Uncyclized xanthommatin is a key ommochrome intermediate in invertebrate coloration.

Ommochromes are widespread pigments that mediate multiple functions in invertebrates. The two main families of ommochromes are ommatins and ommins, which both originate from the kynurenine pathway but differ in their backbone, thereby in their coloration and function. Despite its broad significance, how the structural diversity of ommochromes arises in vivo has remained an open question since their first description. In this study, we combined organic synthesis, analytical chemistry and organelle purification to address this issue. From a set of synthesized ommatins, we derived a fragmentation pattern that helped elucidating the structure of new ommochromes. We identified uncyclized xanthommatin as the elusive biological intermediate that links the kynurenine pathway to the ommatin pathway within ommochromasomes, the ommochrome-producing organelles. Due to its unique structure, we propose that uncyclized xanthommatin functions as a key branching metabolite in the biosynthesis and structural diversification of ommatins and ommins, from insects to cephalopods.

Nano silver chloride and alginate incorporated composite copolymer adsorbent for adsorption of a synthetic dye from water in a fixed bed column and its photocatalytic reduction.

Several functional adsorbents were synthesized by copolymerizing acrylic acid (AA) and hydroxyethyl methacrylate (HEMA) in the presence of sodium alginate in water. Silver chloride nano particles (AgClNP) were also incorporated in the polymer matrix by coprecipitation method. The resulting composite type functional adsorbents were used for adsorption of a synthetic dye, namely, brilliant cresyl blue (BCB) from water in batch and in continuous mode in a fixed bed column. The formation of the adsorbent, AgClNP and its incorporation in the polymer matrix were characterized while pH sensitivity was studied with pH reversibility test and point zero charge (PZC) study. The network parameters were also determined. The synthesis and process parameters were optimized with a central composite design (CCD) of response surface methodology (RSM) with respect to adsorption capacity as response in a batch study and the adsorbent showing optimum performance was further used for column study in a fixed bed. The effect of bed height, influent dye concentration and volumetric flow rate on dye adsorption were studied and the adsorption data showed close fitting to Adam-Bohart and Clark model. The photocatalytic degradation of the heterocyclic dye by the AgClNP incorporated adsorbent in the presence of UV radiation was also studied.

Questiomycins, Algicidal Compounds Produced by the Marine Bacterium Alteromonas sp. D and Their Production Cue.

Questiomycin A (1) along with three new compounds, questiomycins C-E (2-4), were isolated from culture of Alteromonas sp. D, an algicidal marine bacterium, guided by algal lethality assay using the raphidophyte, Chattonella antiqua, one of the causative organisms of harmful algal bloom. The structures of 1-4 were assigned on the basis of their spectrometric and spectroscopic data. Compounds 1 to 4 exhibited algicidal activity against C. antiqua with LC50 values ranging from 0.18 to 6.37 M. Co-cultivation experiment revealed that 1 was produced only when the microalgae and the bacterium are in close contact, suggesting that some interactions between them trigger the biosynthesis of questiomycins. These results suggested that the algicidal bacteria such as Alteromonas sp. D can control microalgae chemically in marine ecosystem.

Residue analysis of tebufenozide and indoxacarb in chicken muscle, milk, egg and aquatic animal products using liquid chromatography-tandem mass spectrometry.

We developed an analytical method using liquid-liquid extraction (LLE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to detect and quantify tebufenozide (TEB) and indoxacarb (IND) residues in animal and aquatic products (chicken muscle, milk, egg, eel, flatfish, and shrimp). The target compounds were extracted using 1% acetic acid (0.1% acetic acid for egg only) in acetonitrile and purified using n-hexane. The analytes were separated on a Gemini-NX C18 column using (a) distilled water with 0.1% formic acid and 5 mm ammonium acetate and (b) methanol with 0.1% formic acid as the mobile phase. All six-point matrix-matched calibration curves showed good linearity with coefficients of determination (R2 ) ≥0.9864 over a concentration range of 5-50 µg/kg. Intra- and inter-day accuracy was expressed as the recovery rate at three spiking levels and ranged between 73.22 and 114.93% in all matrices, with a relative standard deviation (RSD, corresponding to precision) ≤13.87%. The limits of quantification (LOQ) of all target analytes ranged from 2 to 20 µg/kg, which were substantially lower than the maximum residue limits (MRLs) specified by the regulatory agencies of different countries. All samples were collected from different markets in Seoul, Republic of Korea, and tested negative for tebufenozide and indoxacarb residues. These results show that the method developed is robust and may be a promising tool to detect trace levels of the target analytes in animal products.

