Peptide engineered microcantilevers for selective chemical force microscopy and monitoring of nanoparticle capture.

Engineered peptides capable of binding to silica have been used to provide contrast in chemical force microscopy and tested for their capacity to selectively capture silica nanoparticles (NPs). Gold coated atomic force microscopy (AFM) microcantilevers with integrated tips and colloidal probes were functionalized with engineered peptides through a thiol group of a terminal cysteine which was linked via a glycine trimer to a 12-mer binding sequence. The functionalized probes demonstrated a significantly increased binding force on silicon oxide areas of a gold-patterned silicon wafer, whereas plain gold probes, and those functionalized with a random permutation of the silica binding peptide motif or an all-histidine sequence displayed similar adhesion forces to gold and silicon oxide. As the functionalized probes also allowed contact mode imaging subsequently to the adhesion mapping, also the associated friction contrast was measured and found to be similar to the adhesion contrast. Furthermore, the adsorption of silica NPs onto planar gold surfaces functionalized in the same manner was observed to be selective. Notably, the surface coverage with silica NPs was found to decrease with increasing pH, implying the importance of electrostatic interactions between the peptide and the NPs. Finally, the adsorption of silica NPs was monitored via the decrease in fundamental resonance frequency of an AFM microcantilever functionalized with silica binding peptides.

Evaluations of the Carcinogenicity of Carbon Nanotubes, Fluoro-Edinite, and Silicon Carbide by the International Agency for Research on Cancer (IARC).

We reported the evaluations of the carcinogenicity of fluoro-edinite, silicon carbide, and carbon nanotubes performed by IARC working group in October 2014. For carbon nanotubes (CNTs), multi-walled carbon nanotube (MWCNT)-7 was classified as Group 2B, and MWCNTs without MWCNT-7 and single-walled carbon nanotubes (SWCNTs) were classified as not classifiable in terms of their carcinogenicity to humans. There is sufficient evidence of carcinogenicity for MWCNT-7 in experimental animals, limited evidence for other MWCNTs, and inadequate evidence for SWCNTs. The mechanic evidence for CNTs was not strong. Fluoro-edinite was classified as carcinogenic to humans (Group 1) on the basis of sufficient evidence of carcinogenicity to humans and experimental animals. Silicon carbide was classified into silicon carbide fibers and whiskers. Silicon carbide fibers were evaluated as possibly carcinogenic to humans (Group 2B) on the basis of limited evidence of carcinogenicity to humans. Silicon carbide whiskers were evaluated as probably carcinogenic to humans (Group 2A) on the basis of sufficient evidence of carcinogenicity to experimental animals and the similarity of their physicochemical properties to those of asbestos in terms of the mechanism of carcinogenicity. We report the process of progression in meeting and discuss how to determine the evidence and the evaluation of the carcinogenicity of the three materials.

An evaluation of zinc bearing palygorskite inclusion on the growth performance, mineral content, meat quality, and antioxidant status of broilers.

The current study was conducted to investigate the effect of zinc (Zn) bearing palygorskite (ZnPal) inclusion on the growth performance, mineral content, meat quality, and antioxidant status of broilers. A total of 240 one-day-old Arbor Acres broiler chicks were randomly allocated into 5 dietary treatments with 6 replicates of 8 chicks. Broilers in the 5 treatments were fed a basal diet supplemented with 0, 20, 40, 60, and 80 mg/kg Zn diet in the form of ZnPal for 42 d, respectively. Birds exhibited similar average daily gain (ADG), average daily feed intake (ADFI), and feed/gain ratio (F:G) among groups during the 42-day study (P>0.05). ZnPal supplementation linearly increased iron (Fe) (P=0.031) and magnesium (Mg) (P=0.002) content in the pectoralis major muscle. Similarly, the inclusion of ZnPal tended to increase Zn content in the thigh (P=0.072) and linearly increase Zn content in the pectoralis major muscle (P=0.055). The concentration of copper (Cu) in the thigh was linearly decreased by ZnPal inclusion (P=0.011). Meanwhile, a quadratic trend for reduced Cu content was observed in the pectoralis major muscle (P=0.074) and thigh (P=0.082), respectively. The supplementation of ZnPal linearly reduced cooking loss in the pectoralis major muscle (P=0.013), and linearly (P=0.029) and quadratically (P=0.034) decreased cooking loss in the thigh. Malondialdehyde (MDA) concentration in the thigh was linearly (P=0.020) and quadratically (P=0.017) reduced by ZnPal inclusion. Additionally, ZnPal supplementation tended to linearly enhance total antioxidant capacity (T-AOC) activity of the pectoralis major muscle (P=0.083). The results obtained in the current study indicated that ZnPal inclusion could alter muscular mineral accumulation, improve meat quality, and enhance the muscular antioxidant capacity of broilers, and Zn supplementation in the form of ZnPal at the dosage of 20 mg/kg would be sufficient in improving meat quality and muscular oxidative status.

