Structure-wise discrimination of adenine and guanine by proteins on the basis of their nonbonded interactions

We have analyzed the nonbonded interactions of the structurally similar moieties, adenine and guanine forming complexes with proteins. The results comprise (a) the amino acid–ligand atom preferences, (b) solvent accessibility of ligand atoms before and after complex formation with proteins, and (c) preferred amino acid residue atoms involved in the interactions. We have observed that the amino acid preferences involved in the hydrogen bonding interactions vary for adenine and guanine. The structural variation between the purine atoms is clearly reflected by their burial tendency in the solvent environment. Correlation of the mean amino acid preference values show the variation that exists between adenine and guanine preferences of all the amino acid residues. All our observations provide evidence for the discriminating nature of the proteins in recognizing adenine and guanine.     

Self-association of glutamic acid-rich fusion peptide analogs of influenza hemagglutinin in the membrane-mimic environments: Effects of positional difference of glutamic acids on side chain ionization constant and intra- and inter-peptide interactions deduced from NMR and gel electrophoresis measurements

Two glutamic acid-rich fusion peptide analogs of influenza hemagglutinin were synthesized to study the organization of the charged peptides in the membranous media. Fluorescence and gel electrophoresis experiments suggested a loose association between the monomers in the vesicles. A model was built which showed that a positional difference of 3, 7 and 4, 8 results in the exposure of Glu3 and Glu7 side chains to the apolar lipidic core. Supportive results include: first, pK_a values of two pH units higher than reference value in aqueous medium for Glu3 and Glu7 CγH, whereas the deviation of pK_a from the reference value for Glu4 and Glu8 CγH is substantially smaller; second, Hill coefficients of titration shift of these protons indicate anti-cooperativity for Glu3 and Glu7 side chain protons but less so for Glu4 and Glu8, implying a strong electrostatic interaction between Glu3 and Glu7 possibly resulting from their localization in an apolar environment; third, positive and larger titration shift for NH of Glu3 is observed compared to that of Glu4, suggesting stronger hydrogen bond between the NH and the carboxylic group of Glu3 than that of Glu4, consistent with higher degree of exposure to hydrophobic medium for the side chain of Glu3.     

Timing and magnitude of lumbar spine contribution to trunk forward bending and backward return in patients with acute low back pain

Alterations in the lumbo-pelvic coordination denote changes in neuromuscular control of trunk motion as well as load sharing between passive and active tissues in the lower back. Differences in timing and magnitude aspects of lumbo-pelvic coordination between patients with chronic low back pain (LBP) and asymptomatic individuals have been reported; yet, the literature on lumbo-pelvic coordination in patients with acute LBP is scant. A case-control study was conducted to explore the differences in timing and magnitude aspects of lumbo-pelvic coordination between females with (n=19) and without (n=19) acute LBP. Participants in each group completed one experimental session wherein they performed trunk forward bending and backward return at preferred and fast paces. The amount of lumbar contribution to trunk motion (as the magnitude aspect) as well as the mean absolute relative phase (MARP) and deviation phase (DP) between thoracic and pelvic rotations (as the timing aspect) of lumbo-pelvic coordination were calculated. The lumbar contribution to trunk motion in the 2nd and the 3rd quarters of both forward bending and backward return phases was significantly smaller in the patient than the control group. The MARP and the DP were smaller in the patient vs. the control group during entire motion. The reduced lumbar contribution to trunk motion as well as the more in-phase and less variable lumbo-pelvic coordination in patients with acute LBP compared to the asymptomatic controls is likely the result of a neuromuscular adaptation to reduce painful deformation and to protect injured lower back tissues.     

The role of histidine residues in the alpha toxin of Clostridium perfringens

The α -toxin (phospholipase C) of Clostridium perfringens has been reported to contain catalytically essential zinc ions We report here that histidine residues are essential for the co-ordination of these ion(s). Incubation of alpha toxin with diethylpyrocarbonate, a histidine modifying reagent, did not result in the loss of phospholipase C activity unless the protein was first incubated with EDTA, suggesting that zinc ions normally protect the susceptible histidine residues. When the amino acid sequences of three phospholipase C's were aligned, essential zinc binding histidine residues in the non-toxic B. cereus phospholipase C were found in similar positions in the toxic C. perfringens enzyme and the weakly toxic C. bifermentans phospholipase C.     