New phenoxazinone-related alkaloids from strain Streptomyces sp. KIB-H1318.

Chemical investigation of a strain Streptomyces sp. KIB-H1318 isolated from soil sample led to the discovery of three new phenoxazinone-related alkaloids 1-3, as well as two known analogs exfoliazone (4) and viridobrunnine A (5). Their structures were determined on the basis of extensive spectroscopic analysis. The antimicrobial activity and cytotoxicity of the isolates were assayed. Exfoliazone and viridobrunnine A exhibited minor antibacterial activity against Escherichia coli ATCC 8099, Bacillus subtilis ATCC 6633, and Staphylococcus aureus ATCC 6538. Compound 2 exhibited low cytotoxicity against two human cancer cell lines HeLa and SW480 with the IC50 values of 36.8 and 37.8 µM, respectively.

Industrial prune processing and its effect on pesticide residue concentrations.

The aim of this study was to determine the insecticide residue processing factor (PF) from plums to prunes and the effect of the industrial processing of prunes residue concentrations. Our results show an increase of insecticide concentrations during plum dehydration that is explained by fruit water loss; however, the normalized insecticide residue concentration, based on plum dry weights to compensate dehydration, was reduced. The water washing and tenderizing of prunes produced insecticide residue reductions of 22.9 ±â€¯4.5% and 21.9 ±â€¯4.2%, respectively. PF were: 1.157, 1.872, 1.316, 0.192, 2.198, 0.775 and 0.156 for buprofezin, l-cyhalothrin, spirodiclofen, indoxacarb, acetamiprid, imidacloprid and emamectin benzoate, respectively, being directly related to water solubility, aqueous hydrolysis and degradation point and inversely related to molecular mass and melting point. In plums for the dehydrated agroindustry the final product is prunes, therefore, it is crucial to consider the PF to determine the specific preharvest interval for this important agroindustry.

Removal of Two Neonicotinoid Insecticides and Mineralization of 14C-Imidacloprid in Biomixtures.

Environmental contamination with neonicotinoid insecticides represents an issue of wide concern due to their negative effects on pollinators. The goal of this work was to evaluate the potential use of biomixtures employed in biopurification systems (BPS) to remove two neonicotinoid pesticides, imidacloprid and thiamethoxam, from wastewater of agricultural origin. The removal was assayed by quantification of the parent compounds and the detection of putative transformation products of imidacloprid by means of LC-MS/MS, and mineralization of radiolabeled imidacloprid. Two biomixtures (B1, B2) were prepared using coconut fiber, compost and two soils pre-exposed to imidacloprid (volumetric composition 50:25:25). After spiking of neonicotinoids and 228 days of treatment, the removal ranged from 22.3%-30.3% and 38.6%-43.7% for imidacloprid and thiamethoxam, respectively. Transformation products imidacloprid-urea, desnitro-imidacloprid and desnitro-olefin-imidacloprid were detected in both biomixtures. The mineralization of 14C-imidacloprid revealed DT50 (mineralization half-lives) values of 3466 and 7702 days in the biomixtures B1 and B2, respectively, markedly lower than those in the soil used in their preparation (8667 and 9902 days, respectively). As demonstrated by these findings, the high persistence of these compounds in the BPS suggests that additional biological (or physicochemical) approaches should be explored in order to decrease the impact of neonicotinoid-containing wastewater of agricultural origin.

Oxygen-dependence of mitochondrial ROS production as detected by Amplex Red assay.

The initial rates of superoxide plus hydrogen peroxide (ROS) generation by intact or permeabilized rat heart mitochondria and coupled inside-out bovine heart submitochondrial particles (SMP) oxidizing NAD-dependent substrates, NADH, and succinate were measured by detecting resorufin formation in the Amplex Red assay at various oxygen concentrations. Linear dependences of the initial rates on oxygen concentration within the range of ~125-750 µM were found for all significant mitochondrial generators, i.e. the respiratory complexes and ammonium-stimulated dihydrolipoamide dehydrogenase. At lower oxygen concentrations upon its decrease from air saturation level to zero, the time-course of resorufin formation by SMP catalyzing coupled oxidation of succinate (the total ROS production by respiratory complexes II and III and by the reverse electron transfer (RET)-mediated by complex I) also corresponds to the linear dependence on oxygen with the same first-order rate constant determined in the initial rate studies. Prolonged incubation of SMP generating succinate-supported complex I-mediated ROS affected neither their NADH oxidase nor ROS generating activity. In contrast to SMP significant deviation from the first-order oxygen dependence in the time-course kinetics during coupled oxidation of succinate by intact mitochondria was evident. Complex I catalyzes the NADH:resorufin oxidoreductase reaction resulting in formation of colorless reduced resorufin. Hydrogen peroxide oxidizes reduced resorufin in the presence of peroxidase, thus showing its dihydroresorufin peroxidase activity. Combined NADH:resorufin reductase and dihydroresorufin peroxidase activities result in underestimation of the amount of hydrogen peroxide generated by mitochondria. We conclude that only initial rates of the mitochondrial ROS production, not the amount of resorufin accumulated, should be taken as the reliable measure of the mitochondrial ROS-generating activity, because of the cycling of the oxidized and reduced resorufin during Amplex Red assays fed by NADH and other possible reductant(s) present in mitochondria.

