[Experimental] List of manuscripts available for review volunteers
8 manuscripts available for review volunteers
February 18, 2017
Aerobic anoxygenic phototrophic bacteria (AAPB) containing bacteriochlorophyll a (BChl a) are photoheterotrophic prokaryotes that are a widely distributed functional bacterial group due to their obligately aerobic and facultative photoheterotrophic abilities. Here, we made a comparison of AAPB community structures in different marine functional zones in the surface water of the Zhoushan Archipelago Sea Area using high-throughput sequencing based on the pufM gene that encodes the M subunit of the light reaction centre complex. A total of 268,214 clean reads were obtained from the sampling stations. The sequences were divided into 5876 OTUs (97% cut-off value). Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria phyla were noted in this study. Proteobacteria phylum accounted for the major proportion and existed at all sites. Roseobacter-like AAPB was the most abundant genus, in Dongji Island (DJ) and Gouqi Island (GQ). Our results demonstrated that the structure of the AAPB community had different distribution patterns within the Zhoushan Archipelago Sea Area. Furthermore, we found that the diversity of AAPB was controlled by complex environmental factors, which might explain the difficulties encountered when predicting the distribution of total AAPB in aquatic ecosystems.
February 17, 2017

Objective: The purpose of this study was to demonstrate the effects of the achievement motive on important variables such as the purpose in life and social participation with objective physical functions in predicting within-person fluctuations and between-person differences using longitudinal research in community-dwelling Japanese elderly people.

Methods: The final dataset consisted of 227 persons (men: 109, women: 118) from day-service centers through testing at three time points. The tests comprised a questionnaire on the achievement motive, purpose in life and so on and the measurement of physical functions. We verified the following hypothesis model; 1) achievement motive works on improvement of the purpose in life, social participation, self-efficacy, and role expectation, 2) social participation and role expectation improve the purpose in life, 3) hobbies and going out affect the purpose in life, 4) social participation and physical function affect the increase in going out, 5) achievement motive and social participation improve physical function and increase role expectation and hobbies, and 6) self-efficacy improves social participation and the purpose in life. Our hypothesized model was based on previous research and was analyzed using a multilevel structural equation modeling approach.

Results: The modified hypothesis model without hobby, grip strength, and gait speed exhibited an adequate model fit: comparative fit index = 0.904, Tucker-Lewis index = 0.775, root mean square error of approximation = 0.064, Akaike’s information criterion = 25201.17, Bayesian information criterion = 25389.59, and adjusted BIC = 25256.24. Within level, the achievement motive had significant direct effects on the purpose in life, social participation, and self-efficacy. In addition, there were significant indirect effects of the achievement motive on the purpose in life through social participation and on social participation through self-efficacy. Between level, the achievement motive had significant direct effects on all variables and significant indirect effects on the purpose in life, social participation, and going out.

Conclusion: Our findings highlight the implications of achievement motive for the purpose in life, social participation and self-efficacy of the participation at the within person and between people levels. Consequently, we understood that the achievement motive could strongly affect between-person differences more than within-person fluctuations during a short period of six months.

February 12, 2017
WRKY proteins, comprising one of the largest transcription factor families in plant kingdom, play crucial roles in the plant development and stress responses. Despite several studies on WRKYs in wheat (Triticum aestivum L.) were investigated, functional annotation information about wheat WRKYs was limited. Here, 171 TaWRKY transcription factors (TFs) were identified from the whole wheat genome and compared with proteins from other 19 species representing nine major plant lineages. A phylogenetic analysis, coupled with gene structure analysis and motif determination, divided these TaWRKYs into seven subgroups (Group I, IIa-e, III). Chromosomal location showed that the most TaWRKY genes were enriched on four chromosomes, especially on chromosome 3B, and 85 (49.7%) genes were either tandem (5) or segmental duplication (80), which suggested that though tandem duplication has contributed to the expansion of TaWRKY family, segmental duplication probably played a more pivotal role. The cis-acting elements analysis revealed putative functions of WRKYs in wheat during development as well as under numerous biotic and abiotic stresses. Finally, the expression of TaWRKY genes in flag leaves, glumes and lemmas under water stress were analyzed, and we found different TaWRKY genes preferentially express in specific tissue during the grain-filling stage. Our results provide a more extensive knowledge on TaWRKYs, which helps to complete the information of WRKY gene family in wheat, and also contribute to screen more candidate genes for further investigation on function characterization of WRKYs under various stresses.
February 6, 2017

