Publications

Cotton ovule in vitro cultures are a promising platform for exploring biofabrication of fibers with tailored properties. When the ovules' growth medium is supplemented with chemically synthesized cellulose precursors, it results in their integration into the developing fibers, thereby tailoring their end properties. Here, we report the feeding of synthetic glucosyl phosphate derivative, 6-deoxy-6-fluoro-glucose-1-phosphate (6F-Glc-1P) to cotton ovules growing in vitro, demonstrating the metabolic incorporation of 6F-Glc into the fibers with enhanced mechanical properties and moisture-retention capacity while emphasizing the role of molecular hierarchical architecture in defining functional characteristics and mechanical properties. This incorporation strategy bypasses the early steps of conventional metabolic pathways while broadening the range of functionalities that can be employed to customize fiber end properties. Our approach combines materials science, chemistry, and plant sciences to illustrate the innovation required to find alternative solutions for sustainable production of functional cotton fibers with enhanced and emergent properties.

Deep learning is a powerful tool for exploring large datasets and discovering new patterns. This work presents an account of a metric learning-based deep convolutional neural network (CNN) applied to an archaeological dataset. The proposed account speaks of three stages: training, testing/validating, and community detection. Several thousand artefact images, ranging from the Lower Palaeolithic period (1.4 million years ago) to the Late Islamic period (fourteenth century AD), were used to train the model (i.e., the CNN), to discern artefacts by site and period. After training, it attained a comparable accuracy to archaeologists in various periods. In order to test the model, it was called to identify new query images according to similarities with known (training) images. Validation blinding experiments showed that while archaeologists performed as well as the model within their field of expertise, they fell behind concerning other periods. Lastly, a community detection algorithm based on the confusion matrix data was used to discern affiliations across sites. A case-study on Levantine Natufian artefacts demonstrated the algorithms capacity to discern meaningful connections. As such, the model has the potential to reveal yet unknown patterns in archaeological data.

Global warming has prompted a search for new materials that capture and sink carbon dioxide (CO2). Biochar is a derivative of biomass pyrolysis and a carbon sink mainly used to improve crop production. This work explores the underlying mechanism behind biochars electric conductivity using a wide range of feedstocks and its combination with a binder (gypsum). This gypsumbiochar composite exhibits decreased density and flexural moduli with increasing biochar content, particularly after 20% w/w. Gypsumbiochar drywall-like composite prototypes display increasing shielding efficiency mostly in the microwave range as a function of biochar content, differing from other conventional metal (copper) and synthetic carbon-based materials. This narrow range of electromagnetic interference (EMI) shielding is attributed to natural alignment (isotropy) of the carbon ultrastructure (e.g., lignin) induced by heat and intrinsic interconnectivity in addition to traditional phenomena such as dissipation of surface currents and polarization in the electric field. These biomass-derived products could be used as sustainable lightweight materials in a future bio-based economy.

Production of stone artefacts using pyro-technology is known from the Middle and Upper Palaeolithic of Europe and the Levant, and the Middle Stone Age in Africa. However, determination of temperatures to which flint artefacts were exposed is impeded by the chemical and structural variability of flint. Here we combine Raman spectroscopy and machine learning to build temperature-estimation models to infer the degree of pyro-technological control effected by inhabitants of the late Lower Palaeolithic (Acheulo-Yabrudian) site of Qesem Cave, Israel. Temperature estimation shows that blades were heated at lower median temperatures (259°C) compared to flakes (413°C), whereas heat-induced structural flint damage (for example, pot-lids and microcracks) appears at 447°C. These results are consistent with a differential behaviour for selective tool production that can be viewed as part of a plethora of innovative and adaptive behaviours of Levantine hominins >300,000 years ago.

