Heterologous expression of pediocin/papA in Bacillus subtilis.

Listeria monocytogenes is a food-borne pathogen. Pediocin is a group IIα bacteriocin with anti-listeria activity that is naturally produced by Pediococcus acidilactic and Lactobacillus plantarum. The pedA/papA gene encodes pediocin/plantaricin. In native hosts, the expression and secretion of active PedA/PapA protein rely on the accessory protein PedC/PapC and ABC transporter PedD/PapD on the same operon. The excretion machines were also necessary for pediocin protein expression in heterologous hosts of E. coli, Lactobacillus lactis, and Corynebacterium glutamicum. In this study, two vectors carrying the codon sequence of the mature PapA peptide were constructed, one with and one without a His tag. Both fragments were inserted into the plasmid pHT43 and transformed into Bacillus subtilis WB800N. The strains were induced with IPTG to secrete the fused proteins PA1 and PA2. Supernatants from both recombinant strains can inhibit Listeria monocytogenes ATCC54003 directly. The fused protein possesses inhibition activity as a whole dispense with removal of the leading peptide. This is the first report of active pediocin/PapA expression without the assistance of PedCD/PapCD in heterogeneous hosts. In addition, the PA1 protein can be purified by nickel-nitrilotriacetic acid (Ni-NTA) metal affinity chromatography.

Isolation, Structural Characterization and Antidiabetic Activity of New Diketopiperazine Alkaloids from Mangrove Endophytic Fungus Aspergillus sp. 16-5c.

Six new DIKETOPIPERAZINE alkaloids aspergiamides A-F (1-6), together with ten known alkaloids (7-16), were isolated from the mangrove endophytic fungus Aspergillus sp. 16-5c. The structures of the new compounds were elucidated based on 1D/2D NMR spectroscopic and HR-ESIMS data analyses. The absolute configurations of aspergiamides A-F were established based on the experimental and calculated ECD data. All the compounds were evaluated for the antidiabetic activity against α-glucosidase and PTP1B enzyme. The bioassay results disclosed compounds 1 and 9 exhibited significant α-glucosidase inhibitory with IC50 values of 18.2 and 7.6 µM, respectively; compounds 3, 10, 11, and 15 exhibited moderate α-glucosidase inhibition with IC50 values ranging from 40.7 to 83.9 µM; while no compounds showed obvious PTP1B enzyme inhibition activity.

Isobenzofuranone monomer and dimer derivatives from the mangrove endophytic fungus Epicoccum nigrum SCNU-F0002 possess α-glucosidase inhibitory and antioxidant activity.

Four new isobenzofuranone monomers, (+)-epicoccone C ((+)-1), (-)-epicoccone C ((-)-1), epicoccone D (2), epicoccone E (3) and one new isobenzofuranone dimer, epicolactone A (4), together with four known related dimers were obtained from the fermentation of an endophytic fungus, Epicoccum nigrum SCNU-F0002, which was isolated from the fresh fruit of the mangrove plant Acanthus ilicifolius L. Their structures were elucidated on the basis of spectroscopic methods, single-crystal X-ray diffraction analysis, and electronic circular dichroism calculations. These isolated compounds (1-8) were evaluated for their antioxidant activity and α-glucosidase enzyme inhibitory activity. All of the compounds except 5 exhibited more potent α-glucosidase inhibitory effect than acarbose. Most of the compounds showed superior antioxidant activity with IC50 values ranging from 10.2 to 15.3 µM than positive control, gallic acid and vitamin C.

Immunogenic potential and protective efficacy of a sptP deletion mutant of Salmonella Enteritidis as a live vaccine for chickens against a lethal challenge.

Salmonella Enteritidis (SE) is a highly adapted pathogen causing severe economic losses in the poultry industry worldwide. Chickens infected by SE are a major source of human food poisoning. Vaccination is an effective approach to control SE infections. This study evaluated the immunogenicity and protective efficacy of a SE sptP deletion mutant (C50336ΔsptP) as a live attenuated vaccine (LAV) candidate in chickens. 14 day-old specific pathogen-free (SPF) chickens were intramuscularly immunized with various doses of C50336ΔsptP. Several groups of chickens were challenged with the virulent wild-type SE strain Z-11 via the same route at 14 days post vaccination. Compared to the control group, the groups vaccinated with 1 × 106, 1 × 107 and 1 × 108 colony-forming units (CFU) of C50336ΔsptP exhibited no clinical symptoms after immunization. Only slight pathological changes occurred in the organs of the 1 × 109 CFU vaccinated group. C50336ΔsptP bacteria were cleared from the organs of immunized chickens within 14 days after vaccination. Lymphocyte proliferation and serum cytokine analyses indicated that significant cellular immune responses were induced after the vaccination of C50336ΔsptP. Compared to the control group, specific IgG antibody levels increased significantly in vaccinated chickens, and the levels increased markedly after the challenge. The 1 × 107, 1 × 108, and 1 × 109 CFU vaccinated chickens groups showed no clinical symptoms or pathological changes, and no death after the lethal challenge. Whereas severe clinical signs of disease and pathological changes were observed in the control group chickens after the challenge. These results suggest that a single dose of C50336ΔsptP could be an effective LAV candidate to against SE infection in chickens.

