell outer membrane of E. coli and B. subtilis under different concentrations was demonstrated by a rapid and sustained reduction in the A₆₇₅ values of lysozyme-treated cells. The decrease of the absorbance values showed a time- and dose-effect. The extravasations of protein and carbonhydrate increased with the increment of the treated-concentration of MC-RR. The results showed that MC-RR could increase the permeability of cell outer membranes of E. coli and B. subtilis. The synergistic effects of MC-RR and lysozyme on bacteria indicated that MC-RR might play an ecological role in bacteria in combination with other substances in some aquatic environments.     

Synthesis,characterization and photoluminescence of aluminum <Emphasis Type="Italic">N</Emphasis>-aryloxo functionalized <Emphasis Type="Italic">β</Emphasis>-ketoiminate complexes

The synthesis, characterization and luminescent properties of aluminum complexes containing a dianionic N-aryloxo functionalized β-ketoiminate ligand are presented. 4-(2-Hydroxy-5-R-phenyl)imino-2-pentanone (R = Me, L¹H₂; R = tert-butyl, L²H₂) ligands reacted with AlEt₃ in tetrahydrofuran to give the aluminum complexes (L¹AlEt)₂ (1) and (L²AlEt)₂ (2) in reasonable isolated yields. X-ray diffraction revealed that complexes 1 and 2 have solvent-free centrosymmetric dimeric structures, and each aluminum center has distorted trigonal bipyramidal geometry. At room temperature, complexes 1 and 2 exhibit blue photoluminescence in acetonitrile with maximum emission wavelengths of 419 and 413 nm, respectively.     

Molecular cloning, expression and biochemical property analysis of AtKP1, a kinesin gene from Arabidopsis thaliana

Kinesins are common in a variety of eukaryotic cells with diverse functions. A cDNA encoding a member of the Kinesin-14B subfamily is obtained using 3＇-RACE technology and named AtKP1 （for Arabidopsis kinesin protein 1）. This cDNA has a maximum open reading frame of 3.3 kb encoding a polypeptide of 1087 aa. Protein domain analysis shows that AtKP1 contains the motor domain and the calponin homology domain in the central and amino-terminal regions, respectively. The carboxyl-terminal region with 202 aa residues is diverse from other known kinesins. Northern blot analysis shows that AtKP1 is widely expressed at a higher level in seedlings than in mature plants. 2808 bp of the AtKP1 promoter region is cloned and fused to GUS. GUS expression driven by the AtKP1 promoter region shows that AtKP1 is mainly expressed in vasculature of young organs and young leaf trichomes, indicating that AtKP1 may participate in the differentiation or development of Arabidopsis thaliana vascular bundles and trichomes. A truncated AtKP1 protein containing the putative motor domain is expressed in E. coil and affinity-purified. In vitro characterizations indicate that the polypeptide has nucleotide-dependent microtubule-binding ability and microtubule-stimulated ATPase activity.     

Identification of <Emphasis Type="Italic">Fallopia</Emphasis> species based on the chloroplast <Emphasis Type="Italic">mat</Emphasis>K gene sequences

In order to develop an efficient identification method for Fallopia plants and drugs, molecular analysis of the partial matK gene sequences was performed on 6 Fallopia species. Based on the matK sequences, a phylogenetic tree was constructed, which showed that different populations of inter- and intra-species could be specified and distinguished. The matK gene sequences of the 6 Fallopia species were all found to be of 1 271 bp in length, with some nucleotide variations throughout the entire sequences. The nucleotide difference at position 1 041 could distinguish F. denticulata from others, while specific nucleotide at position 1 154 became identification markers for F. aubertii. Moreover, four specific marker sites for F. multiflora var. ciliinerve at positions 216, 224, 1 060 and 1 179, seven for F. convolvulus at 318, 765, 772, 874, 936, 952 and 1 036, and seven for F. dentate alata at 111, 192, 366, 450, 457, 1 032 and 1 074 were also observed. By detecting the marker nucleotides and analyzing the phylogenetic relationship, the botanical origins of five inspected drugs were determined, suggesting that matK sequences can be used for authenticating Fallopia plants and drugs.     

dr1127： A novel gene of Deinococcus radiodurans responsible for oxidative stress

The functional analysis of dr1127,a novel gene in Deinococcus radiodurans was performed in this pa-per. The dr1127 gene was found occasionally in our microarray and 2-DE gel experiments. Mutation of the dr1127 gene decreased the γ-radiation and H2O2 resistance of D. radiodurans,and weakened the scavenging abilities of cell extracts for free radicals (superoxide anion,hydrogen peroxide,and hy-droxyl radical). Further oxidative damage assays demonstrated that the purified DR1127 protein of D. radiodurans could bind to double stranded DNA in vitro and protect DNA from oxidative damage in this way. These results suggest that the dr1127 gene is an important gene that can maintain γ-radiation and oxidative resistance in D. radiodurans and may take part in the oxidative stress process.     

B-Valued Dyadic Derivative

The principles of the new maximal operator H＊ we defined are discussed. We prove that it is bounded from martingale Hardy-Lorentz L^Xp.q[0,1） to the Lorentz L^Xp.q[0,1） for 1/2〈 p〈∞, 0〈~ q ≤ ∞, where X is any Banach space. When the Banach space X has the RN property, the sequence dnHnf converges to f a.e. Meanwhile the convergence in L^Xp norm for 1≤p〈∞ is a consequence of that the family functions K （n∈N） is an approximate identity.     