Synthesis of boronic-acid-functionalized magnetic attapulgite for selective enrichment of nucleosides.

2,4-Difluoro-3-formyl-phenylboronic acid (DFFPBA)-modified magnetic attapulgite (ATP-Fe3O4-NH2-DFFPBA) was synthesized and employed to capture and enrich cis-diol-containing biomolecules. The resulting material exhibited a high saturation magnetization value of 20.71 emu/g, allowing the absorbent to be conveniently magnetically separated. Combining the Fe3O4 nanoparticles with the high specific surface area of attapulgite yielded a material with a high capture capacity (13.78 mg/g) for adenosine. Furthermore, ATP-Fe3O4-NH2-DFFPBA was found to possess remarkable selectivity for adenosine at a low molar ratio of adenosine/2-deoxyadenosine (1:500). The potential applications of this material were explored by using it to extract five nucleosides from urine samples, and the results demonstrate that it can decrease matrix interference and selectively enrich analytes.

Origin and status of homologous proteins of biomineralization (biosilicification) in the taxonomy of phylogenetic domains.

The taxonomic affiliation (in the systematisation of viruses, and biological domains) of known peptides and proteins of biomineralization (silicateins, silaffins, silacidins and silicase) and their primary structure homologues were analyzed (methods in silico; using Uniprot database). The total number of known peptides and proteins of biosilicification was counted. The data of the quantitative distribution of the detected homologues found in nature are presented. The similarity of the primary structures of silaffins, silacidins, silicateins, silicase, and their homologues was 21-94%, 45-98%, 39-50%, and 28-40%, respectively. These homologues are found in many organisms, from the Protista to the higher plants and animals, including humans, as well as in bacteria and extracellular agents, and they perform a variety of biological functions, such as biologically controlled mineralisation. The provisional classification of these biomineralization proteins is presented. The interrelation of the origin of the first organic polymers and biomineralization is discussed.

Impact of cell division on intracellular uptake and nuclear targeting with fluorescent SiC-based nanoparticles.

Semiconductor nanoparticles (NPs) became important and wide-used tool for cell imaging because of their unique remarkable properties. Nevertheless, all previous investigations in this area were done on proliferating cells. For the first time, this work demonstrates strong influence of cell active proliferation/contact inhibition of proliferation on uptake of NPs. In addition, we show that cell division plays key-role in penetration of silicon carbide based NPs (SiC NPs) inside the cell nucleus. This may very likely concern other types of NPs able to reach the cell nuclei. In particular, observed effect of cell division gives perspectives for future selective cancer treatment with NPs.

A comprehensive biological evaluation of ceramic nanoparticles as wear debris.

Patients are exposed internally to nanoparticles (NPs) by wear mechanisms associated with total joint arthroplasty. This tissue-specific retention implies that the biological evaluation of NPs shall be integrative and niche targeting. Here, we report that ceramic zirconia and silicon nitride NPs interfere with MG63 cells' function and remarkably stimulate the secretion of TNF-α in RAW264.7 cells. However, alumina NPs significantly promote the alkaline phosphatase (ALP) activity of MG63 cells at low concentration and do not show irritation to macrophages. In this study, we prove that ceramic materials at nanoscale are bioactive to cells. These findings also suggest that a more rational paradigm for the biosafety evaluation of NPs than is currently in place is needed before their clinical applications. FROM THE CLINICAL EDITOR: In this study, the authors demonstrate that ceramic nano materials associated with normal wear-and-tear of joint prostheses at nanoscale are bioactive to cells.

