An Optical Fiber Chemical Sensor for the Detection of Copper(II) in Drinking Water.

Highly sensitive plasmonic optical fiber platforms combined with receptors have been recently used to obtain selective sensors. A low-cost configuration can be obtained exploiting a D-shaped plastic optical fiber covered with a multilayer sensing surface. The multilayer consists of a gold film, functionalized with a specific receptor, where the surface plasmon resonance (SPR) occurs. The signal is produced by the refractive index variation occurring as a consequence of the receptor-to analyte binding. In this work, a selective sensor for copper(II) detection in drinking water, exploiting a self-assembled monolayer (SAM) of d,l-penicillamine as the sensing layer, has been developed and tested. Different concentrations of copper(II) in NaCl 0.1 M solutions at different pH values and in a real matrix (drinking water) have been considered. The results show that the sensor is able to sense copper(II) at concentrations ranging from 4 × 10-6 M to 2 × 10-4 M. The use of this optical chemical sensor is a very attractive perspective for fast, in situ and low-cost detection of Cu(II) in drinking water for human health concerns. Furthermore, the possibility of remote control is feasible as well, because optical fibers are employed.

A Mn(II)-Mn(II) center in human prolidase.

Human prolidase, the enzyme responsible for the hydrolysis of the Xaa-Pro/Hyp peptide bonds, is a key player in the recycling of imino acids during the final stage of protein catabolism and extracellular matrix remodeling. Its metal active site composition corresponding to the maximal catalytic activity is still unknown, although prolidase function is of increasing interest due to the link with carcinogenesis and mutations in prolidase gene cause a severe connective tissue disorder. Here, using EPR and ICP-MS on human recombinant prolidase produced in Escherichia coli (hRecProl), the Mn(II) ion organized in a dinuclear Mn(II)-Mn(II) center was identified as the protein cofactor. Furthermore, thermal denaturation, CD/fluorescence spectroscopy and limited proteolysis revealed that the Mn(II) is required for the proper protein folding and that a protein conformational modification is needed in the transition from apo- to Mn(II)loaded-enzyme. The collected data provided a better knowledge of the human holo-prolidase and, although limited to the recombinant enzyme, the exact identity and organization of the metal cofactor as well as the conformational change required for activity were proven.

Solid-phase extraction of PFOA and PFOS from surface waters on functionalized multiwalled carbon nanotubes followed by UPLC-ESI-MS.

This is the first report on the analytical application of multiwalled carbon nanotubes (MWCNTs) as solid-phase extraction (SPE) sorbents for determination in surface waters, at the nanograms per litre level, of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS), the two predominant contaminants among the perfluorinated compounds detected. After the preconcentration step, the quantification was achieved by ultraperformance liquid chromatography-electrospray ionization mass spectrometry. To increase the extraction efficiency towards these amphiphilic compounds, MWCNTs were derivatized with amino-terminated alkyl chains, thus producing a mixed-mode material (MWCNT-R-NH2) combining hydrophobic affinity and anion-exchange properties. Experiments with distilled, tap and river water (pH 3) spiked at different concentrations (10, 15, 30, 100, 200 and 500 ng L(-1)) provided absolute recoveries in the range 71-102% (n = 3, relative standard deviations less than 10%). Analytes were eluted in a single fraction with 6 mL methanol (3 × 10(-4) M NaOH). The within-laboratory reproducibility of the MWCNT-R-NH2 SPE sorbent was evaluated with raw river water, and relative standard deviations less than 15% were obtained (n = 4). Preconcentration factors up to 125 (500-mL sample) made it possible to quantify PFOA and PFOS at low nanograms per litre levels in naturally contaminated river water. The method quantification limits of 10 ng L(-1) for PFOA and 15 ng L(-1) for PFOS were well below the advisory levels for drinking and surface waters. Comparison with non-derivatized MWCNTs highlighted the role of functionalization in improving the adsorption affinity towards these contaminants. MWCNT-R-NH2 maintained their extraction capability for at least eight repeated adsorption/desorption cycles.

Antibacterial activity of glutathione-coated silver nanoparticles against Gram positive and Gram negative bacteria.