Objective: The trabecular meshwork (TM) is the primary substrate of outflow resistance in glaucomatous eyes. Repopulating diseased TM with fresh, functional TM cells might represent a novel therapeutic breakthrough. Various decellularized TM scaffolds were developed by ablating existing cells with suicide gene therapy or saponin, but always with incomplete cell removal or dissolve the extracellular matrix. We hypothesized that a chemical-free, freeze-thaw method would be able to produce a fully decellularized TM scaffold for cell transplantation.

Materials and Methods: We obtained 24 porcine eyes from a local abattoir, dissected and mounted them in an anterior segment perfusion and pressure transduction system within two hours of sacrifice. After they stabilized for 72 hours, eight eyes each were assigned to freeze-thaw (F) ablation (-80°C×2), to 0.02% saponin (S) treatment, or the control group (C), respectively. The trabecular meshwork was transduced with an eGFP expressing feline immunodeficiency viral (FIV) vector and tracked via fluorescent microscopy to confirm ablation. Following treatment, the eyes were perfused with standard tissue culture medium for 180 hours. We assessed histological changes by hematoxylin and eosin staining. TM cell viability was evaluated with a calcein AM/propidium iodide (PI) assay. We measured IOP and modeled it with a linear mixed effects model using a B-spline function of time with 5 degrees of freedom.

Results: F and S experienced a similar IOP reduction by 30% from baseline (P=0.64). IOP reduction of about 30% occurred in F within 24 hours and in S within 48 hours. Live visualization of eGFP demonstrated that F conferred a complete ablation of all TM cells and only a partial ablation in S. Histological analysis confirmed that no TM cells survived in F while the extracellular matrix remained. The viability assay showed very low PI and no calcein staining in F in contrast to numerous PI-labeled dead TM cells and calcein-labeled viable TM cells in S.

Conclusion: We developed a rapid TM ablation method that uses cyclic freezing that is free of biological or chemical agents and able to produce a decellularized TM scaffold with preserved TM extracellular matrix in an organotypic perfusion culture.

February 4, 2017
Pseudomonas syringae pv. actinidiae (PSA) strain is a major problem for the kiwifruitindustry worldwide. So far, 5 biovars of PSA have been identified, of which the most virulent form is biovar 3. This is the only biovar that has been detected in Chile, which is the third kiwifruit exporter country and is currently suffering from canker disease produced by PSA. Single nucleotide polymorphisms (SNPs) analyses have classified the biovar 3 strain into three groups: the European, the New Zealander and the Chilean groups, which have evolved from a common Chinese PSA ancestor. Although Chilean strains have been used in phylogenetic analysis, there is no information about genomic diversity within this group or whether they present microbiological characteristics that could affect its virulence. In this work we studied 15 Chilean bacterial isolates collected from orchards with canker disease symptoms, and classified them as PSA using a different PCR techniques. To gain more information on the relationship between the isolates we sequenced part of three conserved genes widely used to classify bacterial strains: gtl (Citrate Synthase), rpoD (Sigma subunit of RNA polymerase II) and gyrB (Gyrase B). Using these sequences we performed a phylogenetic analysis that included some PSA reference sequences. Fourteen PSA Chilean isolates were grouped with PSA reference strains and three of them formed a subgroup within the PSA clade, suggesting clear differences at the genomic level among the isolates. We evaluated three microbiological traits in all the isolates: motility (swimming and swarming), and ability to induce a hypersensitive response in tobacco plants. All the isolates were able to induce the hypersensitive response in tobacco plants and were also able to perform both types of movements in appropriated growing conditions.
January 30, 2017

Background. The phytohormone jasmonates (JAs) regulate fundamental plant processes; such as the anthocyanin accumulation during ripening of strawberry, a non-climacteric fruit model. Jasmonoyl-isoleucine (JA-Ile), one of the bioactive JA molecules, mediates binding of the JAZ repressor protein to COI1, an F-box protein forming the SCFCOI1 ubiquitin E3 ligase complex, in Arabidopsis. The COI1-JA-Ile-JAZ complex initiates the JA-signaling pathway leading to early jasmonate responses. Most of Arabidopsis JAZs contain a degron sequence at the Jas domain responsible for interaction with COI1 and JA-Ile. The woodland strawberry (Fragaria vesca) is a model plant for the Rosaceae family, in which the JA-signaling pathway is poorly understood at the molecular level. The aim of this work was to understand the molecular basis of the interaction between the F. vesca COI1 (FvCOI1) and JAZ1 (FvJAZ1) or JAZ8 (FvJAZ8) mediated by JA-Ile.

