Electrospun Mussel-derived Silk Fibers.

BACKGROUND: Though there are many patents on silk, patents on sea silk are rare. Sea silk is one of the most coveted materials in the world, and the technology to make sea silk is at an extremely high risk of extinction. Unlike spider dragline silk and silkworm silk, this natural silk has been forgotten in the academic commune for millennia, though it has many fascinating properties: high strength, remarkable adhesion, extreme lightweight, and others. METHODS: Here we report that mussel-derived silk fibers can be fabricated by electrospinning. Instead of extracting proteins from byssus, we directly use the protein solution from alive blue mussels, which are intensely commercially used. The protein solution and the polyvinyl alcohol solution are mixed together to produce mussel-based silk fibers. RESULTS: The mussel-based silk fibers have many special properties like high mechanical strength, remarkable super-contraction and good wetting properties. CONCLUSION: The electrospinning mussel-based silk fibers have the potential for use as a replacement for the rarest sea silk and as a new bio-inspired material with multi-functions.

[Planktonic Algae's Distribution and Correlation with Dissolved Organic Matters' Fluorescence in the End of the Three Gorges Reservoir's Back Water Zone].

For researching the community structure, composition, and distribution of the planktonic algae in the Three Gorges Reservoir (TRG), especially, within TRG water levels' fluctuating, mpacting to water quality with the algae's growing, 6 sampling cites in the end of the TGR's back water zone, from Chaotianmen to Taihonggang Town, are chosen to monitor, at the sensitive stages time of algal bloom between March to May 2012, namely, water levels lowing from the highest impounding. The community's structure, composition, and distribution of the planktonic algae in the TGR, and the correlation of water quality parameters and DOM's fluorescence features with algae density, are obtained. According to the experimental results, blue algae fibre, chlorella, melosira, navicula are the dominated algae in the end of the TGR's back water zone, from Chaotianmen to Taihonggang Town, algae density are 0.40~0.56 × 10(6), 1.9~0.8 × 10(6), 0.36~0.25 × 10(6), 0.42~0.15 × 10(6) cells · L(-1) respectively. Besides, anabaena, phormidium, cladophora, feather, ovate algae are existing in only limited 2 sections. The fitting results reveal obvious linear correlation of the EEM characteristics of DOM with the 4 kinds of dominated algal density, which could be useful conference for the algae bloom monitor, conveniently and effectively.

Improved oral bioavailability of breviscapine via a Pluronic P85-modified liposomal delivery system.

OBJECTIVES: Breviscapine, a hydrophobic drug used for treating cardiovascular disease, was encapsulated in liposomes to improve its pharmaceutical characteristics. This study describes a novel liposome composition approach to specifically inhibit the P-glycoprotein efflux system. METHODS: Breviscapine-loaded Pluronic P85-coated liposomes were prepared by the thin film hydration technique. The particle size, zeta potential and encapsulation efficiency of the formulations were characterized. In-vitro drug release and permeability of Caco-2 cells were investigated. In-vitro characteristics and pharmacokinetics of the liposomes were evaluated in rat studies. KEY FINDINGS: The Pluronic P85-modified liposomes dispersed individually and had an approximate diameter of 118.8 ± 4.9 nm and a zeta potential of -35.4 ± 1.5 mV. Encapsulation efficiency was more than 90%. The use of the P85-coated liposomes resulted in significantly (P<0.05) increased absorption of breviscapine in Caco-2 cells and in 5.6-fold enhancement in its oral bioavailability in rats. CONCLUSION: The P85-modified liposomes for the oral delivery of breviscapine were prepared using l-α-phosphatidylcholine (soy-hydrogenated) and cholesterol with a narrow size distribution. This method seems to effectively enhance the bioavailability of breviscapine in rats.

[Research on the degradation of BaP with potassium ferrate characterized by fluorescence].