Micro/nano thermogravimetry (TG) employing MEMS has significant potential to improve the minimum sample mass and mass resolution as compared to commercial TG instruments. Although there have been a few previous reports on MEMS TG, none of them have critically analysed the obtained TG curve in detail. In this work, we have designed and fabricated a microelectromechanical thermogravimetric device (MMTG) with integrated microheaters and temperature sensors. The mass sensitivity of the device was estimated to be 0.89 pg Hz(-1) which outperforms the standard TG approaches. We tested the MMTG performance with CuSO4 center dot 5H(2)O crystals. The final mass loss ratio corresponds to the theoretically expected value, although the obtained TG curve deviated from the standard TG curve of CuSO4 center dot 5H(2)O obtained from commercial TG instruments. We attributed the deviation to the inherent temperature non-uniformity, non-isothermal conditions and temperature gradients of metallic-wire based microheaters. Finite element (FE) simulations were carried out in order to confirm and gain insights into the non-uniform heating phenomena of microheater and sample. Based on the simulation results, we propose designs that can be realized to make MEMS TG a successful enterprise.

The impressive mechanical properties of natural composites, such as nacre, arise from their multiscale hierarchical structures, which span from nano- to macroscale and lead to effective energy dissipation. While some synthetic bioinspired materials have achieved the toughness of natural nacre, current production methods are complex and typically involve toxic chemicals, extreme temperatures, and/or high pressures. Here, the exclusive use of bacteria to produce nacre-inspired layered calcium carbonate-polyglutamate composite materials that reach and exceed the toughness of natural nacre, while additionally exhibiting high extensibility and maintaining high stiffness, is introduced. The extensive diversity of bacterial metabolic abilities and the possibility of genetic engineering allows for the creation of a library of bacterially produced, cost-effective, and eco-friendly composite materials.

Multicomponent nanostructures containing purely organic or inorganic as well as hybrid organicinorganic components connected through a solid interface are, unlike conventional spherical particles, able to combine different or even incompatible properties within a single entity. They are multifunctional and resemble molecular amphiphiles, like surfactants or block copolymers, which makes them attractive for the self-assembly of complex structures, drug delivery, bioimaging, or catalysis. We have synthesized Pd@FexO heterodimer nanoparticles (NPs) to fabricate a macroporous, hydrophobic, magnetically active, three-dimensional (3D), and template-free hybrid foam capable of repeatedly separating oil contaminants from water. The Pd domains in the Pd@FexO heterodimers act as nanocatalysts for the hydrosilylation of polyhydrosiloxane and tetravinylsilane, while the FexO component confers magnetic properties to the final functional material. Pd@FexO heterodimers were synthesized by heterogeneous nucleation and growth of the iron oxide domain onto presynthesized Pd NPs at high temperatures in solution. The morphology, structure, and magnetic properties of the as-synthesized heterodimers were characterized by transmission electron microscopy (TEM), X-ray diffraction, Mössbauer spectroscopy, and a superconducting quantum interference device. The epitaxial growth of the FexO domain onto Pd was confirmed by high-resolution TEM. A potential application of the 3D hydrophobic magnetic foam was exploited by demonstrating its ability to soak oil beneath a water layer, envisioning its use in oil sampling during oil prospection drilling, or to remove oil films after oil spills.

The skeletal system of Demospongiae consists of siliceous spicules, which are composed of an axial channel containing an organic axial filament (AF) surrounded by a compact layer of hydrated amorphous silica. Here we report the ultrastructural investigations of the AF of siliceous spicules from two Demospongiae: Suberites domuncula and Tethya aurantium. Electron microscopy, electron diffraction and elemental mapping analyses on both longitudinal and transversal cross-sections yield that spicules's AF consist of a three-dimensional crystal lattice of six-fold symmetry. Its structure, which is the result of a biological growth process, is a crystalline assembly characterized by a lattice of organic cages (periodicity in the range of 6 nm) filled with enzymatically-produced silica. In general, the six-fold lattice symmetry is reflected by the morphology of the AF, which is characterized by six-fold facets. This seems to be the result of a lattice energy minimization process similar to the situation found during the growth of inorganic crystals. Our structural exploitation of three-dimensional organic lattices generated by biological systems is expected to contribute for explaining the relation between axial filament's ultrastructure and spicule's ultimate morphology. (C) 2017 Elsevier Inc. All rights reserved.

