A novel effective bio-originated methylene blue adsorbent: the porous biosilica from three marine diatom strains of Nanofrustulum spp. (Bacillariophyta).

In the present paper, for the first time the ability of the porous biosilica originated from three marine diatom strains of 'Nanofrustulum spp.' viz. N. wachnickianum (SZCZCH193), N. shiloi (SZCZM1342), N. cf. shiloi (SZCZP1809), to eliminate MB from aqueous solutions was investigated. The highest biomass was achieved under silicate enrichment for N. wachnickianum and N. shiloi (0.98 g L-1 DW and 0.93 g L-1 DW respectively), and under 15 °C for N. cf. shiloi (2.2 g L-1 DW). The siliceous skeletons of the strains were purified with hydrogen peroxide and characterized by SEM, EDS, the N2 adsorption/desorption, XRD, TGA, and ATR-FTIR. The porous biosilica (20 mg DW) obtained from the strains i.e. SZCZCH193, SZCZM1342, SZCZP1809, showed efficiency in 77.6%, 96.8%, and 98.1% of 14 mg L-1 MB removal under pH 7 for 180 min, and the maximum adsorption capacity was calculated as 8.39, 19.02, and 15.17 mg g-1, respectively. Additionally, it was possible to increase the MB removal efficiency in alkaline (pH = 11) conditions up to 99.08% for SZCZP1809 after 120 min. Modelling revealed that the adsorption of MB follows Pseudo-first order, Bangham's pore diffusion and Sips isotherm models.

Removal of the Basic and Diazo Dyes from Aqueous Solution by the Frustules of Halamphora cf. salinicola (Bacillariophyta).

Industrial wastes with hazardous dyes serve as a major source of water pollution, which is considered to have an enormous impact on public health. In this study, an eco-friendly adsorbent, the porous siliceous frustules extracted from the diatom species Halamphora cf. salinicola, grown under laboratory conditions, has been identified. The porous architecture and negative surface charge under a pH of 7, provided by the various functional groups via Si-O, N-H, and O-H on these surfaces, revealed by SEM, the N2 adsorption/desorption isotherm, Zeta-potential measurement, and ATR-FTIR, respectively, made the frustules an efficient mean of removal of the diazo and basic dyes from the aqueous solutions, 74.9%, 94.02%, and 99.81% against Congo Red (CR), Crystal Violet (CV), and Malachite Green (MG), respectively. The maximum adsorption capacities were calculated from isotherms, as follows: 13.04 mg g-1, 41.97 mg g-1, and 33.19 mg g-1 against CR, CV, and MG, respectively. Kinetic and isotherm models showed a higher correlation to Pore diffusion and Sips models for CR, and Pseudo-Second Order and Freundlich models for CV and MG. Therefore, the cleaned frustules of the thermal spring-originated diatom strain Halamphora cf. salinicola could be used as a novel adsorbent of a biological origin against anionic and basic dyes.

Functionalization of Alpha-Lactalbumin by Zinc Ions.

Alpha-lactalbumin (α-LA) and binding of zinc cations to protein were studied. Molecular characteristics of protein was determined by MALDI-TOF/MS and electrophoresis SDS-PAGE, and also, for complexes, it was determined by spectroscopic techniques (ATR-FT-IR and Raman) and microscopic techniques (SEM along with an EDX detector and also TEM). The pH dependence of zeta potential of α-LA was determined in saline solution. The zinc binding to the protein mechanism was investigated; zinc binding to protein kinetics, the molecular modeling by the DFT method, and electron microscopy (SEM and TEM) for microstructure observation were performed. The experiments performed indicate a quick binding process (equilibrium takes place after 2 min of incubation) which occurs onto the surface of α-LA. Zinc cations change the conformation of the protein and create spherical particles from the morphological point of view. DFT studies indicate the participation of acidic functional groups of the protein (aspartic acid and glutamic acid residues), and these have a decisive influence on the interaction with zinc cations. Application studies of general toxicity and cytotoxicity and bioavailability were conducted.

Study on Biogenic Spindle-Shaped Iron-Oxide Nanoparticles by Pseudostaurosira trainorii in Field of Laser Desorption/Ionization Applications.