The degradation of Benzo(a)pyrene (BaP) by potassium ferrate was researched by means of multiple fluorescence spectroscopic methods such as synchronous, time-scan, excitation emission matrix (EEM) and photometry, under the optimal condition. Within the degradation process, the characteristics of the BaP's concentration at different time-intervals, and the kinetics of the degradation of BaP by potassium ferrate were discussed. From the experimental data, both synchronous and EEM spectra's results showed that the concentration of BaP was reduced 90% by potassium ferrate within 20 s after degradation, and the reaction process was very slow after 60 s. The degradation kinetic equation, ln(F0/Ft) = 0.563 2t + 0.171 2, (R2 = 0.994 2), was obtained through a convenient and fast way combining the time-scan fluorescence data and photometry data, and the photometry included the synchronous photometry and emission photometry. According to the kinetic equation, the degradation of BaP by potassium ferrate was in accord with the order of the first order reaction. So this article could provide a very useful conference for the research on the pollutant degradation by potassium ferrate, especially for the degradation process and the degradation mechanisms.

[Simultaneous determination of heavy metals in waste and biogas slurry of scale farm by microwave digestion/inductively coupled plasma-mass spectrometry].

A new microwave digestion/inductively coupled plasma mass spectrometry (ICP-MS) for simultaneous determination of 8 heavy metals including Zn, Cu, As, Pb, Cd, Cr, Hg and Ni in wastes of scale hoggery from livestock parks in Chongqing and its biogas slurry after anaerobic digestion, with which an all round, fast analysis of multiple heavy metals in biogas slurry was realized. From the experiment, Zn, Cu, As, Pb, Cd, Cr, Hg, and Ni were the dominating heavy metals in biogas slurry. The linear correlation coefficients for the heavy metals were between 0.999 89 and 0.999 98, indicating a good linear correlation. The determination limits were between 0.79 and 25.0 ng x L(-1), recovery adding standard between 96.5% and 107.6%, RSD between 1.06% and 4.35%. These parameters revealed that the method has good precision and accuracy, which could be applied to the heavy metals detection in biogas slurry, and provide reference for its post-use.

[Combined application of multiple fluorescence in research on the degradation of fluoranthene by potassium ferrate].

The degradation of fluoranthene was researched by combined means of multiple fluorescence spectra, including emission, synchronous, excitation emission matrix (EEM), time-scan and photometry. The characteristics of the degradation and fluoranthene molecular changes within the degradation's process were also discussed according to the information about the degradation provided by all of the fluorescence spectra mentioned above. The equations of fluoranthene's degradation by potassium ferrate were obtained on the bases of fitting time-scan fluorescence curves at different time, and the degradation's kinetic was speculated accordingly. From the experimental results, multiple fluorescence data commonly reflected that it had same degradation rate at the same reaction time. t = 10 s, and the degradation rate is -55%, t = 25 s, -81%, t = 40 s, -91%. No new fluorescent characteristic was observed within every degradation' stage. The reaction stage during t < or = 20 s was crucial, in which the degradation process is closest to linear relationship. After this beginning stage, the linear relationship deviated gradually with the development of the degradation process. The degradation of fluoranthene by potassium ferrate was nearly in accord with the order of the first order reaction.

[Function of luxS gene in sulfurmetabolism of Streptococcus mutans].

OBJECTIVE: To investigate the function of luxS in sulfurmetabolism of Streptococcus mutans (Sm). METHODS: The growth with absorbency (A) of the standards and mutant strains was measured and analyzed in the sulfur-limited defined medium at different periods. The laser scanning confocal microscopy (LSCM) was used to observe and compare the biofilm thickness of the two kinds of strains at different culture conditions. RESULTS: The significant increases in the thickness of mutant strain biofilm and its growth were observed after the addition of cysteine, but did not reach the standards strain levels (P < 0.05). The growth and the biofilm thickness of the mutant strains were (1.301 ± 0.009) and (45.009 ± 0.429) µm. When methionine and S-adenosylhomocysteine of certain concentrations were respectively added, the biofilm thickness and the growth of mutant strain were raised but did not reach the level of the standards strain at 24 h (P < 0.05), but at 48 h they did. When the methionine was added in the mutant strains for 24 h, the biofilm thickness and the growth of mutant strain were (0.448 ± 0.028) and (37.068 ± 2.392) µm, as for the adding of S-adenosylhomocysteine were (0.460 ± 0.005) and (27.343 ± 1.107) µm. When adding the supernatant fluid of standard strains, the biofilm thickness and the growth levels of mutant strain were much higher than those of the standards strain. The biofilm thickness and growth of both kinds of strains decreased after the addition of S-adenosylmethionine. CONCLUSIONS: luxS gene plays not only a role in quorum sensing but also a role in sulfurmetabolism.