Boron's unusual properties inspired major advances in chemistry. In nature, the existence and importance of boron has been fairly explored (e.g. bacterial signaling, plant development) but its role as biological catalyst was never reported. Here, we show that boric acid [B(OH)(3)] can restore chloroperoxidase activity of Curvularia inaequalis recombinant apo-haloperoxidase's (HPO) in the presence of hydrogen peroxide and chloride ions. Molecular modeling and semi-empirical PM7 calculations support a thermodynamically highly favored (bio)catalytic mechanism similarly to vanadium haloperoxidases (V-HPO) in which [B(OH)(3)] is assumedly located in apo-HPO's active site and a monoperoxyborate [B(OH)(3)(OOH)(-)] intermediate is formed and stabilized by interaction with specific active site amino acids leading ultimately to the formation of HOCl. Thus, B(OH)(3)-HPO provides the first evidence towards the future exploitation of borons role in biological systems.

Cotton is the one of the world's most important crops. Like any other crop, cotton growth/development and fiber quality is highly dependent on environmental factors. Increasing global weather instability has been negatively impacting its economy. Cotton is a crop that exerts an intensive pressure over natural resources (land and water) and demands an overuse of pesticides. Thus, the search for alternative cotton culture methods that are pesticide-free (biocotton) and enable customized standard fiber quality should be encouraged. Here we describe a culture of Gossypium hirsutum ("Upland" Cotton) utilizing a greenhouse and hydroponics in which the fibers are morphological similar to conventional cultures and structurally fit into the classical two-phase cellulose I model with 4.19 nm crystalline domains surrounded by amorphous regions. These fibers exhibit a single crystalline form of cellulose I-I-beta, monoclinic unit cell. Fiber quality bulk analysis shows an improved length, strength, whiteness when compared with soil based cultures. Finally, we show that our fibers can be spun, used for production of non-woven fabrics and indigo-vat stained demonstrating its potential in industrial and commercial applications. (C) 2016 Elsevier Inc. All rights reserved.

Sulfite oxidase is a mitochondria-located molybdenum-containing enzyme catalyzing the oxidation of sulfite to sulfate in the amino acid and lipid metabolism. Therefore, it plays a major role in detoxification processes, where defects in the enzyme cause a severe infant disease leading to early death with no efficient or cost-effective therapy in sight. Here we report that molybdenum trioxide (MoO3) nanoparticles display an intrinsic biomimetic sulfite oxidase activity under physiological conditions, and, functionalized with a customized bifunctional ligand containing dopamine as anchor group and triphenylphosphonium ion as targeting agent, they selectively target the mitochondria while being highly dispersible in aqueous solutions. Chemically induced sulfite oxidase knockdown cells treated with MoO3 nanoparticles recovered their sulfite oxidase activity in vitro, which makes MoO3 nanoparticles a potential therapeutic for sulfite oxidase deficiency and opens new avenues for cost-effective therapies for gene-induced deficiencies.

Conventional vapor-deposition techniques for coatings require sophisticated equipment and/or high-temperature resistant substrates. Therefore bio-inspired techniques for the fabrication of inorganic coatings have been developed in recent years. Inspired by the biology behind the formation of the intricate skeletons of diatoms orchestrated by a class of cationic polyamines (silaffins) we have used surface-bound spermine, a naturally occurring polyamine, to promote the fast deposition of homogeneous, thin and transparent biomimetic SnO2 coatings on glass surfaces. The bio-enabled SnO2 film is highly photoactive, i.e. it generates superoxide radicals (O-2(-)) upon sunlight exposure resulting in a strong degradation of organic contaminants and a strong antimicrobial activity. Upon illumination the biomimetic SnO2 coating exhibits a switchable amphiphilic behavior, which - in combination with its photoactivity - creates a self-cleaning surface. The intrinsic self-cleaning properties could lead to the development of new protective, antifouling coatings on various substrates.