Methods. Multiple alignments of amino acid sequences and phylogenetic analyses were performed for FvCOI1 and FvJAZ1/8 and their ortholog sequences. The FvCOI1 and FvJAZ1/8 3D structures were built by homology modeling methods, which were further refined and validated by molecular dynamics simulation (MDS). A molecular docking approach along with MDS analysis were used to understand the interaction capacity between a putative degron-like present in FvJAZ1 and FvJAZ8 with the FvCOI1-JA-Ile and FvCOI1-JA complexes.

Results. FvCOI1 and FvJAZ1/8 showed high and moderate identity, respectively, with the corresponding ortholog proteins from other plant species including apple, grape, tomato and Arabidopsis. The resulting FvCOI1 structural model showed that the F-box and LRR domains were highly similar to that described in ArabidopsisCOI1 (AtCOI1) crystal structure. Unexpectedly, we found that FvJAZ1 has a variant IPMQRK sequence respect to the canonical LPIAR(R/K) degron sequence observed in AtJAZ1. The MDS results showed that the FvCOI1-JA-Ile-FvJAZ1 complex was the most stable among all the analyzed ones, and the IPMQRK peptide of FvJAZ1 interacted directly with FvCOI1 and JA-Ile. In contrast, FvJAZ8 did not show a direct interaction with those two components, as expected from previous experimental results for the ortholog AtJAZ8.

Discussion. The present research provides novel insight into the molecular interactions between key JA-signaling components in the model plant F. vesca. Remarkably, we characterized the IPMQRK sequence present in FvJAZ1, a putative variant of the canonical degron previously described in AtJAZ1. We propose that the FvCOI1-JA-Ile-FvJAZ1 complex is stable, and that the degron-like sequence present in FvJAZ1 interacts in a steady manner with FvCOI1-JA-Ile. Up to now, this is the first structural characterization of molecular interactions that may be occurring between the core components of the JA-Ile perception complex in a fleshy fruit-related species.

January 23, 2017
After the conclusion of World War II, members of the Imperial Japanese Army biological warfare Unit 731 testified to a Soviet military court they conducted a live test deployment of plague-infected fleas in Ningpo, Zhejiang Province, a city south of Shanghai with a population of approximately 300,000. The deployment triggered an outbreak involving 78 cases and 74 fatalities (case fatality rate 95%) that included the death of seven families. Children and young adults aged 11-30 were most severely affected. Despite lack of access to effective medical countermeasures, the Ningpo community exhibited a high degree of social cohesion and resilience in the context of effective public health response.
January 6, 2017
In order to understand the distribution and prevalence of Ommatissus lybicus (Homoptera: Tropiduchidae) as well as analyse their current biographical patterns and predict their future spread, comprehensive and highly sophisticated information on the environmental, climatic, and agricultural practices are essential. The analytical techniques available in modern spatial analysis packages, such as Remote Sensing and Geographical Information Systems, can help detect and model spatial links and correlations between the presence, absence and density of O. lybicus in response to climatic, environmental and human factors. The main objective of this paper is to review remote sensing and geographical information analytical techniques that can be applied in mapping and modelling the habitat and population density of O. lybicus in Oman. An exhaustive search of related literature revealed that there are few studies linking location-based infestation levels of pests like the O. lybicus with climatic, environmental and human practice related variables in the Middle East. Our review also highlights the accumulated knowledge and addresses the gaps in this area of research. Furthermore, it makes recommendations for future studies, and gives suggestions on monitoring and surveillance sites that are necessary in designing both local and regional level integrated pest management (IPM) policing of palm tree and other affected cultivated crops.

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