[Application of Cre-loxP(*) system in constructing the markerless double-gene-deletion strain in Streptococcus mutans].

OBJECTIVE: To construct double gene deletions at the htrA and clpP loci on the chromosome of Streptococcus mutans (Sm) and to remove the antibiotic resistance markers with the Cre-loxP(*) site-specific recombination system. METHODS: The htrA gene was cloned into the pGEM-T-Easy TA cloning vector and then inactivated via the insertion of a kanamycin resistance cassette (lox71-Km-lox66), yielding pGEM-T-ΔhtrA/Km for deleting the htrA gene. Using the same method, the pGEM-T-ΔclpP/Sp was constructed for deleting the clpP gene. Following the transformation of pGEM-T-ΔhtrA/Km in Sm, the homologous recombination event was selected. One such mutant was transformed with a cre expression plasmid (pCrePA). The kanamycin resistance gene was then excised. The pCrePA was then easily eliminated at nonpermissive temperatures, resulting in a mutant strain (MSΔhtrA) carrying a deletion at the htrA loci without a selectable marker. This mutant was verified by PCR and DNA sequencing. Then, the clpP and spectinomycin resistance gene were deleted from MSΔhtrA, yielding markerless mutant strain lacking clpP and htrA. RESULTS: The deletion of htrA, clpP and antibiotic resistance markers were confirmed by PCR analysis and DNA sequencing. CONCLUSIONS: A mutant of Sm was constructed successfully which contained a deletion of the htrA and clpP gene without selectable marker. The Cre-loxP(*) system can be applied to Sm, which provides experimental evidence for generating markerless multiple gene deletion mutants.

[Study of vibrational spectra of humic substance in soils from the three Gorges Reservoir Area].

The six typical different parts of soils and sediments along the bank of the Three Gorges Reservoir Area (TGRA) were collected, and the humic substance isolated from the six parts of the soils and sediments' samples was separated to humic acid and fulvic acid, purified, and characterized with the combination of the Raman and IR vibrational spectroscopic technologies after cool-dried separation; through assigning the vibrational peaks in each part of the Raman and IR spectra of each sample part, the vibrational characteristics of the structures and the groups that belonged to the molecules of the humic acids and the fulvic acids in the soils and sediments of the TGRA were obtained; the changing features of the groups and structures in the humic acid and the fulvic aicd's molecules from the different soils and sediments in the TGRA were discussed with the environmental impact factors such as soil humic degree, the conditions of different soils conference, using and/or cultivating models and water level fluctuations. From the experimental results, the vibrations about C-O, C-C, and poly-hydrogen bonds dominate in the structures and the groups of each part' humic substance; the active vibration numbers in the upstream are more than in the downstream; the soil's humic degree has great effect on the formation of the humic substances' structures in soil's humic substance; the soil used as agricultural cultivating mode showed higher humic degrees in the upstream parts of the TGRA. The effect of the water level's fluctuation on the formation of the humic acid and fulvic aicd in the sediments of the TGRA is not obvious in the short time.

[Study of the fluorescence characteristics of DOM from the Yangtze River and Jialing River around Chongqing's urban areas].

The fluorescence emission and excitation emission matrix (EEM) technologies were used to characterize the dissolved organic matter (DOM) in the water body of the Yangtze River and Jialing River around the Chongqing urban areas from April to August 2008. Concerning about the accidents of the Wenchuan's Earthquake in May and Tangjiashan Yansaihu's effects in June, and the high water period time in the summer in two months of July and August, from the EEM obtained from each sampling station and time, the composition, distribution and their changing features of the DOM in the two rivers were investigated as combined with the water samples' environmental parameters such as pH, DO, DOC with EEM's fingerprint features, f(450/500) etc; finally the bio-environment behavior effects of the three types of fluorescence peaks were elaborated, where humic-like, fulvic-like, and protein-like from the five sampling stations' EEMs during the five months were given detailed representation. From the experimental results obtained, the fluorescence peaks are mainly composed of two types of fluorophores: humic-like and protein-like in the two rivers around the Chongqing urban areas during the investigation in five months, the protein-like's peaks value in Jialing River is higher than the values in the Yangtze River, and all the fluorescence peaks in the two Rivers' water body decrease more or less after the two Rivers join in Chun Tan sampling station; the protein-like peak is notably higher after the "5 x 12" earthquake period time including May and June and high water period time, which mainly originated from terrestrial sources, but its intensities decreased observably while the water bodies of the two rivers joining together in the Chao Tianmen and Chun Tan's sampling station.