Separation and Anti-Hantaan Virus Activity of Extracts from Alternanthera philoxcroides in vitro and in vivo

A water-soluble substance was extracted from the Chinese herb, Alternantheraphiloxcroides with hot water and alcohol. Aliquots of this initial extract were further fractionated by treatment with ether, ethyl acetate and alcohol respectively. The four extracts were assayed for anti-viral activity against three serum, Hantaan virus 114 （HV114）, HV435 and A9 strains. Results show that the four extracts are capable of inhibiting Hantaan virus propagation, of which extract No. 1 has the best efficiency. The three dosage of extract No. 1, which are used upon three Hantaan virus serum IC50, are 153, 157, 154 μg/mL. New-born mice were made to be infected with HV114 and then fed in vivo with extract No.l on the 3rd, 10th and 14th days after being infected by the virus. The treatment continued for 8 days with a dosage of 2.5 g/kg. Result shows that survival rates of mice were 75%, 50% and 0, respectively. The median time to death （MTDs） of the three groups were 37, 30, 23 days.     

A study on in vitro and in vivo bioactivity of nano hydroxyapatite/polymer biocomposite

When bioactive materials are implanted in vivo, a bone-like apatite layer can be found on their surfaces, which is critical to the establishment of bone-bonding between materials and living tissues. In this study, bone-like apatite formation in vitro and in vivo on surface of nano apatite/polyamide (n-HA/PA66) composite was investigated, and the interface between the implanted composite and surrounding bone tissue of rabbit were also examined. The results revealed that in both simulated body fluids (SBF) and dorsal muscles of rabbit, bone-like apatite could form on the biocomposite surface. When the samples were implanted in cortical bone, they combined directly with the natural bone without fibrous tissue in-between. The results showed that the n-HA/PA66 biocomposite had excellent bioactivity, which might be a good candidate for bone defect replacement.     

Accurate localization and excision of genomic islands in four strains of <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> and <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis>

Mobile genomic islands (GIs) can be excised from the chromosome, then form a circular intermediate and be reintegrated into the chromosome by the GI internal integrase. Some mobile GIs can also be transferred into a new receptor cell by transformation, conjugation, or transduction. The action sites of the integrase are usually flanked direct repeats (DRs) of the GIs. Accurate localization of the flanking sequences is a precondition for determining the mobility of the GI. Mobile GIs are generally associated with transfer RNAs (tRNAs). Based on the correlation between flanking sequences and tRNA sequences, the flanking sequences of 11 putative mobile GIs in Pseudomonas aeruginosa PAO1, P. aeruginosa PA14, P. fluorescens Pf-5 and P. fluorescens Pf0-1 were identified. Among the 11 GIs, Pf0-1GI-1 is responsible for benzoate degradation. PAO1GI-1, Pf5GI-2, Pf5GI-3, and Pf5GI-4 were confirmed experimentally to be excised from a chromosome to form a circular intermediate. The action sites of the integrases are these GIs direct repeats. Due to distinct DRs, cutting sites for the internal integrase of PAO1GI-1, Pf5GI-2, Pf5GI-3 and Pf5GI-4 were determined outside the T-loop of the tRNA^Gly gene, outside the anticodon loop of the tRNA^Ser gene and tRNA^Lys gene, and at the asymmetric 3′-end of the tRNA^Leu gene, respectively. PAO1GI-1 and other mobile GIs may be transferred into many different strains that belong to different phyla because of the clear flanking sequences. This study describes basic information about the action sites of the integrases, assesses the mobility of GIs, and can help design and transfer mobile GIs to candidate strains.     

Asymmetric intramolecular oxa-Michael addition of activated <Emphasis Type="Italic">α,β</Emphasis>-unsaturated ketones by chiral <Emphasis Type="Italic">N</Emphasis>-triflyl phosphoramide

Asymmetric intramolecular oxa-Michael addition of activated α,β-unsaturated ketones by chiral N-triflyl phosphoramide was realized. The flavanone products can be synthesized with excellent yields (50%–95%) and up to 74% ee.     

Genetic and physical mapping of AvrPi7, a novel avirulence gene of Magnaporthe oryzae using physical position-ready markers

Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is one of the most devastating crop diseases worldwide. The avirulence gene corresponding to rice blast resistance gene Pi7 in field isolate CHL346 was inherited as a single gene, designated AvrPi7, in a segregating population consisting of 189 ascospore progenies derived from a cross between field isolates CHL346 and CHL42. In order to determine the chromosomal location of the AvrPi7 locus, a total of 121 simple sequence repeat (SSR) markers were developed based on the whole-genome sequence of reference isolate 70-15 of M. oryzae. Linkage analysis of the locus with these SSR markers showed that eight SSR markers on chromosome 1 were linked to the locus, among which the closest flanking markers MS1-9 and MS1-15 were 3.2 and 16.4 cM from the locus, respectively. For fine mapping, additional PCR-based makers including eight SSR markers and three candidate avirulence gene (CAG) markers were developed in the region flanking both markers. The AvrPi7 locus was genetically delimited within a 1.6-cM region flanked by markers MS1-21 and MS1-22, and co-segregated with the marker CAG2. To construct a physical map of the AvrPi7 locus, molecular markers linked to the Avr gene were mapped on the supercontigs of the ref-erence isolate 70-15 through bioinformation analysis (BIA). Consequently, the AvrPi7 locus was delim-ited to a 75-kb interval flanked by markers MS1-21 and MS1-22 based on the reference sequence. Merodiploids observed in this study are also discussed.