Nanostructures-assisted laser desorption/ionization mass spectrometry (NALDI-MS) is gaining attention for the analysis of a wide range of molecules. In this present investigation, Pseudostaurosira trainorii mediated biosynthesized iron-oxide nanoparticles (IONPs) have been utilized as nanostructures assisting ionization and desorption for laser desorption/ionization mass spectrometry (LDI-MS). The chain forming diatom, P. trainorii showed efficiency in the production of IONPs against 0.01 M Fe+3 (pH 2) aqueous solution at the intracellular and extracellular level. The whole biomass and external media turned dark orange in color after 3 days of reaction with Fe3+ solution. Scanning electron microscopic (SEM) images illustrated that the surface of Fe3+ exposed frustules of P. trainorii were entirely covered by synthesized nanostructures contrasting with the natural surface ornamentation of control cells. The IONPs loaded frustules also exhibited catalytic properties by decolorizing yellow colored nitrophenol after 3 h of reaction. Transmission electron microscopic (TEM) images confirmed that the produced particles are spindle-shaped with ~50-70 nm length and ~10-30 nm width. The biogenic IONPs were utilized as an inorganic matrix in LDI-MS and showed high sensitivity towards small molecules as glucose, alanine and triacylglycerols at nano- and picomolar level per spot, respectively. The presented biocompatible technique offers new perspectives in nanobiotechnology for the production of spindle-shaped IONPs that can be applied in future for the preparation of NALDI plates.

Synthesis, Physicochemical Characterization, and Antibacterial Performance of Silver-Lactoferrin Complexes.

Antibiotic-resistant bacteria pose one of the major threats to human health worldwide. The issue is fundamental in the case of chronic wound treatment. One of the latest trends to overcome the problem is the search for new antibacterial agents based on silver. Thus, the aim of this research was to synthesize the silver-lactoferrin complex as a new generation of substances for the treatment of infected wounds. Moreover, one of the tasks was to investigate the formation mechanisms of the respective complexes and the influence of different synthesis conditions on the features of final product. The batch-sorption study was performed by applying the Langmuir and Freundlich isotherm models for the process description. Characterization of the complexes was carried out by spectroscopy, spectrometry, and separation techniques, as well as with electron microscopy. Additionally, the biological properties of the complex were evaluated, i.e., the antibacterial activity against selected bacteria and the impact on L929 cell-line viability. The results indicate the formation of a heterogeneous silver-lactoferrin complex that comprises silver nanoparticles. The complex has higher antibacterial strength than both native bovine lactoferrin and Ag+, while being comparable to silver toxicity.

Silver Nanostructured Substrates in LDI-MS of Low Molecular Weight Compounds.

Mass spectrometric techniques can provide data on the composition of a studied sample, utilizing both targeted and untargeted approaches to solve various research problems. Analysis of compounds in the low mass range has practical implications in many areas of research and industry. Laser desorption ionization techniques are utilized for the analysis of molecules in a low mass region using low sample volume, providing high sensitivity with low chemical background. The fabrication of substrates based on nanostructures to assist ionization with well-controlled morphology may improve LDI-MS efficiency for silver nanoparticles with plasmonic properties. In this work, we report an approach for the preparation of silver nanostructured substrates applied as laser desorption ionization (LDI) plates, using the chemical vapor deposition (CVD) technique. Depending on the mass of used CVD precursor, the approach allowed the synthesis of LDI plates with tunable sensitivity for various low molecular weight compounds in both ion-positive and ion-negative modes. Reduced chemical background and sensitivity to small biomolecules of various classes (fatty acids, amino acids and water-soluble metabolites) at nanomolar and picomolar detection levels for lipids such as triacylglycerols, phosphatidylethanolamines and lyso-phosphatidylcholines represent an emerging perspective for applications of LDI-MS plates for the collection of molecular profiles and targeted analysis of low molecular weight compounds for various purposes.

Synthesis and physicochemical characterization of bovine lactoferrin supersaturated complex with iron (III) ions.

The aim of the study was to investigate the process of Fe3+ binding to bLTF. Moreover, the physicochemical characterization of the respective supersaturated complex was studied. The knowledge should be important for the description of processes that may take place in dairy products fortified with iron. Additionally, the synthesized complex can be utilized as a dietary supplement for the treatment of iron deficiency anemia (IDA). Finally, it was shown that formation of supersaturated iron-protein structures which include LTF often accompanies development of neurodegenerative diseases such as Alzheimer or Parkinson. Thus, the study can reveal some aspects of its pathogenesis process. The methodology of the investigation comprised the utilization of batch sorption study and applying Freundlich and Langmuir models. The complex also was characterized by numerous techniques: spectrometric (ICP-MS), spectroscopic (UV-Vis, ATR-FTIR), electron microscopy (TEM-EDX), SDS-PAGE. Based on obtained results the potential mechanisms of iron interaction with protein were described. Moreover, the molecular docking was applied to visualize possible metal binding sites. The respective complex contains ≈ 33.0 mg/g of iron which is nearly 50 Fe3+ per one protein molecule. The cytotoxicity of the obtained complex was evaluated by MTT reduction and LDH release assays on Caco-2 and nL929 cell lines.

Nanostructured Layer of Silver for Detection of Small Biomolecules in Surface-Assisted Laser Desorption Ionization Mass Spectrometry.

