Coupling ZnIn₂S₄ Nanosheets with MoS₂ Hollow Nanospheres as Visible-Light-Active Bifunctional Photocatalysts for Enhancing H₂ Evolution and RhB Degradation

In this study, Mo-glycerate was used as a precursor to create MoS2 hollow nanospheres (HNS), which were then used for the first time to modify ZnIn2S4 nanosheets to create MoS2 HNS/ZnIn2S4 photocatalysts. The findings demonstrate that MoS2 HNS/ZnIn2S4 heterojunctions exhibited remarkably boosted photocatalytic properties and excellent reusability for both RhB degradation and H2 evolution without the use of Pt as a co-catalyst. Among the heterojunctions, the RhB degradation and H2 evolution efficiencies of the optimized MoS2 HNS/ZnIn2S4-3 wt % composite were almost 5 and 34 times higher than those of ZnIn2S4, respectively. The excellent performance of MoS2 HNS/ZnIn2S4-3 wt % might be attributed to the expansion of the visible-light response range and the accelerated separation efficiency of photo-induced carriers, according to the findings of the optical property tests. Based on the established band gap position and characterization results, a potential mechanism for appealing photocatalytic activity over MoS2 HNS/ZnIn2S4 heterojunctions was also postulated

Protective effect of in vaccination trials^a.

a<p>Protective effect was assessed by comparing the worm burden and EPG between pET-26b-<i>Cs</i>Pmy group and PBS group, as well as pcDNA-<i>Cs</i>Pmy group and pcDNA group, respectively. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033703#s3" target="_blank">Results</a> for analysis represented mean ± S.D., and the recovered worm numbers and EPG in groups were compared by Student's <i>t</i>-test.</p>(*)<p><i>p</i><0.05 and.</p>(**)<p><i>p</i><0.01 (compared to corresponding control).</p

Multiple sequence alignment of deduced amino acid sequence of paramyosin among helminthes.

<p><i>C. s-</i>1 (JQ041818) represents the sequence from our <i>C. sinensis</i> metacercaria cDNA plasmid library. <i>C. s-</i>2 (ABN79674.1) represents the sequence submitted by the laboratory from Korea. <i>Paragonimus westermani</i> (<i>P. w</i>, AAY44740.1), <i>Schistosoma haematobium</i> (<i>S. h</i>, BAF62291.1), <i>Schistosoma japonicum</i> (<i>S. j</i>, AAA81003.1), <i>Schistosoma mansoni</i> (<i>S. m</i>, AAA29915.1), <i>Taenia solium</i> (<i>T. s</i>, AAK58494.1) and <i>Echinococcus granulosus</i> (<i>E. g</i>, CAA79849.1). Amino acids shared among helminthes were indicated in black, high conserved amino acids among helminthes were indicated in gray. B-cell and T-cell linear epitopes were indicated with full lines and dotted lines, respectively.</p

Expression and purification of recombinant pET-26b-<i>Cs</i>Pmy identified by 8% SDS-PAGE.

<p>(A) Expression of pET-26b-<i>Cs</i>Pmy. Protein molecular weight marker (M), lysate of <i>E. coli</i> with pET-26b(+) before induction with IPTG (lane 1) and after induction (lane 2), lysate of <i>E. coli</i> with pET-26b-<i>Cs</i>Pmy before induction with IPTG (lane 3) and after induction (lane 4), supernatant of induced <i>E. coli</i> with pET-26b-<i>Cs</i>Pmy (lane 5) and sediment (lane 6). (B) Denaturation of inclusion bodies containing pET-26b-<i>Cs</i>Pmy. Supernatant collected from inclusion bodies dissolved in 2 M urea (lane 1) and 6 M urea (lane 2). (C) Purification of pET-26b-<i>Cs</i>Pmy. Protein eluted with 40 mM imidazole (lane 1–3), 80 mM imidazole (lane 4–8), 100 mM imidazole (lane 9–12), 200 mM imidazole (lane 13–14). Proteins were visualized by Coomassie Blue staining, the protein bands were around 100 kDa.</p

Immunohistochemical localization of <i>Cs</i>Pmy at adult worm and metacercaria.

<p>Adult worms and metacercariae of <i>C. sinensis</i> were fixed with 4% paraformaldehyde, embedded with paraffin and sliced into 3–5 µm in thick. The sections were blocked with normal goat serum overnight at 4°C, and then incubated with primary antibody (1∶200 dilutions) at room temperature for 2 h. After washing procedures, the sections were incubated with goat anti-rat IgG Alexa Fluor 594 (1∶400 dilutions) at room temperature for 1 h in dark. The images were captured under fluorescence microscope (ZEISS, Goettingen, Germany). Panel A–H, adult worm of <i>C. sinensis</i>. Pane I–L, metacercariae of <i>C. sinensis</i>. Pane A, B, E, F, I and J were sections treated with anti-pET26b-<i>Cs</i>Pmy serum. C, D, G, H, K and L were sections treated with naïve serum and imaged under the same conditions. Specific immunofluorescence was indicated in red (pane A, E and I), while no immunofluorescence was detected in pane C, G and K. Corresponding white light of parasite was panel B, D, F, H, J and L. <b>T</b>, tegument. <b>OS</b>, oral sucker. <b>V</b>, vitellarium. <b>CW</b>, cyst wall. <b>EB</b>, excretory bladder. Magnification for adult worm and metacercaria were ×100 and ×400, respectively.</p

Identification of <i>Cs</i>Pmy by SDS-PAGE and Western blot analysis.