In the present paper, we study the mechanism of antibacterial activity of glutathione (GSH) coated silver nanoparticles (Ag NPs) on model Gram negative and Gram positive bacterial strains. Interference in bacterial cell replication is observed for both cellular strains when exposed to GSH stabilized colloidal silver in solution, and microbicidal activity was studied when GSH coated Ag NPs are (i) dispersed in colloidal suspensions or (ii) grafted on thiol-functionalized glass surfaces. The obtained results confirm that the effect of dispersed GSH capped Ag NPs (GSH Ag NPs) on Escherichia coli is more intense because it can be associated with the penetration of the colloid into the cytoplasm, with the subsequent local interaction of silver with cell components causing damages to the cells. Conversely, for Staphylococcus aureus, since the thick peptidoglycan layer of the cell wall prevents the penetration of the NPs inside the cytoplasm, the antimicrobial effect is limited and seems related to the interaction with the bacterial surfaces. Experiments on GSH Ag NPs grafted on glass allowed us to elucidate more precisely the antibacterial mechanism, showing that the action is reduced because of GSH coating and the limitation of the translational freedom of NPs.

Synthesis, characterization and antibacterial activity against Gram positive and Gram negative bacteria of biomimetically coated silver nanoparticles.

In the present work, we describe a simple procedure to produce biomimetically coated silver nanoparticles (Ag NPs), based on the postfunctionalization and purification of colloidal silver stabilized by citrate. Two biological capping agents have been used (cysteine Cys and glutathione GSH). The composition of the capped colloids has been ascertained by different techniques and antibacterial tests on GSH-capped Ag NPs have been conducted under physiological conditions, obtaining values of Minimum Inhibitory Concentration (MIC) of 180 and 15 µg/mL for Staphylococcus aureus and Escherichia coli, respectively. The antibacterial activity of these GSH capped NPs can be ascribed to the direct action of metallic silver NPs, rather than to the bulk release of Ag(+).

GC-MS qualitative analysis of the volatile, semivolatile and volatilizable fractions of soil evidence for forensic application: A chemical fingerprinting.

This paper proposes a GC-MS analytical method that can be used in forensic investigations, for comparative or provenance studies of soils with the aim to reconnect the evidence to their origin. The volatile, semi-volatile and volatilizable compounds present in soil samples of different sources have been extracted and qualitatively analyzed by GC-MS. The different fractions were extracted by ultrasonic assisted extraction (UAE), and the volatilizable compounds were derivatized by BSTFA-TMCS as silylating agent. Sixty-five soil samples from different locations in northern Italy were collected, analyzed and a GC-MS "fingerprint database" has been created in order to easily access the data for the unknown soil sample provenance obtained with the same procedure and GC apparatus. With this purpose, the origin of blind samples, chosen randomly from those collected, was identified based on a qualitative comparison of the MS chromatographic profiles, which obviates the need for quantitative analyses.

Correlation between Elemental Composition/Mobility and Skin Cell Proliferation of Fibrous Nanoclay/Spring Water Hydrogels.

Inorganic hydrogels formulated with spring waters and clay minerals are used to treat musculoskeletal disorders and skin affections. Their underlying mechanism of action for skin disorders is not clear, although it is usually ascribed to the chemical composition of the formulation. The aim of this study was to assess the composition and in vitro release of elements with potential wound healing effects from hydrogels prepared with two nanoclays and natural spring water. In vitro Franz cell studies were used and the element concentration was measured by inductively coupled plasma techniques. Biocompatibility studies were used to evaluate the potential toxicity of the formulation against fibroblasts. The studied hydrogels released elements with known therapeutic interest in wound healing. The released ratios of some elements, such as Mg:Ca or Zn:Ca, played a significant role in the final therapeutic activity of the formulation. In particular, the proliferative activity of fibroblasts was ascribed to the release of Mn and the Zn:Ca ratio. Moreover, the importance of formulative studies is highlighted, since it is the optimal combination of the correct ingredients that makes a formulation effective.

An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2.

The osteoinductive property of strontium was repeatedly proven in the last decades. Compelling in vitro data demonstrated that strontium hydroxyapatite nanoparticles exert a dual action, by promoting osteoblasts-driven matrix secretion and inhibiting osteoclasts-driven matrix resorption. Recombinant human bone morphogenetic protein 2 (rhBMP2) is a powerful osteoinductive biologic, used for the treatment of vertebral fractures and critically-sized bone defects. Although effective, the use of rhBMP2 has limitations due its recombinant morphogen nature. In this study, we examined the comparison between two osteoinductive agents: rhBMP2 and the innovative strontium-substituted hydroxyapatite nanoparticles. To test their effectiveness, we independently loaded Gelfoam sponges with the two osteoinductive agents and used the sponges as agent-carriers. Gelfoam are FDA-approved biodegradable medical devices used as delivery system for musculoskeletal defects. Their porous structure and spongy morphology make them attractive in orthopedic field. The abiotic characterization of the loaded sponges, involving ion release pattern and structure investigation, was followed by in vivo implantation onto the periosteum of healthy mice and comparison of the effects induced by each implant was performed. Abiotic analysis demonstrated that strontium was continuously released from the sponges over 28 days with a pattern similar to rhBMP2. Histological observations and gene expression analysis showed stronger endochondral ossification elicited by strontium compared to rhBMP2. Osteoclast activity was more inhibited by strontium than by rhBMP2. These results demonstrated the use of sponges loaded with strontium nanoparticles as potential bone grafts might provide better outcomes for complex fractures. Strontium nanoparticles are a novel and effective non-biologic treatment for bone injuries and can be used as novel powerful therapeutics for bone regeneration.