Factors affecting the responses of female Canada Geese to disturbance during incubation

During incubation, Canada Geese (Branta canadensis) and other waterfowl flush from their nests in response to the approach of potential predators and subsequently return to nests after the threat of predation has abated. Differences among individuals in their responses to such disturbance (e.g., how close a potential predator approaches before flushing) are thought to reflect variation in female age, the reproductive value of clutches, and nest site characteristics. From 26 April to 26 May 2010, we examined the possible effects of these factors on the behavior of female Canada Geese (B. c. interior) on Akimiski Island, Nunavut. We used humans as surrogate predators and flushing distance and return time after disturbance as response variables. The distance at which females flushed in response to approaching humans was best explained by the density of vegetation around nests, with females on better concealed nests flushing at closer distances. We also found a weak relationship between return time and clutch size, with females with larger clutches returning to nests sooner. The frequency of human visits did not affect either female flushing distance or return time. Female Canada Geese in our study appeared to minimize risk of their own injury or death (future reproductive potential) in escape decisions, whereas return times appeared to minimize the risk of nest predation (current reproductive investment).     

Chemically-Synthesized Gene Encoding Modified Human Superoxpde Dismutase: Its Construction,Expression and Properties of the Product

The gene encoding modified human superoxide dismutax (h-SOD) with 153 amino acid residues was constructed by chemical synthesis using the phosphoramidite method. The gene was designed so as to use bacterial codons for expression in prokaryotes and to introduce several unique restriction sites for further mutagenesis by the cassette exchange method. The distance between Shine-Dalgarno sequence and initiation codon was adjusted to maximum expression by using synthesized oligonucleotide. In addition, Cys 6 of h-SOD was changed to Ala to improve instability of native h-SOD.

Synthesized structural gene of h-SOD was expressed in E. coli after induction of isopropyl β-D-thiogalactoside by inserting the gene into the expression vector pKK223–3 having tac promoter. The gene that has 10 base pairs between Shine-Dalgarno sequence and initiation codøn showed the most efficient expression. The gene produced three active SOD isomers as revealed by chromatofocusing.

The main isomer was purified to homogeneity and characterized. The h-SOD-Ala⁶ showed similar properties to those of native h-SOD with respect to molecular weight, subunit structure, absorption spectrum. but the modified SOD was more resistant to heat denaturation than was native h-SOD; half-denaturing temperature was shifted by 10°C. Thus. the exchange of Cys 6 to Ala of h-SOD increased a stability of the enzyme.     

Peripheral-zone treatments for plum curculio management: validation in commercial apple orchards

A field trial was conducted using an oil formulation of the entomopathogenic fungus, Metarhizium flavoviride Gams & Rozyspal (Deuteromycotina: Hyphomycetes), to control the variegated grasshopper, Zonocerus variegatus L. (Orthoptera: Pyrgomorphidae). The influence of dispersal and secondary pick-up from the spray residue on mortality was assessed by sampling insects from the treated plots at intervals and monitoring disease levels during subsequent incubation in the laboratory. The cumulative mortality curves showed the pathogen to be highly effective, even in samples taken 8 days after spraying. A model was developed to link the influence of mortality, incubation, secondary pick-up of spores and insect dispersal, on the shape of these cumulative mortality curves. The model proved useful for data received from an experiment using small plot sizes, where insect dispersal is a big problem. The spray residue had an important influence on overall insect mortality. The model also indicated a difference in speed of kill between field and laboratory samples, suggesting an incubation delay in the field.     

Role of lysine residues of the Magnaporthe oryzae effector AvrPiz-t in effector- and PAMP-triggered immunity