[The study of dimethoate by means of vibrational and surface enhanced Raman spectroscopy on Au/Ag core-shell nanoparticles].

The vibrational structure of dimethoate, with its solid state and saturated solutions at acidic and basic conditions, was characterized with combination of means of FTIR and FT-Raman vibrational spectroscopy technology, and the comprehensive information about the dimethoate molecular groups' vibrational features was obtained. The surface enhanced Raman scattering (SERS) spectra of dimethoate at different concentrations with different acidic and basic conditions, and adsorbed on the substrate's surface of the core-shell Au/Ag nanoparticles, were also obtained. The adsorption states of dimethoate's molecule on the substrate's surface of the core-shell Au/Ag nanoparticles and the effects by the different acid-base conditions were investigated, with speculation of the adsorption mechanism. From the results, v(as)(NH), v(as)(CH3), v(O=C-N), tau(O=C-N), v(P-O), v(P=S), v(C-C) and delta(P-O-C) are the characteristic peaks of inner dimethoate structure's vibrations; and the concentration range in which dimethoate could interact with core-shell Au/Ag nanoparticles fully is about 1.0 x 10(-3) mol * L(-1) both in acidic and basic conditions. Dimethoate's molecule interacts with SERS' substrate surface mainly through P-O-C, O=C-C, (S-CH2), P=S, and CH3 structures; and the effects of dimethoate's hydrolysis path in acidic and basic conditions on the adsorption are discussed, which give some good references for the research of organophosphorus pesticides' transformations in different environmental systems.

[Study on the interaction of DNA and norfloxacin by FT-Raman].

FT-Raman and surface enhanced Raman spectra (SERS) of norfloxacin and norfloxacin capsule on the silver colloidal substrate were recorded, and the vibrational and enhanced peaks were assigned; interactions of norfloxacin and DNA were researched. The experimental results proved that the characteristic vibrations of capsule's content and NFX standard, which is v(C-F), vs(O-C-O), v(C=C), did not change, while the changes in molecular skeleton's vibrations were substantial. Additives do not affect the Raman spectroscopy of norfloxacin substantively. An analytical method for detecting norfloxacin medcine can be established by FT-Raman. Without any ions, the interaction is mainly caused by the inserting-action mode between the ring of norfloxacin and the double helix structure of DNA, and norfloxacin could interact with DNA directly, providing reliable evidence for antibacterial mechanism of quinoxalone antibiotics.

[FTIR, FT-Raman and surface enhanced Raman study of shikimic acid].

The authors reported the FT-infrared, and FT-Raman spectra of shikimic acid in solid state and in saturated liquid state in the present article. The vibrational peaks of each group in the molecular structure of shikimic acid were obtained and assigned, and furthermore, the Raman and IR vibrational shifts of the corresponding groups in the molecular structure of shikimic acid were analyzed and speculated especially. As shikimic acid mixed with silver nanoparticle the SERS-active substrate, and was adsorbed on the surface of the silver nanoparticle, we also obtained the surface enhanced Raman scattering (SERS) spectra of shikimic acid at different concentrations from 1.0 x 10(-1) to 1.0 x 10(-5) mol x L(-1), further more the adsorption state and the adsorption characteristics of shikimic acid, adsorbed on the surface of the silver nanoparticle, were studied and speculated with the SERS method and the surface selective rule together. Strong Raman signals were detected in the experiments and each group's vibrations in the molecular structure of shikimic acid were resolved with the combination of FT-infrared and FT-Raman spectroscopy. From the experiment data, the ideal optimized concentrations range is lower than 1 x 10(-3) mol x L, among which we could obtain better SERS spectra. Still, the experimental results suggested that the groups in the molecule structure of shikimic acid, such as COO-, OH-, CH2 and so on, can interact and be absorbed tightly with the surface of silver nanoparticle, and this interaction and absorption were mainly through the SERS charge transfer process mechanism of hydroxyl and carboxyl, and SERS electromagnetic process mechanism of methylene and hypomethyl together with the surface of silver nanopartical. The vibrations of alkene's C=C bond on the ring of cyclohexene in the molecule structure of shikimic showed no obvious enhancement in SERS spectra, which indicated that alkene's C=C bond on the ring of cyclohexene in the molecule structure of shikimic acid had no effective interaction and adsorption on the surface of silver nanoparticle according to the surface selective rule.

