Site-specific fabrication of a melanin-like pigment through spatially confined progressive assembly on an initiator-loaded template.

Melanin-like nanomaterials have emerged in surface biofunctionalization in a material-independent manner due to their versatile adhesion arising from their catechol-rich structures. However, the unique adhesive properties of these materials ironically raise difficulties in their site-specific fabrication. Here, we report a method for site-specific fabrication and patterning of melanin-like pigments, using progressive assembly on an initiator-loaded template (PAINT), different from conventional lithographical methods. In this method, the local progressive assembly could be naturally induced on the given surface pretreated with initiators mediating oxidation of the catecholic precursor, as the intermediates generated from the precursors during the progressive assembly possess sufficient intrinsic underwater adhesion for localization without diffusion into solution. The pigment fabricated by PAINT showed efficient NIR-to-heat conversion properties, which can be useful in biomedical applications such as the disinfection of medical devices and cancer therapies.

Carbohydrate-protein interaction-based detection of pathogenic bacteria using a biodegradable self-powered biosensor.

Battery-free and biodegradable sensors can detect biological elements in remote areas. The triboelectric nanogenerator (TENG) can potentially eliminate the need for a battery by simply converting the abundant vibrations from nature or human motion into electricity. A biodegradable sensor system integrated with TENG to detect commonly found disease-causing bacteria (E. coli) in the environment is showcased herein. In this system, D-mannose functionalized 3D printed polylactic acid (PLA) with the brush-painted silver electrode was used to detect E. coli by a simple carbohydrate-protein interaction mechanism. The adsorption capacity of D-mannose is generally altered by varying the concentration of E. coli resulting in changes in resistance. Thus, the presented biosensor can detect bacterial concentrations by monitoring the output current. The PLA TENG generates an output of 70 V, 800 nA, and 22 nC, respectively. In addition, tap water and unpasteurized milk samples are tested for detecting bacteria, and the output is measured at 6 µA and 5 µA, respectively. Further, the biosensor was tested for biodegradability in soil compost by maintaining constant temperature and humidity. This study not only proposes an efficient and fast method for screening E. coli but also gives important insights into the ability to degrade and long-term reliability of TENG-based sensor platforms.

Techniques for monitoring bioavailable organic pollutants in sediment: Application of poly(methyl methacrylate) as a passive sampler.

A poly(methyl methacrylate) (PMMA) passive sampler was applied to harbor sediment to examine whether the substrate could be used as a tool to measure freely dissolved concentrations of contaminants. An ex situ method required at least 1 g of PMMA to detect freely dissolved polycyclic aromatic hydrocarbons (PAHs) in sediment with <100 ng/g dry weight. Two weeks were sufficient to reach equilibrium under 180 rpm for PAHs with a molar volume of <250 cm3/mol. For the in situ method, a deployment time of four months was sufficient to measure PAHs with a molar volume up to 250 cm3/mol in the sediment bed. The PMMA passive sampler could be used to measure the bioavailable fraction of PAHs in porewater, reflecting the complex properties of sediment with strong sorption such as black carbons.

Possibilities of poly(methyl methacrylate) as a passive sampler for determination of bioavailable concentrations in seawater.

Solvent-treated poly(methyl methacrylate) (PMMA) was recently introduced as a passive sampler for determining bioavailable concentrations, i.e., freely dissolved concentrations. However, the much knowledge required to obtain accurate bioavailable concentrations using the thus treated PMMA, applied in a marine environment, is still lacking. In this study, uptake experiments with PMMA after solvent treatment were conducted to investigate its uptake capacity and the effects of water temperature and salinity on the PMMA-water partition coefficient (KPMMA-W) for polycyclic aromatic hydrocarbons (PAHs). Thus, PMMA passive samplers preloaded with performance reference compounds were exposed to seawater to first estimate the deployment time and then to confirm if the PMMA could give the residual concentrations of PAH in mussel. The less hydrophobic PAHs (log octanol-water partition coefficient < 5.5) had higher uptake capacity of PMMA-uptake was increased by a factor of up to 10. Whereas for these PAHs the KPMMA-W values and seawater temperature showed a parabolic relationship, the effect of salinity on KPMMA-W was not observed. The less hydrophobic PAH concentrations in seawater can be measured using the PMMA passive sampler over a period of about three weeks. For the PAHs detected in both PMMA and mussel, the PAH concentrations in mussel predicted from PMMA were found to be within one order of magnitude of the measured concentrations. This, therefore, suggests that solvent-treated PMMA could be used as a passive sampler to provide information on bioavailable concentrations for less hydrophobic PAHs.

Enhanced production of oceanic dimethylsulfide resulting from CO2-induced grazing activity in a high CO2 world.

Oceanic dimethylsulfide (DMS) released to the atmosphere affects the Earth's radiation budget through the production and growth of cloud condensation nuclei over the oceans. However, it is not yet known whether this negative climate feedback mechanism will intensify or weaken in oceans characterized by high CO(2) levels and warm temperatures. To investigate the effects of two emerging environmental threats (ocean acidification and warming) on marine DMS production, we performed a perturbation experiment in a coastal environment. Two sets of CO(2) and temperature conditions (a pCO(2) of ∼900 ppmv at ambient temperature conditions, and a pCO(2) of ∼900 ppmv at a temperature ∼3 °C warmer than ambient) significantly stimulated the grazing rate and the growth rate of heterotrophic dinoflagellates (ubiquitous marine microzooplankton). The increased grazing rate resulted in considerable DMS production. Our results indicate that increased grazing-induced DMS production may occur in high CO(2) oceans in the future.