Owing to their physical and chemical properties, inorganic functional materials have tremendous impacts on key technologies such as energy generation and storage, information, medicine, and automotive engineering. Nature, on the other hand, provides evolution-optimized processes, which lead to multifunctional inorganicbio-organic materials with complex structures. Their formation occurs under physiological conditions, and is goverened by a combination of highly regulated biological processes and intrinsic chemical properties. Nevertheless, insights into the molecular mechanisms of biomineralization open up promising perspectives for bioinspired and biomimetic design and the development of inorganicbio-organic multifunctional hybrids. Therefore, biomimetic approaches may disclose new synthetic routes under ambient conditions by integrating the concept of gene-regulated biomineralization principles. The skeletal structures of marine sponges provide an interesting example of biosilicification via enzymatically controlled and gene-regulated silica metabolism. Spicule formation is initiated intracellularly by a fine-tuned genetic mechanism, which involves silica deposition in vesicles (silicassomes) under the control of the enzyme silicatein, which has both catalytic and templating functions. In this review, we place an emphasis on the fabrication of biologically inspired materials with silicatein as a biocatalyst.

The calcitic prisms of Pinna nobilis (Pinnidae, Linnaeus 1758) are shown to be perfect examples of a mesocrystalline material. Based on their ultrastructure and on the occurrence of an amorphous transient precursor during the early stages of prism formation, we provide evidence for the pathway of mesocrystallisation proposed by Seto et al. (2012), which proceeds not by self-organized oriented attachment of crystalline nano-bricks but by aggregation of initially amorphous nanogranules which later transform by epitaxial nucleation to a three-dimensional array of well aligned nanocrystals. We further fathom the role of a liquid amorphous calcium carbonate in biomineralisation processes and provide strong evidence for the occurrence of PILP-like intermediates during prism formation. We develop a new scenario of prism formation based on the presented findings presented findings and discuss the implications of a speculative liquid amorphous calcium carbonate (LACC) intermediate in vivo.

Ni@Fe2O3 heterodimer nanoparticles (NPs) were synthesized by thermal decomposition of organometallic reactants. After functionalization, these Ni@Fe2O3 heterodimers became water soluble. The pristine heterodimeric NPs were characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), Mossbauer spectroscopy and magnetic susceptibility measurements. A special advantage of the heterodimers lies in the fact that nanodomains of different composition can be used as catalysts for the removal of environmentally hazardous halogenated pollutants.

The metabolic energy state of sponge tissue in vivo is largely unknown. Quantitative bioluminescence-based imaging was used to analyze the ATP distribution of Suberites domuncula (Olivi 1792) tissue, in relation to differences between the cortex and the medulla. This method provides a quantitative picture of the ATP distribution closely reflecting the in vivo situation. The obtained data suggest that the highest ATP content occurs around channels in the sponge medulla. HPLC reverse-phase C-18, used for measurement of ATP content, established a value of 1.62 mu molATPg(-1) dry mass in sponge medulla, as opposed to 0.04 mu molATPg(-1) dry mass in the cortex, thus indicating a specific and defined energy distribution. These results correlate with the mitochondria localization, determined using primary antibodies against cytochrome oxidase c subunit 1 (COX1) (immunostaining), as well as with the distribution of arginine kinase (AK), essential for cellular energy metabolism (in situ hybridization with AK from S. domuncula; SDAK), in sponge sections. The highest energy consumption seemed to occur in choanocytes, the cells that drive the water through the channel system of the sponge body. Taken together, these results showed that the majority of energetic metabolism in S. domuncula occurs in the medulla, in the proximity of aqueous channels.

We describe the reconstitution of apo-horse radish peroxidase (apo-HRP) onto TiO2 nanorods functionalized with a multifunctional polymer. After functionalization, the horse radish peroxidase (HRP) functionalized TiO2 nanorods were well dispersible in aqueous solution, catalytically active and biocompatible, and they could be used to quantify and image H2O2 which is a harmful secondary product of cellular metabolism. The shape, size and structure of TiO2 nanorods (anatase) were analyzed by transmission electron microscopy (TEM), high resolution TEM (HRTEM), electron diffraction (ED) and X-ray diffraction (XRD). The surface functionalization, HRP reconstitution and catalytic activity were confirmed by UV-Vis, FT-IR, CLSM and atomic force microscopy (AFM). Biocompatibility and cellular internalization of active HRP reconstituted TiO2 nanorods were confirmed by a classical MTT cytotoxicity assay and confocal laser scanning microscopy (CLSM) imaging, respectively. The intracellular localization allowed H2O2 detection, imaging and quantification in HeLa cells. The polymer functionalized hybrid system creates a complete sensor including a "cell positioning system'' in each single particle. The flexible synthetic concept with functionalization by post-polymerization modification allows introduction of various dyes for sensitisation at different wavelengths and introduction of various anchor groups for anchoring on different particles.