[Enriching of total flavonoids from Herba Leonuri with polyamide and macroporous resin].

OBJECTIVE: To study the enriching method of total flavonoid from Herba Leonuri with polyamide and macroporous resin. METHOD: Seven enriching and purifying methods were compared with the yield and purity as indexes. The method of enriching with polyamide and macroporous resin was confirmed and the process of purifying was determined by orthogonal design. RESULT: D101 resin is packed by wet method, the ratio of diameter to height is 1:7. After mixed with the extract liquids, the weight of wet resin increased to 3 times of the dry resin. Evaporated the wet resin to dryness, mixed well with a little of 95% ethanol and dry polyamide powder, evaporated them to dryness again. Elute with deionized water until the effluent being colourless, then loaded it on the macroporous adsorptive resin, elute with 50% ethanol, the volume of effluents was collected to 7 times of the column volume. The purity of total flavonoids reached to 23%, while the diversion rate from raw Herba Leonuri was to 69%. CONCLUSION: The process is simple and convenient, and the regeneration of resin is easy, which has a good application foreground.

[Detection of quorum-sensing pathway and construction of luxS gene allelic exchange plasmid of Streptococcus mutans].

OBJECTIVE: To detect the AI-2 quorum-sensing pathway and construct the luxS g-ene allelic exchange plasmid of Streptococcus mutans. METHODS: To detect AI-2 pathway in Streptococcus mutans, the Vibrio harveyi BB170 was used as reporter strain. The PCR fragments of the upstream and downstream regions of luxS and the Erythromycin resistance gene were amplified with the primers respectively, and these fragments were ligated into pUC19 vector with double endonuclease reaction sequentially, the ligated DNAs were transformed into Escherichia coli DH5alpha, then the reconstructed plasmids were isolated and identified by restricted endonuclease digestions. RESULTS: Streptococcus mutans Ingbritt C could induce luminescence of BB170, suggesting the presence of AI-2 quorum sensing pathway in Streptococcus mutans, and such stimulatory activity was maximal at the mid-log growth phase. The recombinant plasmid pUCluxKO was digested by PstI-BamHI, and the digest product were 1000 bp and 5000 bp. When the pUCluxKO was digested by BamHI-KpnI, the digest product were 1500 bp and 4500 bp. While it was digested by KpnI-EcoRI, the digest product were 1000 bp and 5000 bp. All PCR product was in a single belt respectively. CONCLUSIONS: The recombinant plasmid was cloned effectively and can be used in the construction of S.mutans luxS mutant.

[Study on adsorption of nonpolar R-side amino acids on silver nanoparticles by FT-SERS].

The adsorption orientation and adsorption characteristics of aliphatic-nonpolar amino acids represented by L-methionine on silver nanoparticles were studied by FT-surface-enhanced Raman spectroscopy (FT-SERS) method with different pH and concentrations. According to the experiments, in order to get the best FT-SERS effects, the optimal concentration range of the L-methionine on silver sol is between 10(-3) mol x L(-1)-10(-4)(mol x L(-1). The FT-SERS effects and adsorption model change with pH; The SERS are poor when pH is near the isoelectric point or over 11 or 3. The interaction of L-methionine with Ag nanoparticles is the physical and chemical adsorption by NH3, COO-, S, which could also be effected by pH. From the results and previous reports on this aspect, the general rules of the adsorption model and the orientation of the aliphatic-nonpolar amino acids on silver nanoparticles were speculated, which would be useful for the study of amino acids, protein, peptide, enzyme etc.