Magnaporthe oryzae is an important fungal pathogen of both rice and wheat. However, how M. oryzae effectors modulate plant immunity is not fully understood. Previous studies have shown that the M. oryzae effector AvrPiz-t targets the host ubiquitin-proteasome system to manipulate plant defence. In return, two rice ubiquitin E3 ligases, APIP6 and APIP10, ubiquitinate AvrPiz-t for degradation. To determine how lysine residues contribute to the stability and function of AvrPiz-t, we generated double (K1,2R-AvrPiz-t), triple (K1,2,3R-AvrPiz-t) and lysine-free (LF-AvrPiz-t) mutants by mutating lysines into arginines in AvrPiz-t. LF-AvrPiz-t showed the highest protein accumulation when transiently expressed in rice protoplasts. When co-expressed with APIP10 in Nicotiana benthamiana, LF-AvrPiz-t was more stable than AvrPiz-t and was less able to degrade APIP10. The avirulence of LF-AvrPiz-t on Piz-t:HA plants was less than that of AvrPiz-t, which led to resistance reduction and lower accumulation of the Piz-t:HA protein after inoculation with the LF-AvrPiz-t-carrying isolate. Chitin- and flg22-induced production of reactive oxygen species (ROS) was higher in LF-AvrPiz-t than in AvrPiz-t transgenic plants. In addition, LF-AvrPiz-t transgenic plants were less susceptible than AvrPiz-t transgenic plants to a virulent isolate. Furthermore, both AvrPiz-t and LF-AvrPiz-t interacted with OsRac1, but the suppression of OsRac1-mediated ROS generation by LF-AvrPiz-t was significantly lower than that by AvrPiz-t. Together, these results suggest that the lysine residues of AvrPiz-t are required for its avirulence and virulence functions in rice.     

神农架常绿落叶阔叶混交林凋落物养分特征

凋落物是联结陆地生态系统植物与土壤养分的重要媒介,了解凋落物养分特征有助于理解陆地生态系统物质循环的机理。该研究于2015年收集了神农架地区常绿落叶阔叶混交林的新鲜凋落物及现存凋落物,测定其不同器官中大量元素(C、N、P、K、Ca、Mg)的含量,据此分析其养分含量、养分归还量、养分储量及化学计量比的特征。结果发现:该常绿落叶阔叶混交林新鲜凋落物的C、K养分含量显著高于现存凋落物,N、P、Ca、Mg养分含量显著低于现存凋落物;其凋落物大量元素的养分归还量及养分储量大小顺序均为C Ca N Mg K P,分别为1569.84、52.44、34.82、6.24、5.24、1.30 kg hm~(-2) a~(-1)及1835.29、87.87、51.17、12.12、3.90、1.95 kg hm~(-2) a~(-1);其新鲜凋落物及现存凋落物的C∶N∶P分别为1307.33∶27.73∶1及976.48∶26.77∶1,新鲜凋落物的C∶N、C∶P显著高于现存凋落物,N∶P无显著区别。研究表明,新鲜凋落物与现存凋落物养分含量之间的差异与不同元素在分解过程中的可淋溶性及生物固持等因素有关。该地区常绿落叶阔叶混交林凋落物养分归还量及养分储量相对于亚热带阔叶林平均水平较低;且显著低于喀斯特地区同类型森林,主要与其凋落物产量、降水量及植被类型有关。该森林生态系统新鲜凋落叶与中国及全球范围内阔叶树种凋落叶相比C∶N较低,C∶P、N∶P较高,这可能是由于该地区N沉降及P限制现象较为严重所致。     

Optimizing return‐on‐effort for coral nursery and outplanting practices to aid restoration of the Great Barrier Reef

Coral nursery and outplanting practices have grown in popularity worldwide for targeted restoration of degraded “high value” reef sites, and recovery of threatened taxa. Success of these practices is commonly gauged from coral propagule growth and survival, which fundamentally determines the return‐on‐effort (RRE) critical to the cost‐effectiveness and viability of restoration programs. In many cases, RRE has been optimized from past successes and failures, which therefore presents a major challenge for locations such as the Great Barrier Reef (GBR) where no local history of restoration exists to guide best practice. In establishing the first multi‐taxa coral nursery on the GBR (Opal Reef, February 2018), we constructed a novel scoring criterion from concurrent measurements of growth and survivorship to guide our relative RRE, including nursery propagule numbers (stock density). We initially retrieved RRE scores from a database of global restoration efforts to date (n = 246; 52 studies) to evaluate whether and how success commonly varied among coral taxa. We then retrieved RRE scores for Opal Reef using initial growth and survivorship data for six key coral taxa, to demonstrate that RRE scores were high for all taxa predominantly via high survivorship over winter. Repeated RRE scoring in summer is therefore needed to capture the full dynamic range of success where seasonal factors regulating growth versus survivorship differ. We discuss how RRE scoring can be easily adopted across restoration practices globally to standardize and benchmark success, but also as a tool to aid decision‐making in optimizing future propagation (and outplanting) efforts.