The gold standard for bone reconstruction is the use of autogeneic grafts from various donor regions, since they possess osteoinductive as well as osteoconductive potential. Only a few synthetic materials possess/display properties that allow optimal bone reconstitution. Previously, we showed that the natural product, bio-silica, comprises osteoinductive, and probably also osteoconductive activity. Bio-silica is formed enzymatically via silicatein; this enzyme has been isolated from siliceous sponges and has also been cloned and prepared recombinantly. In the present study, silicatein was encapsulated together with its substrate, sodium metasilicate, in poly(D, L-lactide)/poly(vinyl pyrrolidone)-based microspheres, termed silicatein-and-silica-containing microspheres (SSMs). The deposition of silica and the kinetics of silicatein release from the microspheres are given. Furthermore, SSMs were successfully embedded in a poly(vinyl pyrrolidone)/starch-based matrix, termed plastic-like filler matrix containing silicic acid (PMSA). A blend of SSM and PMSA forms a biocompatible, moldable, and biodegradable functional implant material that hardens at a controlled and clinically suitable rate within approximately 30 min to 6 h to implants that were tightly integrated in artificial defects of rabbit femurs. Until now no toxic reactions caused by the silicatein have been observed in vitro or in vivo. We assume that the data given here contribute to a successful introduction of the silicatein/bio-silica-based implant materials to the field of regenerative medicine.

Primmorphs (a three-dimensional sponge primary cell culture system) have been revealed to be a cell/tissue nano-factory for the production of tailor-made hybrid nanostructures. Growth of primmorphs is stimulated by the presence of a titanium alkoxide precursor tolerating titania (TiO2) concentrations up to 250 mu M. The presence and activity of silicatein in primmorphs has been analyzed by gel electrophoresis and Western blotting. Results of studies by scanning transmission electron microscopy with energy-dispersive X-ray spectroscopy have revealed silica and titania to be co-localized on nanosized spicules. Our findings suggest that the incorporation of titania into the nanosized spicule is enzymatically mediated via active silicatein in an orchestrated mechanism.

Previously, the cDNA and the respective gene for a presumed tauropine dehydrogenase (TaDH) from Suberites domuncula (GenBank accession nos. AM712888, AM712889) had been annotated. The conclusion that the sequences encode a TaDH had been inferred from the 68% identity with the TaDH protein from the marine demosponge Halichondria japonica. However, subsequent enzymatic assays shown here indicate that the presumed S. domuncula opine dehydrogenase is in fact a strombine dehydrogenase (StDH). The enzyme StDH is highly specific for glycine and is inhibited by an excess of the substrate pyruvate. Besides kinetic data, we report in this study also on the predicted tertiary and quaternary structure of the sponge StDH. It is concluded that the dimer (75 kDa) has a novel structure, distinguishing it from other known marine invertebrate OpDHs that exist as monomers. (C) 2009 Elsevier Inc. All rights reserved.

High-resolution microscopy shows that during the initial stages of demosponge spicule formation, a primordial crystalline structure is formed within the axial filament. The recently developed electron diffraction tomography technique reveals that the nanorods have a layered structure that matches smectitic phyllosilicates. These intracellular nanorods have been considered as precursors of mature spicules.