Identification and development of microsatellite (SSRs) makers of Exbucklandia (HAMAMELIDACEAE) by high-throughput sequencing.

Hamamelidaceae (Saxifragales, previously Rosales) comprises approximately six subfamily, 30 genera and 140 species, most of which are Tertiary relicts. Exbucklandia is the only genus of the subfamily Exbucklandioideae, Hamelidaceae, containing only 2-4 species. Of them, the species E. longipetala H. T. Chang is endemic to China and listed as endangered in The Biodiversity Red List of China: Higher Plant, yet some taxonomists put forward that E. longipetala should be merged into E. tonkinensis (Lecomte) H. T. Chang. Currently, there was nearly no phylogeographic studies on this genus possibly due to the deficiency of efficient molecular markers. In this study, we sequenced the genome of E. tonkinensis based on high throughput sequencing technology, and obtained approximately 6 G raw data, which was further de novo assembled into 303,481 contigs. Based on them, 15,326 SSRs were identified from 13,596 contigs, and primers were successfully designed for 10,660 SSRs. A total of 139 paired primers were synthesized, 106 of them were successfully amplified in six Exbucklandia individuals with expected PCR product size, and 24 demonstrated to be polymorphic among three Exbucklandia populations. Accordingly, the expected and observed heterozygosity were between 0.097-0.717 and 0.098-0.583. Based on these efforts, future researches on genetic diversity and population structure of Exbucklandia can be performed to understand its phylogenetic origins and phylogeographic pattern.

The Purification, Characterization, and Biological Activity of New Polyketides from Mangrove-Derived Endophytic Fungus Epicoccum nigrum SCNU-F0002.

Six new polyketides, including one coumarin (1), two isocoumarins (2 and 3), dihydroradicinin (4), and two benzofuranone derivatives (7 and 8), together with seven known analogues (5-6 and 9-13) were isolated from the culture of the mangrove endophytic fungus Epicoccum nigrum SCNU-F0002. The structures were elucidated on the interpretation of spectroscopic data. The absolute configuration of Compounds 2 and 3 were determined by comparison of their ECD spectra with the data of their analogue dihydroisocoumarins described in the literature. The absolute configuration of 4 was determined by single-crystal X-ray diffraction. All the compounds were screened for their antioxidant, antibacterial, anti-phytopathogenic fungi and cytotoxic activities. Using a DPPH radical-scavenging assay, Compounds 10-13 showed potent antioxidant activity with IC50 values of 13.6, 12.1, 18.1, and 11.7 µg/mL, respectively. In addition, Compounds 6 and 7 showed antibacterial effects against Bacillus subtilis (ATCC 6538), Escherichia coli (ATCC 8739), and Staphylococcus aureus (ATCC 6538), with MIC values in the range of 25-50 µg/mL.

A polarizable dipole-dipole interaction model for evaluation of the interaction energies for N-H···O=C and C-H···O=C hydrogen-bonded complexes.

In this article, a polarizable dipole-dipole interaction model is established to estimate the equilibrium hydrogen bond distances and the interaction energies for hydrogen-bonded complexes containing peptide amides and nucleic acid bases. We regard the chemical bonds N-H, C=O, and C-H as bond dipoles. The magnitude of the bond dipole moment varies according to its environment. We apply this polarizable dipole-dipole interaction model to a series of hydrogen-bonded complexes containing the N-H···O=C and C-H···O=C hydrogen bonds, such as simple amide-amide dimers, base-base dimers, peptide-base dimers, and ß-sheet models. We find that a simple two-term function, only containing the permanent dipole-dipole interactions and the van der Waals interactions, can produce the equilibrium hydrogen bond distances compared favorably with those produced by the MP2/6-31G(d) method, whereas the high-quality counterpoise-corrected (CP-corrected) MP2/aug-cc-pVTZ interaction energies for the hydrogen-bonded complexes can be well-reproduced by a four-term function which involves the permanent dipole-dipole interactions, the van der Waals interactions, the polarization contributions, and a corrected term. Based on the calculation results obtained from this polarizable dipole-dipole interaction model, the natures of the hydrogen bonding interactions in these hydrogen-bonded complexes are further discussed.