An Intelligent System Approach for Probabilistic Volume Rendering Using Hierarchical 3D Convolutional Sparse Coding.

In this paper, we propose a novel machine learning-based voxel classification method for highly-accurate volume rendering. Unlike conventional voxel classification methods that incorporate intensity-based features, the proposed method employs dictionary based features learned directly from the input data using hierarchical multi-scale 3D convolutional sparse coding, a novel extension of the state-of-the-art learning-based sparse feature representation method. The proposed approach automatically generates high-dimensional feature vectors in up to 75 dimensions, which are then fed into an intelligent system built on a random forest classifier for accurately classifying voxels from only a handful of selection scribbles made directly on the input data by the user. We apply the probabilistic transfer function to further customize and refine the rendered result. The proposed method is more intuitive to use and more robust to noise in comparison with conventional intensity-based classification methods. We evaluate the proposed method using several synthetic and real-world volume datasets, and demonstrate the methods usability through a user study.

Molecular characterization of two myoglobins of Paragonimus westermani.

Myoglobins (Mbs), globin proteins, are present in high concentrations in trematodes. In Paragonimus westermani, 2 cDNAs were found to encode Mbs. The first clone, Pwmyo1, codes a total of 149 amino acids with a calculated mass of 16.6 kDa. The second, Pwmyo2, encodes a 146-amino acid protein with a calculated mass of 16.2 kDa. The predicted secondary structures showed the presence of 8 helices, which is the basic characteristic of Mbs. Sequence alignment revealed a high homology with the other trematode Mbs. The 2 clones contained the characteristic tyrosyl residues at helical positions B10 and distal E7, which are substitutions that have been previously shown to contribute to the high oxygen affinity of Mbs. Polyclonal antibodies against the recombinant Mbs were raised with no cross-reactivity observed. Immunolocalization revealed the proteins to be distributed generally throughout the parenchymal tissues, but absent from the tegument and reproductive organs. The cell mass of the eggs of the worm stained positive to Pwmyo2 but not Pwmyo1, suggesting the stage-specific expression of these Mbs.

Molecular and biochemical characterization of hemoglobinase, a cysteine proteinase, in Paragonimus westermani.

The mammalian trematode Paragonimus westermani is a typical digenetic parasite, which can cause paragonimiasis in humans. Host tissues and blood cells are important sources of nutrients for development, growth and reproduction of P. westermani. In this study, a cDNA clone encoding a 47 kDa hemoglobinase of P. westermani was characterized by sequencing analysis, and its localization was investigated immunohistochemically. The phylogenetic tree prepared based on the hemoglobinase gene showed high homology with hemoglobinases of Fasciola hepatica and Schistosoma spp. Moreover, recombinant P. westermani hemoglobinase degradaded human hemoglobin at acidic pH (from 3.0 to 5.5) and its activity was almost completely inhibited by E-64, a cysteine proteinase inhibitor. Immunohistochemical studies showed that P. westermani hemoglobinase was localized in the epithelium of the adult worm intestine implying that the protein has a specific function. These observations suggest that hemoglobinase may act as a digestive enzyme for acquisition of nutrients from host hemoglobin. Further investigations may provide insights into hemoglobin catabolism in P. westermani.

Evaluation of taxonomic validity of four species of Acanthamoeba: A. divionensis, A. paradivionensis, A. mauritaniensis, and A. rhysodes, inferred from molecular analyses.

The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of 0.2-1.1% with each other, but more than 2% difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group II on the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.

Molecular cloning and characterization of a cDNA encoding a laminin-binding protein (AhLBP) from Acanthamoeba healyi.

Adherence of Acanthamoeba to host tissue is believed to be crucial in the establishment of amoebic keratitis or GAE. We have isolated a cDNA from a GAE-causing gymnoamoeba, Acanthamoeba healyi, encoding a protein that binds laminin by screening with a peptide G-specific DNA probe. The cDNA clone (AhLBP) was identified on the basis of sequence homology to the nonintegrin mammalian metastasis-associated 67-kDa laminin receptor (67-LR). The predicted amino acid sequence is 256 residues long with a calculated molecular mass of 28.2kDa and a theoretical pI of 5.48. Southern and Northern blot analyses suggested the gene as a single copy in A. healyi genome and expressed as a single transcript of approximately 1.0kb. Virulent strains of Acanthamoeba revealed higher level of the AhLBP mRNA expression than soil isolates. Specific binding of the purified recombinant protein to laminin was confirmed by sandwich Western blot. The polypeptide encoded by AhLBP shared substantial identity with the acidic class ribosomal proteins involved in protein synthesis. Therefore, the AhLBP may be multifunctional in A. healyi, acting as a laminin-binding molecule but also playing a role in cell division and growth. AhLBP-EGFP fusion protein expressed in A. healyi was localized mainly at the cell membrane and nucleus and at cytoplasm with lesser degree. N-terminal 64 amino acids were important for the localization at the cell membrane. This is the first description of a cDNA encoding a laminin-binding protein from protozoan parasites.