A chemically specific and facile method for the biofunctionalization of WS2 nanotubes (NT-WS2) is reported. The covalent modification strategy is based on the affinity of the nitrilotriacetic acid (NTA) side chain, which serves as a ligand for the surface binding to NT-WS2 and simultaneously as an anchor group for the binding of His-tagged proteins to the polymer backbone. The polymer functionalized WS2 nanotubes can be solubilized either in water or organic solvents; they are stable for at least one week. The probes were characterized by FT-IR and UV-vis spectroscopy. The immobilization of silicatein, a hydrolytic protein encountered in marine sponges, was visualized by scanning force microscopy (SFM) and confocal laser scanning microscopy (CLSM). The formation of the biotitania coating mediated by the immobilized silicatein onto the surface was characterized by scanning electron microscopy (SEM), and transmission electron microscopy (TEM).

We describe the self-assembly of discrete SiO(2) nanofibers via grafting of silicatein side chains to a polymer backbone. The covalent binding of silicatein to the backbone of the polymer is based on the affinity of the nitrilotriacetic acid (NTA) side chain, which serves as a ligand for the immobilization of His-tagged silicatein. The surface charge and the bulkiness of the protein moieties prevent the entropically favoured coil formation of the polymer and force it to adopt an open chain structure after hydrolysis of the silica precursors. The probes were characterized by scanning force microscopy (SFM) and optical light microscopy. Surface complexation of the resulting silica nanoparticles by the polymer-bound silicatein were analysed using high resolution tranmission electron microscopy (HRTEM).

Silicatein, a hydrolytic protein encountered in marine sponges, was immobilized on maghemite (gamma-Fe2O3) nanoparticles that were surface functionalized with a reactive mulfunctional polymer. This polymer carries an anchor group based on dopamine which is capable of binding to the gamma-Fe2O3 Surface and a reactive functional group which allows binding of various biomolecules onto inorganic nanoparticles. This functional nitrilotriacetic acid (NTA) group allows immobilization of His-tagged silicatein on the surface of the gamma-Fe2O3 nanoparticles. The surface-bound protein retains its native hydrolytic activity to catalyze formation of silica through copolymerization of alkoxysilanes Si(OR)(4). Functionalization of the magnetic nanoparticles and the architecture of the SiO2-coated gamma-Fe2O3 nanoparticles was confirmed by TEM studies as well as by FT-IR and optical microscopy.

An appreciation of the potential applications of molecular biology is of growing importance in many areas of life sciences, including marine biology. During the past two decades, the development of sophisticated molecular technologies and instruments for biomedical research has resulted in significant advances in the biological sciences. However, the value of molecular techniques for addressing problems in marine biology has only recently begun to be cherished. It has been proven that the exploitation of molecular biological techniques will allow difficult research questions about marine organisms and ocean processes to be addressed. Marine molecular biology is a discipline, which strives to define and solve the problems regarding the sustainable exploration of marine life for human health and welfare, through the cooperation between scientists working in marine biology, molecular biology, microbiology and chemistry disciplines. Several success stories of the applications of molecular techniques in the field of marine biology are guiding further research in this area. In this review different molecular techniques are discussed, which have application in marine microbiology, marine invertebrate biology, marine ecology, marine natural products, material sciences, fisheries, conservation and bio-invasion etc. In summary, if marine biologists and molecular biologists continue to work towards strong partnership during the next decade and recognize intellectual and technological advantages and benefits of such partnership, an exciting new frontier of marine molecular biology will emerge in the future. (C) 2008 Elsevier Inc. All rights reserved.

Double stranded RNA polyinosinic-polycytidlylic acid (poly(IC)) immobilized onto y-Fe₂O₃ nanoparticles with the use of a multifunctional polymer linker is analyzed. Among the most potent dsRNAs is the synthetic dsRNA poly(IC), which consists of a pair of strands of polyinosinic and polycytidylic acids. Poly(IC) shows antitumor and anti- viral activity and recently entered into phase II clinical trials for patients with malignant gliomas. The immobilization of biomolecules such as poly(IC) onto insoluble supports or nanoparticles is an important tool for the fabrication of functional materials or devices. Ferrimagnetic y-Fe ₂O₃ nanoparticles were synthesized by co-precipitation of ferrous and ferric ions in sodium hydroxide solution. The efficiency and stability of binding between the amine-functionalized iron oxide nanoparticles and poly(IC) was studied by electrophoresis.