<p>Protein molecular weight marker (M), purified pET-26b-<i>Cs</i>Pmy protein (lane 1), TWE of adult worm (lane 2), TWE of metacercaria (lane 3), TWE of cercaria (lane 4), TWE of egg (lane 5), cyst wall proteins of metacercaria (lane 6), and soluble tegumental components of adult worm (lane 7). (A) 8% SDS-PAGE. (B) Western blot analysis. Corresponding proteins were subjected to 8% SDS-PAGE and immobilized onto the membrane, then the membrane was incubated with anti-pET-26b-<i>Cs</i>Pmy rat serum (1∶2000 dilutions) at room temperature for 2 h. Subsequently, the membrane was followed by incubation with rabbit anti-rat IgG HRP-conjugated secondary antibody (1∶2000 dilutions) at room temperature for 1 h. 2 µg of purified pET-26b-<i>Cs</i>Pmy protein and 10 µg of TWE were loaded per lane. SDS-PAGE was visualized by Coomassie Blues staining and the protein bands that might be native paramyosin in different life stages were indicated with arrows. Western blot was visualized by ECL method, the detected protein bands were around 100 kDa.</p

IgG isotype induced by <i>Cs</i>Pmy measured by ELISA.

<p>1 µg/well recombinant pET-26b-<i>Cs</i>Pmy protein was coated on the plates and blocked with 5% skimmed milk. Immune sera from week 2 to week 6 were diluted at 1∶400. Rat sera immunized with PBS and pcDNA were measured under the same conditions as negative controls. IgG (1∶20000 dilutions), IgG1 and IgG2a (1∶1000 dilutions) were used as secondary antibodies. (A) Immune responses induced by pET-26b-<i>Cs</i>Pmy. (B) Immune responses induced by pcDNA-<i>Cs</i>Pmy.</p

Transcriptional level of <i>Cs</i>Pmy at different developmental stages of <i>C. sinensis</i> by qRT-PCR experiments.

<p>Total RNA from four stages (adult worm, metacercaria, cercaria and egg) were extracted by TRIzol methods and spectrophotometrically quantitated. Reverse transcription reactions were carried out to get the first-strand cDNA with the same quantity of total RNA as the template (1 µg). β-actin of <i>C. sinensis</i> (accession number: EU109284) was used as the transcription control. The real-time PCR amplification was performed using the LightCycler480 instrument (Roche, Switzerland) using the SYBR Premix ExTaq Kit. The LightCycler480 software (version 1.5) was used to analyze the data according to the 2^−ΔΔCt method <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033703#pone.0033703-Livak1" target="_blank">[27]</a>. The amplification of egg was employed as the calibrator to evaluate relative expression levels of <i>Cs</i>Pmy.</p

Proteins from cyst wall of <i>C. sinensis</i> metacercariae identified by HPLC-MS/MS.

<p>Prot_acc, protein accession number in databases; Prot description, the most likely matched protein name in the NCBI database. Prot_score, the similarity that peptide matched in the NCBI database. prot_match, the matched peptides in the NCBI database. Prot_cover, the percent of identified amino acids in target amino acid sequence. Prot_mass, molecular mass of matched proteins. Score >50 indicates extensive similarity (<i>p</i><0.05) in the NCBI database.</p

Low Divergence of <i>Clonorchis sinensis</i> in China Based on Multilocus Analysis

<div><p><i>Clonorchis sinensis</i>, an ancient parasite that infects a number of piscivorous mammals, attracts significant public health interest due to zoonotic exposure risks in Asia. The available studies are insufficient to reflect the prevalence, geographic distribution, and intraspecific genetic diversity of <i>C. sinensis</i> in endemic areas. Here, a multilocus analysis based on eight genes (ITS1, <i>act</i>, <i>tub</i>, <i>ef-1a</i>, <i>cox1</i>, <i>cox3</i>, <i>nad4</i> and <i>nad5</i> [4.986 kb]) was employed to explore the intra-species genetic construction of <i>C. sinensis</i> in China. Two hundred and fifty-six <i>C. sinensis</i> isolates were obtained from environmental reservoirs from 17 provinces of China. A total of 254 recognized Multilocus Types (MSTs) showed high diversity among these isolates using multilocus analysis. The comparison analysis of nuclear and mitochondrial phylogeny supports separate clusters in a nuclear dendrogram. Genetic differentiation analysis of three clusters (A, B, and C) showed low divergence within populations. Most isolates from clusters B and C are geographically limited to central China, while cluster A is extraordinarily genetically diverse. Further genetic analyses between different geographic distributions, water bodies and hosts support the low population divergence. The latter haplotype analyses were consistent with the phylogenetic and genetic differentiation results. A recombination network based on concatenated sequences showed a concentrated linkage recombination population in <i>cox1</i>, <i>cox3, nad4</i> and <i>nad5</i>, with spatial structuring in ITS1. Coupled with the history record and archaeological evidence of <i>C. sinensis</i> infection in mummified desiccated feces, these data point to an ancient origin of <i>C. sinensis</i> in China. In conclusion, we present a likely phylogenetic structure of the <i>C. sinensis</i> population in mainland China, highlighting its possible tendency for biogeographic expansion. Meanwhile, ITS1 was found to be an effective marker for tracking <i>C. sinensis</i> infection worldwide. Thus, the present study improves our understanding of the global epidemiology and evolution of <i>C. sinensis</i>.</p></div