Application of magnetic molecularly imprinted polymers for extraction of imidacloprid from eggplant and honey.

A magnetic molecularly imprinted polymer (MMIP) adsorbent for imidacloprid was prepared using non-covalent approach with functionalized nano Fe3O4 particles (magnetic cores), imidacloprid (template), acrylic acid (functional monomer), ethylene glycol dimethacrylate (cross linker) and azobisisobutyronitrile (initiator) and used for selective separation of imidacloprid from honey and vegetable samples. The polymers were characterized using FT-IR spectroscopy, SEM and TEM images. For analysis of imidacloprid LC-MS/MS equipment was used. Adsorption kinetics was best explained by pseudo-second-order kinetic model. Adsorption data fitted well into linearized Freundlich equation (R2 > 0.98). Scatchard plot analysis indicates the presence of two classes of binding sites in the MMIPs with the Cmax of 1889.6 µg g-1 and 65448.9 µg g-1, respectively. MMIPs demonstrated much higher affinity for imidacloprid over structurally similar analogues acetamiprid (α = 23.59) and thiamethoxam (α = 17.15). About 87.1 ±â€¯5.0% and 90.6 ±â€¯5.6% of the added imidacloprid was recovered from MMIPs in case of fortified eggplant and honey samples, respectively.

Thin layer chitosan-coated cellulose filter paper as substrate for immobilization of catalytic cobalt nanoparticles.

A facile approach utilizing synthesis of cobalt nanoparticles in green polymers of chitosan (CS) coating layer on high surface area cellulose microfibers of filter paper (CFP) is described for the catalytic reduction of nitrophenol and an organic dye using NaBH4. Simple steps of CFP coating with 1wt% CS aqueous solution followed by Co2+ ions adsorption from 0.2M CoCl2 aqueous solution were carried out to prepare pre-catalytic strips. The Co2+ loaded pre-catalytic strips of CS-CFP were treated with 0.19M NaBH4 aqueous solution to convert the ions into nanoparticles. Successful Co nanoparticles formation was assessed by various characterization techniques of FESEM, EDX and XRD analyzes. TGA analyses were carried out on CFP, CS-CFP, and Co-CS-CFP for the determination of the amount of Co particles formed on the CS-FP, and to track their thermal properties. Furthermore, we demonstrated that the Co-CS-CFP showed an excellent catalytic activity and reusability in the reduction reactions a nitroaromatic compound of 2,6-dintirophenol (2,6-DNP) and brilliant cresyl blue (BCB) dye by NaBH4. The Co-CS-CFP catalyzed the reduction reactions of 2,6-DNP and BCB by NaBH4 with psuedo-first order rate constants of 0.0451 and 0.1987min-1, respectively.

Successive pH- and heat-induced homogenous liquid-liquid extraction.

A simple and efficient analytical method known as pH- and heat-induced homogenous liquid-liquid extraction combined with high-performance liquid chromatography has been successfully developed for the extraction and determination of neonicotinoid pesticides in aqueous samples. In this method, a few mL of a water-miscible basic extraction solvent is mixed with a high volume of an aqueous phase containing the analytes and passed through a tube which a portion of the tube is filled with sodium carbonate as a separating agent. By passing the solution, salt is dissolved and the fine droplets of the extraction solvent are formed. The produced droplets go up through the remained solution and collect as a separated layer. In the following, the collected organic phase is removed and placed into a micro tube. Then it is heated in a water bath to form two phases. Several experimental parameters that influence extraction efficiency such as type and volume of extraction solvent, type of phase separation agent, temperature, and extraction time were investigated. Under the optimum conditions, the extraction recoveries and enrichment factors ranged between 51 and 81% and 680 and 1080, respectively. Calibration curves showed a high-level of linearity for all target analytes with coefficients of determination ranging between 0.997 and 0.999. The repeatability of the proposed method expressed as relative standard deviation varied between 3 and 5% (n=6, C=50µgL(-1)), and the detection limits were in the range of 0.52-1.0µgL(-1). Finally, the performance of the method was evaluated by analyzing the selected pesticides in different fruit juice and vegetable samples.