Self-Assembled Monolayers of Copper Sulfide Nanoparticles on Glass as Antibacterial Coatings.

We developed an easy and reproducible synthetic method to graft a monolayer of copper sulfide nanoparticles (CuS NP) on glass and exploited their particular antibacterial features. Samples were fully characterized showing a good stability, a neat photo-thermal effect when irradiated in the Near InfraRed (NIR) region (in the so called "biological window"), and the ability to release controlled quantities of copper in water. The desired antibacterial activity is thus based on two different mechanisms: (i) slow and sustained copper release from CuS NP-glass samples, (ii) local temperature increase caused by a photo-thermal effect under NIR laser irradiation of CuS NP-glass samples. This behavior allows promising in vivo applications to be foreseen, ensuring a "static" antibacterial protection tailored to fight bacterial adhesion in the critical timescale of possible infection and biofilm formation. This can be reinforced, when needed, by a photo-thermal action switchable on demand by an NIR light.

High Bactericidal Self-Assembled Nano-Monolayer of Silver Sulfadiazine on Hydroxylated Material Surfaces.

Anti-infective surfaces are a modern strategy to address the issue of infection related to the clinical use of materials for implants and medical devices. Nanocoatings, with their high surface/mass ratio, lend themselves to being mono-layered on the material surfaces to release antibacterial molecules and prevent bacterial adhesion. Here, a "layer-by-layer" (LbL) approach to achieve a self-assembled monolayer (SAM) with high microbicidal effect on hydroxylated surfaces is presented, exploiting the reaction between a monolayer of thiolic functions on glass/quartz surfaces and a newly synthesized derivative of the well-known antibacterial compound silver sulfadiazine. Using several different techniques, it is demonstrated that a nano-monolayer of silver sulfadiazine is formed on the surfaces. The surface-functionalized materials showed efficient bactericidal effect against both Gram-positive and Gram-negative bacteria. Interestingly, bactericidal self-assembled nano-monolayers of silver sulfadiazine could be achieved on a large variety of materials by simply pre-depositing glass-like SiO2 films on their surfaces.

Comparison of Different Swabs for Sampling Inorganic Gunshot Residue from Gunshot Wounds: Applicability and Reliability for the Determination of Firing Distance.

This study concentrates on samples of bare pork skin, with and without bristles, and dried bovine ribs shot with a semi-automatic pistol to find the best methodology and sampling surface in the search for inorganic gunshot residues (IGSR). Four quadrants of known surface areas were sampled at different distances from the bullet's hole with different swabs: tapes in graphite, Leukosilk® white tape, 3M® transparent tape, and a cotton swab to assess the technique able to collect the highest amounts of IGSR with the lowest contribution of the blank. The cotton swab wet in 10% HNO3 gave the best results. The highest amounts of IGSR, measured by ICP-OES and MS, were detected on a surface of 3-cm radius from the bullet's edge. The amount of metals collected decreased with the firing distance between 20 and 60 cm. The procedure was efficient for sampling different tissues like skin and bones.

Selective sequential dissolution for the determination of inorganic indium compounds in the particulate matter of emissions and workplace air.

A speciation method was developed for the determination of inorganic indium compounds in the particulate matter of emissions and from the workplace, based on selective sequential extractions. The main inorganic indium compounds that are expected to be present in the atmospheric particulate matter involving the industrial production of indium and in the manufacture of indium compounds were separated and determined. The procedure has been tested on standard reference materials. The indium recovery was in the range 93 - 105%.

Indium/Gallium Maltolate Effects on Human Breast Carcinoma Cells: In Vitro Investigation on Cytotoxicity and Synergism with Mitoxantrone.