Effectiveness of serum megakaryocyte potentiating factor in evaluating the effects of chrysotile and its heated products on respiratory organs.

Chrysotile (CH), the most common form of asbestos, is rendered less toxic by heating it at 1000°C and converting it to forsterite (FO-1000). However, further safety tests are needed to evaluate human health risk of these materials. It has been reported that serum concentrations of megakaryocyte potentiating factor N-ERC/mesothelin become elevated in patients with mesotheliomas caused by asbestos exposure. In this study, a single 2mg dose of CH or FO-1000 was intratracheally administered to rats. Within 180days after the administrations, serum N-ERC/mesothelin concentrations, levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in lung tissues and pathological changes in respiratory organs were determined. In the CH group, a significant increase in serum N-ERC/mesothelin concentrations was observed immediately after intratracheal administration, and the elevation lasted for 30days. In lung tissues, positive staining for 8-OHdG in bronchioles, alveolar epithelium, inflammatory cells, and granulomas was evidence of a marked DNA oxidative damage. Furthermore, measurements of 8-OHdG in lung tissues based on the HPLC-ECD method suggested that serum N-ERC/mesothelin concentrations tended to increase when there are significant DNA damages in lung tissues. In contrast, in the FO-1000 group, a marked rise in serum N-ERC/mesothelin concentrations occurred only in the early phase (1-7days) after intratracheal administration. Similarly, FO-1000 induced elevation of 8-OHdG in lung tissues was transient and modest compared with those of the CH-treated animals. In both the CH and FO-1000 groups, we observed significant correlations between serum N-ERC/mesothelin concentrations and lung 8-OHdG concentrations (r=0.559, p=0.001 for the CH group; r=0.516, p=0.01 for the FO-1000 group). In summary, we demonstrated the possibility of using serum N-ERC/mesothelin concentrations as a useful biomarker for early phase exposure to either CH or FO-1000.

Phthalocyanine-polyamine conjugates as pH-controlled photosensitizers for photodynamic therapy.

A series of aryl hydroxyamines prepared by reductive amination were treated with silicon(IV) phthalocyanine dichloride in the presence of pyridine to give the diaxially substituted phthalocyanine-polyamine conjugates 1-5. The electronic absorption, fluorescence emission, and efficiency at generating reactive oxygen species of these compounds were all sensitive to the pH environment. Under acidic conditions, the fluorescence quantum yields and the singlet oxygen quantum yields of these compounds were greatly enhanced in DMF as a result of protonation of the amino moieties, which inhibited the photoinduced electron-transfer deactivation pathway. The Q band was diminished and broadened, and the fluorescence intensity decreased as the pH increased in citrate buffer solutions. The rate of superoxide radical formation was also reduced in a higher pH environment. Compound 3, containing a terminal 4-chlorophenyl group at the axial substituent, showed the most desirable pH-responsive properties, which makes it a promising tumor-selective fluorescence probe and photosensitizer for photodynamic therapy. All of the phthalocyanines 1-5 were highly photocytotoxic against HT29 and HepG2 cells with IC(50) values as low as 0.03 microM. Compound 3 was highly selective toward lysosomes, but not mitochondria of HT29 cells.

Binding, inactivation, and adhesion forces between antimicrobial peptide cecropin P1 and pathogenic E. coli.

The antimicrobial peptide cecropin P1 (CP1) exhibits broad spectrum activity against planktonic bacteria, including Escherichia coli (E. coli). However, its activity when attached to a substrate has not been thoroughly studied. We immobilized CP1 to gold or silicon nitride, and studied how the method of attachment of peptide to the surface affected peptide interaction with and killing of the bacteria. Using the quartz crystal microbalance with dissipation monitoring (QCM-D), we characterized non-specific binding between CP1 to silicon nitride and gold, and covalent binding of cysteine-terminated CP1 (CP1-cys) to gold. The density of CP1-cys adsorbed on gold was more than the density of CP1 on silicon nitride, and activity against E. coli also depended on the method of attachment used to anchor the peptide to the surface. Twelve E. coli strains with known lipopolysaccharide (LPS) structures were studied. Bacterial adhesion with CP1 was strongest for E. coli with long O-antigens, as determined by atomic force microscopy (AFM). This may be caused by CP1 interacting with the hydrophilic part of the LPS, while control bacteria or those with short O-antigens had their hydrophobic lipid A region more exposed. Killing of E. coli due to contact with CP1 was dependent on the method by which the peptide was immobilized. Four out of 12 E. coli strains were killed when contacted with CP1-cys bound to gold via a thiol bond, while all 12 strains could be killed when in contact with CP1 on silicon nitride. In summary, both QCM-D adsorption experiments and adhesion forces measured by AFM showed a relationship between bacteria LPS length and binding or interaction with the antimicrobial peptide, but killing of E. coli by the peptide was most strongly dependent on how the peptide was attached to the surface.

