Crystal structure determination of a neutral neurotoxin BmK M4 from Buthus martensii Karsch at 0.20 nm

BmK M4 is a neutral neurotoxin in the BmK toxin series.It is medially toxic and belongs to group III α-toxins.The purified sample was crystallized in rhombic space group P61.Using an X-ray diffraction technique,the crystal structure of BmK M4 was revealed by molecular replacement at 0.20 nm resolution.The model was refined.The final crystallographic R factor was 0.142 and the free R factor was 0.173.The root mean square deviation is 0.001 5 nm for the bond length and 1.753°for the bond angles.64 water molecules were added to the asymmetric unit.The refined structure showed an unusual non-prolyl cis peptide bond at residue 10.The structure was compared with group II α-toxin BmK M8 (an acidic,weak toxin).The potential structural implications of the cis peptide bond were discussed.     

The phylogenetic position of the zokors (Myospalacinae) and comments on the families of muroids (Rodentia)

Recent molecular studies have concluded that the genus Myospalax evolved from within the rodent subfamily Cricetinae. This conclusion was tested using the complete sequences from the mitochondrial 12S rRNA and cytochrome b genes. Based on our analyses, Myospalax appears to be sister to a clade containing the subfamilies Spalacinae and Rhizomyinae, and all three of these lineages appear to be basal to the superfamily Muroidea. Based on the position of these three lineages, we suggest that they be placed in a distinct family, the Spalacidae, rather than subsumed as subfamilies in the family Muridae. Finally, our analyses suggest that the earlier placement of Myospalax as a member of the Cricetinae is the result of a single misidentified specimen, which was not a Myospalax.     

In vitro HIV-1 selective integration into the target sequence and decoy-effect of the modified sequence

Although there have been a few reports that the HIV-1 genome can be selectively integrated into the genomic DNA of cultured host cell, the biochemistry of integration selectivity has not been fully understood. We modified the in vitro integration reaction protocol and developed a reaction system with higher efficiency. We used a substrate repeat, 5'-(GTCCCTTCCCAGT)(n)(ACTGGGAAGGGAC)(n)-3', and a modified sequence DNA ligated into a circular plasmid. CAGT and ACTG (shown in italics in the above sequence) in the repeat units originated from the HIV-1 proviral genome ends. Following the incubation of the HIV-1 genome end cDNA and recombinant integrase for the formation of the pre-integration (PI) complex, substrate DNA was reacted with this complex. It was confirmed that the integration selectively occurred in the middle segment of the repeat sequence. In addition, integration frequency and selectivity were positively correlated with repeat number n. On the other hand, both frequency and selectivity decreased markedly when using sequences with deletion of CAGT in the middle position of the original target sequence. Moreover, on incubation with the deleted DNAs and original sequence, the integration efficiency and selectivity for the original target sequence were significantly reduced, which indicated interference effects by the deleted sequence DNAs. Efficiency and selectivity were also found to vary discontinuously with changes in manganese dichloride concentration in the reaction buffer, probably due to its influence on the secondary structure of substrate DNA. Finally, integrase was found to form oligomers on the binding site and substrate DNA formed a loop-like structure. In conclusion, there is a considerable selectivity in HIV-integration into the specified sequence; however, similar DNA sequences can interfere with the integration process, and it is therefore difficult for in vivo integration to occur selectively in the actual host genome DNA.     