In this study, we aimed to investigate in vitro whether the synthetized indium maltolate (InMal) and gallium maltolate (GaMal) could exert either a toxic effect toward breast cancer cell line MDA-MB-231 or an agonistic activity with mitoxantrone (MTX) in comparison to fibroblast cell line NIH-3T3. Both GaMal and InMal reduced viability of MDA-MB-231, and at a lesser extent of NIH3-T3, in a dose- and time-dependent mode, the outcome was more effective in comparison to MTX sole exposure. Both GaMal and InMal toxicity was reverted by iron citrate addition on NIH3-T3, not on MDA-MB-231, showing indirectly that gallium and indium's mechanisms of action may include iron targeting. The agonistic activity against MDA-MB-231 survival was shown pretreating with 100 µM InMal for 24 h followed by medium exchange with MTX at 10 ng mL-1 or vice-versa but not with co-incubation of both compounds. In particular, InMal pretreating resulted more protective to MTX subsequent exposure.

Author Correction: Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3.

The previously published version of this Article contained an error in Figure 3. In panel a, the residues His667 and Asp669 were incorrectly labelled as His627 and Asp629. The error has been corrected in both the PDF and HTML versions of the Article.

Molecular architecture of the multifunctional collagen lysyl hydroxylase and glycosyltransferase LH3.

Lysyl hydroxylases catalyze hydroxylation of collagen lysines, and sustain essential roles in extracellular matrix (ECM) maturation and remodeling. Malfunctions in these enzymes cause severe connective tissue disorders. Human lysyl hydroxylase 3 (LH3/PLOD3) bears multiple enzymatic activities, as it catalyzes collagen lysine hydroxylation and also their subsequent glycosylation. Our understanding of LH3 functions is currently hampered by lack of molecular structure information. Here, we present high resolution crystal structures of full-length human LH3 in complex with cofactors and donor substrates. The elongated homodimeric LH3 architecture shows two distinct catalytic sites at the N- and C-terminal boundaries of each monomer, separated by an accessory domain. The glycosyltransferase domain displays distinguishing features compared to other known glycosyltransferases. Known disease-related mutations map in close proximity to the catalytic sites. Collectively, our results provide a structural framework characterizing the multiple functions of LH3, and the molecular mechanisms of collagen-related diseases involving human lysyl hydroxylases.

Assessment of the Effects Exerted by Acid and Alkaline Solutions on Bone: Is Chemistry the Answer?

The treatment of corpses with extremely acid or basic liquids is sometimes performed in criminal contexts. A thorough characterization by chemical analysis may provide further help to macroscopic and microscopic analysis; 63 porcine bone samples were treated with solutions at different pH (1-14) for immersion periods up to 70 days, as well as in extremely acidic sulfuric acid solutions (9 M/18 M) and extremely basic sodium hydroxide. Inductively coupled optical emission spectrometry (ICP-OES)/plasma mass spectrometry (ICP-MS), Fourier transform infrared spectroscopy (FT-IR), energy dispersive X-ray analysis (EDX), X-ray powder diffraction (XRPD), and scanning electron microscopy (SEM) showed that only the sulfuric acid solution 18 M was able to completely dissolve the sample. In addition, chemical analysis allowed to recognize the contact between bone and substances. Hydrated calcium sulfate arose from extreme pH. The possibility of detecting the presence of human material within the residual solution was demonstrated, especially with FT-IR, ICP-OES, and EDX.

Bulk Surfaces Coated with Triangular Silver Nanoplates: Antibacterial Action Based on Silver Release and Photo-Thermal Effect.

A layer of silver nanoplates, specifically synthesized with the desired localized surface plasmon resonance (LSPR) features, was grafted on amino-functionalized bulk glass surfaces to impart a double antibacterial action: (i) the well-known, long-term antibacterial effect based on the release of Ag⁺; (ii) an "on demand" action which can be switched on by the use of photo-thermal properties of silver nano-objects. Irradiation of these samples with a laser having a wavelength falling into the so called "therapeutic window" of the near infrared region allows the reinforcement, in the timescale of minutes, of the classical antibacterial effect of silver nanoparticles. We demonstrate how using the two actions allows for almost complete elimination of the population of two bacterial strains of representative Gram-positive and Gram-negative bacteria.

Determination of the post mortem interval in skeletal remains by the comparative use of different physico-chemical methods: Are they reliable as an alternative to 14C?