Contributions of Phenoxazone-Based Pigments to the Structure and Function of Nanostructured Granules in Squid Chromatophores.

Understanding the structure-function relationships of pigment-based nanostructures can provide insight into the molecular mechanisms behind biological signaling, camouflage, or communication experienced in many species. In squid Doryteuthis pealeii, combinations of phenoxazone-based pigments are identified as the source of visible color within the nanostructured granules that populate dermal chromatophore organs. In the absence of the pigments, granules experience a reduction in diameter with the loss of visible color, suggesting important structural and functional features. Energy gaps are estimated from electronic absorption spectra, revealing highest occupied molecular orbital (HOMO)-lowest unoccupied molecular orbital (LUMO) energies that are dependent upon the varying carboxylated states of the pigment. These results implicate a hierarchical mechanism for the bulk coloration in cephalopods originating from the molecular components confined within in the nanostructured granules of chromatophore organs.

Highly sensitive wavelength-dependent nonaqueous capillary electrophoresis for simultaneous screening of various synthetic organic dyes.

A novel multi-wavelength nonaqueous capillary electrophoresis (MW-NACE) technique based on wavelength-dependent laser-induced fluorescence (LIF) detection was investigated for the simultaneous screening of various synthetic organic dyes. Multi-wavelength excitation light sources were utilized to excite different organic dyes [e.g., 543 nm for crystal violet (CV), methyl violet B (MVB), methyl violet B base (MBB), rhodamine 6G (R6G), and rhodamine B base (RBB); 635 nm for nile blue A (NBA) and methylene blue (MB)] simultaneously. Using a nonaqueous buffer system composed of 15 mM sodium borate and 835 mM acetic acid in 100% ethanol (pH=5.4), all dyes were analyzed within 15 min with excellent resolution (R≥4.0) under an electric field of 500 V/cm. Calibration curves showed excellent linearity with square of correlation coefficients (r(2)) greater than 0.9908 over wide dynamic ranges of 0.4-50 µM for CV, 0.8-50 µM for MVB, 1.5-50 µM for MBB, 0.08-5 nM for R6G, 0.06-10 µM for MB, 0.02-10 µM for NBA, and 0.13-10 pM for RBB. The detection limits (S/N=3) of 40 fM to 0.5 µM were 10-200,000 times lower than those of previous detection methods. While adjacent peaks were not well distinguished with baseline separation in a single capillary, the devised technique was faster and more sensitive than conventional aqueous and nonaqueous CE approaches, thereby enabling the quantitative analysis of various dyes based on wavelength-dependent fluorescence detection with different excitation wavelengths.

Thermoresponsive cellulose ether and its flocculation behavior for organic dye removal.

A thermoresponsive polymer, 2-hydroxy-3-butoxypropyl hydroxyethyl cellulose (HBPEC), was prepared by grafting butyl glycidyl ether (BGE) onto hydroxyethyl cellulose (HEC). The lower critical solution temperature (LCST) and critical flocculation temperature (CFT) of HBPEC were varied by changing the molar substitution (MS) and salt concentrations. Transmission electron microscopy (TEM) images and fluorescence spectroscopy showed that HBPEC can assemble into micelles. Additionally, using Nile Red as a model dye, the performance of HBPEC for the removing Nile Red from aqueous solutions via cloud point extraction procedures was investigated in detail. The encapsulation behavior of dye in the aqueous solution of HBPEC was studied by fluorescence spectroscopy and fluorescence microscope. The experimental results indicated that 99.4% of dye was removed from the aqueous solutions, and the HBPEC was recycled and reused easily, Furthermore, the recycle efficiency (RE) and maximum loading capacity portrayed little loss with the number of cycles.

[Determination of chlorfenapyr and indoxacarb in tea by gas chromatography-mass spectrometry with solid phase extraction].

OBJECTIVE: To establish a method for simultaneous determination of chlorfenapyr and indoxacarb in tea by gas chromatography-mass spectrometry. METHODS: The tea samples were homogenized and extracted with acetonitrile. Extracts obtained through centrifugation were cleaned up by CARB / NH2 cartridges, and further purified with SLH cartridges. After separated by DB-5MS capillary column( 30 m × 0. 25 mm × 0. 25µm), the analytes were measured by gas chromatography-mass spectrometry in selective ion monitoring( SIM) mode and quantified by external standard method. RESULTS: The linear range was 0. 10- 10 µg / m L for both of the two pesticides. The detection limits of chlorfenapyr and indoxacarb in tea samples were 0. 01 and 0. 008 mg / kg, and the quantitation limits were 0. 03 and 0. 025 mg / kg, respectively. The recoveries were from75. 6% to 92. 7%, and the relative standarddeviations( RSDs) were 3. 6%- 11. 4%( n =6). CONCLUSION: The proposed method has good purification effect and high accuracy, which is capable for simultaneously detecting the chlorfenapyr and indoxacarb in tea samples.