Landform and lithospheric development contribute to the assembly of mountain floras in China.

Although it is well documented that mountains tend to exhibit high biodiversity, how geological processes affect the assemblage of montane floras is a matter of ongoing research. Here, we explore landform-specific differences among montane floras based on a dataset comprising 17,576 angiosperm species representing 140 Chinese mountain floras, which we define as the collection of all angiosperm species growing on a specific mountain. Our results show that igneous bedrock (granitic and karst-granitic landforms) is correlated with higher species richness and phylogenetic overdispersion, while the opposite is true for sedimentary bedrock (karst, Danxia, and desert landforms), which is correlated with phylogenetic clustering. Furthermore, we show that landform type was the primary determinant of the assembly of evolutionarily older species within floras, while climate was a greater determinant for younger species. Our study indicates that landform type not only affects montane species richness, but also contributes to the composition of montane floras. To explain the assembly and differentiation of mountain floras, we propose the 'floristic geo-lithology hypothesis', which highlights the role of bedrock and landform processes in montane floristic assembly and provides insights for future research on speciation, migration, and biodiversity in montane regions.

Effect of repetitive transcranial magnetic stimulation combined with transcranial direct current stimulation on post-stroke dysmnesia: A preliminary study.

OBJECTIVE: This study examined the effect of repetitive transcranial magnetic stimulation (rTMS) combined with transcranial direct current stimulation (tDCS) as a bimodal neuromodulatory approach for post-stroke dysmnesia. METHODS: Thirty-four patients with post-stroke dysmnesia were randomly allocated into a sham group treated with neither rTMS nor tDCS, a group treated with rTMS, and a group treated with a combination of rTMS and tDCS. All three groups received cognitive rehabilitation training for 4 weeks. The memory function of each group before and after the intervention was assessed using the Montreal Cognitive Assessment (MoCA) and Rivermead Behavioral Memory Test (RBMT) scales, as well as in terms of the Mismatch Negativity（MMN）and P300 of event-related potentials. RESULTS: The sham, rTMS, and rTMS-tDCS groups all showed improvement in the total MoCA score after the intervention. Delayed recall, a MoCA item, scored better in the rTMS-tDCS group than in the rTMS and sham groups. Delayed processing, an RBMT item, scored better in the rTMS-tDCS combination group than in the rTMS and sham groups. MMN and P300 latency was significantly shorter in the rTMS-tDCS combination group. CONCLUSION: rTMS-tDCS bimodal stimulation was more effective than cognitive rehabilitation or rTMS alone in treating patients with post-stroke dysmnesia, offering new possibilities for enhancing cognitive function and treating post-stroke dysmnesia.

Highly pH-stable lanthanide MOFs: a tunable luminescence and ratiometric luminescent probe for sulfamethazine.

By selecting a bisphosphonic ligand H4L (H4L = 4-F-C6H4CH2N(CH2PO3H2)2) and a coligand oxalate (H2C2O4), three isostructural lanthanide metal-organic frameworks (Ln-MOFs) with a 2D layer structure, [Ln(H3L)(C2O4)]·2H2O (Ln = Eu (1), Gd (2), or Tb (3)), were hydrothermally prepared. By tuning the molar ratio of Eu3+, Gd3+, and Tb3+ in the above reactions, six bimetallic or trimetallic doped Ln-MOFs (EuxTb1-x (x = 0.02 (4), 0.04 (5), and 0.06 (6)), Gd0.94Eu0.06 (7), Gd0.96Tb0.04 (8) and Gd0.95Tb0.03Eu0.02 (9)) were obtained. The powder X-ray diffraction (PXRD) patterns of doped Ln-MOFs 4-9 show that they are isomorphous with 1-3. The bimetallic doped Ln-MOFs show a gradual variation of luminous colors between yellow-green, yellow, orange, pink, and light blue. Meanwhile, the trimetallic doped Gd0.95Tb0.03Eu0.02 Ln-MOF (9) displays near white-light emission with a quantum yield of 11.39%. Interestingly, the luminous inks of 1-9 are invisible and color tunable, which makes it possible to promote their anti-counterfeiting applications. Additionally, 3 displays good thermal, water, and pH stabilities, which provides the feasibility for its sensing application. The luminescence sensing experiments show that 3 can serve as a highly selective, reusable, and ratiometric luminescent sensor of sulfamethazine (SMZ). Moreover, 3 shows an excellent SMZ detection performance in real samples, such as mariculture water and real urine. Owing to the visible variation of the response signal under a UV lamp, portable SMZ test paper was prepared.