The determination of the post-mortem interval (PMI) of skeletal remains is a challenging aspect in the forensic field. Previous studies focused their attention on different macroscopic and morphological aspects but a thorough and complete evaluation of the potential of chemical and physical analyses in this field of research has not been performed. In addition to luminol test and Oxford histology index (OHI) reported in a recent paper, widely spread and accessible methods based on physical aspect and chemical characteristics of skeletal remains have been investigated as potential alternatives to dating by determination of 14C. The investigation was performed on a total of 24 archeological and forensic bone samples with known PMI, with inductively coupled plasma optical emission spectrometer (ICP-OES), inductively coupled plasma quadruple mass spectrometry (ICP-MS), Fourier transform infrared (FT-IR) spectroscopy, energy dispersive X-ray analysis (EDX), powder X-ray diffraction analysis (XRPD) and scanning electron microscopy (SEM). Finally, the feasibility of such alternative methods was discussed. Some results such as carbonates/phosphates ratio from FT-IR, the amounts of organic and inorganic matter by EDX, crystallite sizes with XRPD, and surface morphology obtained by SEM, showed significant trends along with PMI. Though, from a chemical point of view cut-off values and gold-standard methods still present challenges, and rather different techniques together can provide useful information toward the assessment of the PMI of skeletal remains. It is however clear that in a hypothetical flowchart those methods may be placed practically at the same level and a choice should always consider the evaluation of results by each technique, execution times and a costs/benefits relationship.

Fluoride release and uptake abilities of different fissure sealants.

BACKGROUND: The long-term capability of resin sealants and glass ionomer cements to release fluoride is associated to a reduction in pit and fissure caries. The regular use of fluoride varnishes/toothpastes can result in the absorption of fluoride into the sealant. The objective of the present study was to assess the fluoride release/uptake capacities of different fissure sealants. MATERIAL AND METHODS: Three different fissure sealants (Fuji Triage/GC, Fissurit FX/Voco and Grandio Seal/Voco) were examined. Ten discs of each material were prepared. Each disc was incubated with distilled water and then the solution analyzed for diluted for fluoride concentration, using a combination of fluoride electrode (OrionGP 1 S/N 13824, Orion Research Inc, Boston, MA, USA) connected to an expandable ion analyzer (Orion 720A, Orion Research Inc, Boston, MA, USA). Standard curves between 1 and 100 ppm F- were used to calibrate the electrode. Cumulative fluoride release was measured on days 1, 2, 3, 5, 7, 21, 35 and 49, then two different fluoride varnishes/pastes (Profluorid Varnish/Voco, MI Paste Plus/GC), were applied to the sealants tested, and fluoride release (after reuptake) was measured on days 56, 70 and 84. RESULTS: Kruskal Wallis test confirmed significant differences in fluoride release between Fuji Triage/GC and Fissurit FX/Voco and Grandio Seal/Voco from day 1 (P < 0.001). The application of fluoride varnish Profluorid Varnish enhanced the fluoride release for all sealants (P < 0.05). MI Paste Plus enhanced the fluoride release for all sealants except for Fuji Triage/GC (P > 0.05). CONCLUSIONS: The GIC-based sealant (Fuji Triage/GC) released significantly more fluoride than the resin sealants tested. The exposure to the fluoridated varnish (Profluorid Varnish) significantly recharged the sealants tested more than the CPP-ACPF toothpaste (MI Paste Plus). KEY WORDS: Fissure sealants, fluoride release, fluoride uptake, glass ionomer cements.

The application of heat-deproteinization to the morphological study of cortical bone: A contribution to the knowledge of the osteonal structure.

Observation of heat-deproteinized cortical bone specimens in incident light enabled the high definition documentation of the osteonal pattern of diaphyseal Haversian bone. This prompted a study to compare these images with those revealed by polarized light microscopy, carried out either on decalcified or thin, undecalcified, resin-embedded sections. Different bone processing methods can reveal structural aspects of the intercellular matrix, depending on the light diffraction mode: birefringency in decalcified sections can be ascribed to the collagen fibrils orientation alone; in undecalcified sections, to both the ordered layout of collagen and the inorganic phase; in the heat-deproteinized samples, exclusively to the hydroxyapatite crystals aggregation mode. The elemental chemical analysis documented low content of carbon and hydrogen, no detectable levels of nitrogen and significantly higher content of calcium and phosphorus in heat-deproteinized samples, as compared with dehydrated controls. In both samples, the X-ray diffraction (XRD) pattern did not show any significant difference in pattern of hydroxyapatite, with no peaks of any possible decomposition phases. Scanning electron microscopic (SEM) morphology of heat-deproteinized samples could be documented with the fracturing technique facilitated by the bone brittleness. The structure of crystal aggregates, oriented in parallel and with marks of time periods, was documented. Comparative study of deproteinized and undecalcified samples showed that the matrix inorganic phase did not undergo a coarse grain thermal conversion until it reached 500°C, maintaining the original crystals structure and orientation. Incident light stereomicroscopy, combined with SEM analysis of deproteinized bone fractured surfaces, is a new enforceable technique which can be used in morphometric studies to improve the understanding of the osteonal dynamics. Microsc. Res. Tech. 79:691-699, 2016. © 2016 Wiley Periodicals, Inc.