Adsorption of cinnabarinic acid from culture fluid with magnetic microbeads.

In this study, antimicrobial pigment cinnabarinic acid (CA) was produced from Pycnoporus cinnabarinus in laboratory-scale batch cultures. Magnetic poly(ethylene glycol dimethacrylate-N-methacryloyl-l-tryptophan methyl ester) [m-poly(EGDMA-MATrp)] beads (average diameter = 53-103 µm) were synthesized by copolymerizing of N-methacryloyl-l-tryptophan methyl ester (MATrp) with ethylene glycol dimethacrylate (EGDMA) in the presence of magnetite (Fe3O4) and used for the adsorption of CA. The m-poly(EGDMA-MATrp) beads were characterized by N2 adsorption/desorption isotherms (Brunauer Emmet Teller), X-ray photoelecron spectroscopy, scanning electron microscopy, infrared spectroscopy, thermal gravimetric analysis, electron spin resonance and swelling studies. The efficiency of m-poly(EGDMA-MATrp) beads for separation of CA from culture fluid was evaluated. The effects of pH, initial concentration, contact time and temperature on adsorption were analyzed. The maximum CA adsorption capacity of the m-poly(EGDMA-MATrp) beads was 272.9 mg g(-1) at pH 7.0, 25 °C. All the isotherm data can be fitted with the Langmuir, Freundlich and Dubinin-Radushkevich isotherm models. The adsorption process obeyed pseudo-second-order kinetic model. Thermodynamic parameters ΔH = 5.056 kJ mol(-1), ΔS = 52.44 J K(-1) mol(-1) and ΔG = -9.424 kJ mol-(1) to -11.27 kJ mol-(1) with the rise in temperature from 4 to 40 °C indicated that the adsorption process was endothermic and spontaneous.

1,3-Oxazin-6-one Derivatives and Bohemamine-Type Pyrrolizidine Alkaloids from a Marine-Derived Streptomyces spinoverrucosus.

Two new 1,3-oxazin-6-one derivatives (1 and 2) and six new bohemamine-type pyrrolizidine alkaloids (3-8) were isolated from the marine-derived Streptomyces spinoverrucosus strain SNB-048. Their structures including the absolute configurations were fully elucidated on the basis of spectroscopic analysis, ECD spectra, quantum chemical calculations, and chemical methods. Compounds 1 and 2 possess a γ-lactam moiety and a 1,3-oxazin-6-one system.

Salinazinones A and B: Pyrrolidinyl-Oxazinones from Solar Saltern-Derived Streptomyces sp. KMF-004.

Salinazinones A (1) and B (2), two unprecedented pyrrolidinyl-oxazinones, were isolated from the culture broth of Streptomyces sp. KMF-004 from a solar saltern at Aphae Island, Korea. The structures of these salinazinones, which are unusual and consist of 2-methylpropenyl-1,3-oxazin-6-one bearing 1-oxopyrrolidinyl substituents, were assigned by spectral and chemical analyses using Mosher's method, circular dichroism (CD), and calculated ECD. Salinazinones are the first examples of a natural alkaloid class composed of an oxazinone-pyrrolidone conjugate.

Synthesis, characterization, swelling and dye adsorption properties of starch incorporated acrylic gels.

Several hydrogels were prepared by a free radical polymerization of acrylic acid (AA), sodium acrylate (SA) and AA/hydroxy ethyl methacrylate (HEMA) in the presence of starch in water. These starch incorporated acrylic gels were prepared by varying the concentration of the initiator, monomer, crosslinker and the starch. The resulting gels were characterized by FTIR, SEM, XRD, DTA-TGA, pH at point zero charge (PZC), swelling and the diffusion in water. The gels showed high adsorption and removal% of Safranine T (ST) and Brilliant Cresyl Blue (BCB) dyes from water. The swelling and the adsorption data were fitted to different kinetic models and isotherms. Amongst the three kinds of gels, the starch incorporated sodium polyacrylate gel showed the highest adsorption of 9.7-85.3mg/L (97-61% removal) of BCB dye and 9.1-83mg/L (91-60% removal) of ST dye for a feed dye concentration of 10-140mg/L.