Molecular phylogenetic analyses based on the complete plastid genomes and nuclear sequences reveal Daphne (Thymelaeaceae) to be non-monophyletic as current circumscription.

The diverse members of the genus Daphne are prized for their fragrant flowers. Despite being promising ornamental plants in many countries, genetic information of Daphne is scarce. In this study, the plastomes of four species and one variety of Daphne were sequenced and analyzed. The plastomes were typical and contained a pair of inverted repeat (IR) regions that separated the large single-copy (LSC) region from the small single-copy (SSC) region. With a length ranging from 132,869 bp (D. genkwa) to 174,773 bp (D. championii), 106 to 141 genes were predicted. Comparative plastome analysis of the newly sequenced plastomes with four publicly available Daphne plastomes identified an expansion of the IRs, sequence variations, and mutational hotspots. Phylogenetic analyses indicated that the genus Daphne in its current circumscription is polyphyletic. Daphne genkwa was nested within the genus Wikstroemia, while D. championii was well resolved as sister to Edgeworthia. These findings concurred with results from our study that used nuclear ribosomal internal transcribed spacer sequence data. The conflicts on the molecular placement of D. championii and D. genkwa and the present taxonomic classification in Daphne suggest that a new intergeneric classification system of Daphneae warrants consideration.

Salmonella Pullorum spiC mutant is a desirable LASV candidate with proper virulence, high immune protection and easy-to-use oral administration.

Live attenuated Salmonellavaccine (LASV) is considered to be an effective contributory measure during the control of Salmonella infection. A Salmonella Pullorum spiC mutant was evaluated comprehensively as a LASV candidate (LASV-p) for broilers in terms of safety and immunogenicity. LASV-p was adminstered to 3-day broilers by intramuscular injection. The LD50 increased 126 fold, and no tissue lesions were observed in the liver, spleen and cecum, in comparison with the control group inoculated with PBS and a passive group by wild-type Salmonella. Growth rates of all broilers were normal and not affected. LASV-p persisted in vivo until 21 days in liver, 28 days in spleen and 35 days in feces, and induced high levels of humoral IgG and mucosal IgA. Cellular immunity was also stimulated in the form of antigen-specific lymphocyte proliferation and higher counts of CD3+CD8+ T cells and increased expression of mRNA of Th1 cytokines, IFN-γ and IL-2, in the early stage, and Th2 cytokines, IL-4 and IL-10, in the later stages. LASV-p provided at least 90% immuneprotection against a wild-type Salmonella Pullorum and cross-protection in different degree against other Salmonella searovars. Oral vaccine could also offer high immune protection of 87.5%. These results indicated that LASV-p vaccine candidate had a high level of safety and immune protection and it might be developed as a novel easy-to-use oral vaccine to improve poultry health in the future.

Two new secondary metabolites from the mangrove endophytic fungus Pleosporales sp. SK7.

Two new compounds, an abscisic acid-type sesquiterpene (1), and one asterric acid derivative (2), together with three known compounds (3-5) were isolated from mangrove endophytic fungus Pleosporales sp. SK7. The structures of these metabolites were determined by NMR, X-ray crystal diffraction, CD and HR-ESI-MS. All compounds were tested for their antibacterial, antioxidant and cytotoxic activities, among these compounds, 5 showed cytotoxicity against MDA-MB-435 cell with an IC50 of 25.96 ± 0.32 µM.

New furo[3,2-h]isochroman from the mangrove endophytic fungus Aspergillus sp. 085242.

Four new compounds, asperisocoumarin G (1), asperisocoumarin H (2), (±)-asperisocoumarin I [(±)-3], along with the known pergillin (4) and penicisochroman L (5) were isolated from a mangrove endophytic fungus Aspergillus sp. 085242 by further chemical investigation. The structures of the new compounds, including their absolute configurations, were established by analysis of HR-ESI-MS and NMR spectroscopic data, and ECD calculation. Asperisocoumarins G-I (1-3) were new isocoumarins belonging to the class of furo[3, 2-h]isocoumarins which are rarely found in natural sources. All of the isolated compounds were evaluated for their α-glucosidase inhibitory effects, and compounds 1 and 4 showed moderate α-glucosidase inhibitory activity, respectively. In an antimicrobial test, the racemate of 3 showed antibacterial activity against Salmonella.

A new polyoxometalate-templated Mo/V-oxide-based organic-inorganic hybrid framework with a honeycomb-like structure.

A new polyoxometalate-templated Mo/V-oxide-based organic-inorganic hybrid framework with a honeycomb-like structure has been hydrothermally synthesized and its electrocatalytic property was investigated.