Effects of soluble silicon compound and deep-sea water on biochemical and mechanical properties of bone and the related gene expression in mice.

Silicon has been known as an essential element for bone formation. The silicon contents of sea water increase with increasing of depth: 1.8 ppm Si in deep-sea water (DW) at 612 m in depth versus 0.06 ppm in surface sea water (SW). The effects of soluble silicon (Si) and DW from which NaCl was eliminated were studied in comparison with tap water (TW) and SW in cell cultures and in animal experiments using the control strain of senescence accelerated mouse, SAMR1. Si at 10 ppm as sodium metasilicate or 10% DW in the alpha-MEM medium stimulated cellular viability, marker enzymes of osteoblast and osteoclast cell lines, and the (45)CaCl(2) uptake in those cells in comparison with the medium control. After weanling SAMR1 were maintained for 6 months on a diet containing 200 ppm Si and 39% of DW and SW, DW and Si improved bone biochemical indices such as femoral weight, mineral and collagen content, and marker enzymes of bone formation and resorption as well as mechanical properties as compared to TW. In the femoral bone marrow of SAMR1, the mRNA expression of bone morphogenetic protein-2 (BMP-2), interleukin-11 (IL-11), and runt-related transcription factor 2 (Runx 2), which stimulate osteoblast development as well as type I procollagen (COL1A1) mRNA, were significantly increased in both DW and Si groups. The expressions of both osteoprotegerin (OPG) and receptor activator of NF-kappaB ligand (RANKL) were also elevated, resulting in distinct increases of the OPG/RANKL ratio in both DW and Si groups. The results indicated that a soluble silicate and deep-sea water as its natural material stimulated cell growth in both osteoblasts and osteoclasts in cell culture and promoted bone metabolic turnover in favor of bone formation through stimulation of the related mRNA expression in animal experiments.

Action of K-crown ether on photosystem II electron transport: characterization of the site of action.

We have investigated the inhibitory effect of K-crown (18-crown-6 potassium picrate) on photosystem II (PSII)-enriched membrane fragments and O2-evolving core complexes. K-crown at 2-4 microM inhibits about half the control level of O2-evolution activity in both types of PSII samples. Oxygen-evolution studies demonstrated that the ether works by inactivating the centres and not by interfering with antenna function or energy transfer to the reaction centre. K-crown does not disrupt binding of the extrinsic proteins associated with O2 evolution nor complex with bound Ca2+ or Cl- cofactors, but rather it directly inhibits electron transfer after the tetrameric Mn cluster. Fluorescence studies on active and Tris-treated samples showed that K-crown does not prevent artificial donors from transferring electrons to PSII but like DCMU inhibits on the acceptor side after QA, the primary quinone acceptor. However, the ether is a leaky inhibitor and may also act as a weak donor when the Mn cluster is not present. Oxygen-production experiments using silicomolybdate as an artificial acceptor (which accepts from both pheophytin and QB in PSII membranes) demonstrated that the inhibition is at or near the DCMU site.

Carborundum, a bulk similar to dietary fibers but chemically inert, does not decrease colon carcinogenesis.

Dietary fibers might lower the risk of colorectal cancer, maybe because of their bulking effect. We tested the protection afforded by an inert bulk against carcinogenesis. Thirty rats received an azoxymethane injection and were allocated to a control diet, or to a diet supplemented with 10% carborundum. After 100 days the colons were scored for aberrant crypt foci. Compared to controls, the fecal weight was doubled in carborundum-fed rats (P < 0.001), but the aberrant crypt foci multiplicity was not changed (P = 0.92). The results do not support the hypothesis that intestinal dilution by an inert bulk can protect against colon cancer.