Site fidelity and association patterns of Indo-Pacific humpback dolphins off the east coast of Zhanjiang,China

This study examined the population of Indo-Pacific humpback dolphins, Sousa chinensis, inhabiting the waters off the east coast of Zhanjiang, China. A total of 116 dolphins were identified during 147 boat-based surveys, completed between June 2005 and June 2007. Abundance estimates indicated that a small population of 268 dolphins (95% CI = 189–413) inhabited this coastal area. The sighting frequencies of identified dolphins varied between one and five when the 2006 and 2007 data were organized into six occasions. Twenty-three percent (27 individuals) of the cataloged dolphins were identified in more than one calendar year. The two animals (ZJ001, ZJ011) with the highest degree of site fidelity were present in the study area for a period of 23 months. The most frequently (15 times) sighted dolphin (ZJ046) was recorded in nine (56%) of the 16 months surveyed. This evidence indicates the interannual site fidelity of S. chinensis to the area off the eastern coast of Zhanjiang City. Range sizes of eight identified individuals calculated by the minimum convex polygon varied from 2.07 to 331.20 km². Associations between 34 individuals sighted at least three times and more than three times were measured by the half-weight index and SOCPROG program. The majority of these dolphins (80.57%) were not seen together during surveys and the mean association rate between dyads was low at 0.05. Temporal analyses for all the individuals were conducted using a lagged association rate. The results indicated a non-random social structure made by constant companions.     

Timing of mutation in hemagglutinins from influenza A virus by means of unpredictable portion of amino-acid pair and fast Fourier transform

In this study, we calculate the unpredictable portion of amino-acid pairs, which has been developed by us over the last several years, of 1201 hemagglutinins from influenza A viruses dated from 1918 to 2004 in order to compare them with respect to subtypes, species, and years. After noticing the fluctuations of unpredictable portion along the time course, we use the fast Fourier transform to find the mutation periodicity of hemagglutinins. Then we estimate our position at the current cycle of hemagglutinin evolutionary process to determine how many years remain before the next outbreak of influenza and bird flu. Finally, we use the trend line and channel to outlook the hemagglutinins for the next half a century. As our study covers almost all the full-length amino-acid sequences of hemagglutinins from various influenza A viruses, the conclusion will be valid for years until the number of hemagglutinins in protein databank will be significantly increased.     

Curcumin induces apoptosis through mitochondrial hyperpolarization and mtDNA damage in human hepatoma G2 cells

Curcumin, a major pigment of turmeric, is a natural antioxidant possessing a variety of pharmacological activities and therapeutic properties. But its mechanisms are unknown. In our previous study, we found that a 2-h exposure to curcumin induced DNA damage to both the mitochondrial DNA (mtDNA) and the nuclear DNA (nDNA) in HepG2 cells and that mtDNA damage was more extensive than nDNA damage. Therefore, experiments were initiated to evaluate the role of mtDNA damage in curcumin-induced apoptosis. The results demonstrated that HepG2 cells challenged with curcumin for 1 h showed a transient elevation of the mitochondrial membrane potential (DeltaPsim), followed by cytochrome c release into the cytosol and disruption of DeltaPsim after 6 h exposure to curcumin. Apoptosis was detected by Hoechst 33342 and annexin V/PI assay after 10 h treatment. Interestingly, the expression of Bcl-2 remained unchanged. A resistance to apoptosis for the corresponding rho0 counterparts confirmed a critical dependency for mitochondria during the induction of apoptosis in HepG2 cells mediated by curcumin. The effects of PEG-SOD in protecting against curcumin-induced cytotoxicity suggest that curcumin-induced cytotoxicity is directly dependent on superoxide anion O2- production. These data suggest that mitochondrial hyperpolarization is a prerequisite for curcumin-induced apoptosis and that mtDNA damage is the initial event triggering a chain of events leading to apoptosis in HepG2 cells.     

植物光诱导延迟荧光测量的影响因素研究

光合作用是地球上最重要的化学反应,植物内源性光诱导延迟荧光是光合作用原初过程中光系统Ⅱ作用中心P680处电荷分离效率的内在探针。延迟荧光除了受植物本身及其生长发育状况有关外,还受到其他很多环境及测量方面的影响,所以为了更好地利用延迟荧光特性技术研究植物生理特性,就必须对测量参数指标做合理的优化。本文从影响延迟荧光的激发光源的光强,激发时间及外界环境温度出发,研究延迟荧光的变化特性,为延迟荧光在植物生理特性方面的研究提供合理的理论依据。