Femtosecond Optical Annealing Induced Polymer Melting and Formation of Solid Droplets.

Interaction between femtosecond laser pulses with polymeric thin films induced transient optical annealing of the polymer molecules. Melting of the polymer films took place during the transient annealing process, so that a solid-liquid-solid phase transition process was observed. Ultrafast cooling of the melting polymer produced solidified droplets. Microscopic and spectroscopic characterization revealed that the polymer molecules were rearranged with preferable H-aggregation to reach the lowest formation energy during the melting process. Intermolecular coupling was enhanced due to the modified molecular arrangement. This observation of melting of polymeric semiconductors due to the interaction with femtosecond light pulses is potentially important for better understanding laser-matter interactions and for exploring organic optoelectronic devices through special material processing.

Thiocladospolide E and cladospamide A, novel 12-membered macrolide and macrolide lactam from mangrove endophytic fungus Cladosporium sp. SCNU-F0001.

Chemical investigation of the mangrove endophytic fungus Cladosporium sp. SCNU-F0001 resulted in the isolation and identification of a new macrolide compound named thiocladospolide E (1) and a novel macrolide lactam named cladospamide A (2), along with the known cladospolide B (3). The structures were elucidated based on spectroscopic methods, and the absolute configurations were determined by X-ray diffraction and HPLC analysis after chemical derivatization. All compounds were tested for their antibacterial and cytotoxic activity.

Induction of arthritis in chickens by infection with novel virulent Salmonella Pullorum strains.

Salmonella enterica subsp. enterica serovar Gallinarum biovar Pullorum (Salmonella Pullorum) is a host-specific serovar causing systemic infection with high mortality in young chicks. Pullorum disease is characterized by white diarrhea. However, arthritis has become increasingly frequent recently, particularly in southern China. The aim of the present study was to determine the pathogenesis and arthritis induction of new Salmonella Pullorum isolates. We isolated and identified five Salmonella Pullorum strains from broilers with bacterial arthritis and lameness in a commercial poultry farm. Four of five isolates were resistant to at least three classes of antibiotics and were defined as multidrug-resistant Salmonella Pullorum. All isolates had the same CRISPR sequence type and belonged to a single major cluster. The isolates exhibited high capability of biofilm formation, which may facilitate their dispersal and survival in hostile habitats, and showed high virulence based on embryo lethality and inoculation of newly hatched chicks. Tissue distribution analysis confirmed that SP1621 was more adapted to colonize the joint when compared to the white diarrhoea-causing Salmonella Pullorum reference strain S06004. Reproducible arthritis and typical joint lesions were observed in SP1621-infected chicks, and histopathological examination showed necrotic synovitis and cartilage tissue hyperplasia of the joint. Koch's postulates were confirmed when the novel Salmonella Pullorum strain was re-isolated from the joint tissues of experimentally inoculated chicks. These novel Salmonella Pullorum isolates have unique ability to induce arthritis in chickens, representing expanded pathogenic diversity in China. These results suggest the need for strict control strategies and new vaccines to prevent the disease.

2-(1,3-Dithian-2-yl-idene)-1-phenyl-butane-1,3-dione.

The title compound, C(13)H(12)O(2)S(2), belonging to the group of dioxoketene cyclic S,S-acetals, was prepared from the corresponding dione in high yield. In the structure, the C=O and C=C bonds are not coplanar, with O=C-C=C torsion angles of -36.8 (4) and -21.0 (4)°. The dithian ring has a twisted conformation.

Transient localized surface plasmon induced by femtosecond interband excitation in gold nanoparticles.

Localized surface plasmon resonance (LSPR) is essentially a collective oscillation of free electrons in nanostructured metals. Interband excitation may also produce conduction-band electrons above the Fermi level. However, a question here is whether these excited electrons can take part in plasmonic oscillation. To answer this question, femtosecond pump-probe measurements on gold nanoparticles were performed using interband excitation, where the pump pulse produced a large amount of electrons in the sp-conduction band and left holes in the d-band. Probing by transient absorption spectroscopy, we resolved an induced LSPR feature located at a red-shifted spectrum. This feature cannot be observed for a pumping photon energy lower than the threshold for interband transition. The commonly observed red-shift or broadening of LSPR spectrum due to electron-electron and electron-phonon scattering under strong optical excitation can be ruled out for understanding this feature by a comparison between the plasmonic dynamics at a pump above and below the interband-transition threshold. In particular, a "holding" time of about 1 ps was resolved for the interband-excitation-induced electrons to relax to the LSPR oscillation.