A facile approach for the synthesis of a silver nanostructured layer for application in surface-assisted laser desorption/ionization mass spectrometry of low-molecular-weight biomolecules was developed using electrochemical deposition. The deposition was carried out using the following silver salts: trifluoroacetate, acetate and nitrate, varying the voltage and time. The plate based on trifluoroacetate at 10 V for 15 min showed intense SALDI-MS responses for standards of various classes of compounds: fatty acids, cyclitols, saccharides and lipids at a concentration of 1 nmol/spot, with values of the signal-to-noise ratio ≥50. The values of the limit of detection were 0.71 µM for adonitol, 2.08 µM for glucose and 0.39 µM for palmitic acid per spot. SEM analysis of the plate showed anisotropic flower-like microstructures with nanostructures on their surface. The reduced chemical background in the low-mass region can probably be explained by the absence of stabilizers and reducing agents during the synthesis. The plate synthesized with the developed approach showed potential for future use in the analysis of low-molecular-weight compounds of biological relevance. The absence of the need for the utilization of sophisticated equipment and the synthesis time (10 min) may benefit large-scale applications of the layer for the detection of various types of small biomolecules.

The Study of Protein-Cyclitol Interactions.

Investigation of interactions between the target protein molecule and ligand allows for an understanding of the nature of the molecular recognition, functions, and biological activity of protein-ligand complexation. In the present work, non-specific interactions between a model protein (Bovine Serum Albumin) and four cyclitols were investigated. D-sorbitol and adonitol represent the group of linear-structure cyclitols, while shikimic acid and D-(-)-quinic acid have cyclic-structure molecules. Various analytical methods, including chromatographic analysis (HPLC-MS/MS), electrophoretic analysis (SDS-PAGE), spectroscopic analysis (spectrofluorimetry, Fourier transform infrared spectroscopy, and Raman spectroscopy), and isothermal titration calorimetry (ITC), were applied for the description of protein-cyclitol interactions. Additionally, computational calculations were performed to predict the possible binding places. Kinetic studies allowed us to clarify interaction mechanisms that may take place during BSA and cyclitol interaction. The results allow us, among other things, to evaluate the impact of the cyclitol's structure on the character of its interactions with the protein.

Synthesis and physicochemical characterization of zinc-lactoferrin complexes.

One trend of the modern world is the search for new biologically active substances based on renewable resources. Milk proteins can be a solution for such purposes as they have been known for a long time as compounds that can be used for the manufacturing of multiple food and non-food products. Thus, the goal of the work was to investigate the parameters of Zn-bovine lactoferrin (bLTF) interactions, which enables the synthesis of Zn-rich protein complexes. Zinc-bLTF complexes can be used as food additives or wound-healing agents. Methodology of the study included bLTF characterization by sodium dodecyl sulfate-PAGE, MALDI-TOF, and MALDI-TOF/TOF mass spectrometry as well Zn-bLTF interactions by attenuated total reflection-Fourier-transform infrared, Raman spectroscopy, scanning and transmission microscopy, and zeta potential measurements. The obtained results revealed that the factors that affect Zn-bLTF interactions most significantly were found to be pH and ionic strength of the solution and, in particular, the concentration of Zn2+. These findings imply that these factors should be considered when aiming at the synthesis of Zn-bLTF metallocomplexes.

Silver nanoparticles: Synthesis, investigation techniques, and properties.

The unique silver properties, especially in the form of nanoparticles (NPs), allow to utilize them in numerous applications. For instance, Ag NPs can be utilized for the production of electronic and solar energy harvesting devices, in advanced analytical techniques (NALDI, SERS), catalysis and photocatalysis. Moreover, the Ag NPs can be useful in medicine for bioimaging, biosensing as well as in antibacterial and anticancer therapies. The Ag NPs utilization requires comprehensive knowledge about their features regarding the synthesis approaches as well as exploitation conditions. Unfortunately, a large number of scientific articles provide only restricted information according to the objects under investigation. Additionally, the results could be affected by artifacts introduced with exploited equipment, the utilized technique or sample preparation stages. However, it is rather difficult to get information about problems, which may occur during the studies. Thus, the review provides information about novel trends in the Ag NPs synthesis, among which the physical, chemical, and biological approaches can be found. Basic information about approaches for the control of critical parameters of NPs, i.e. size and shape, was also revealed. It was shown, that the reducing agent, stabilizer, the synthesis environment, including trace ions, have a direct impact on the Ag NPs properties. Further, the capabilities of modern analytical techniques for Ag NPs and nanocomposites investigations were shown, among other microscopic (optical, TEM, SEM, STEM, AFM), spectroscopic (UV-Vis, IR, Raman, NMR, electron spectroscopy, XRD), spectrometric (MALDI-TOF MS, SIMS, ICP-MS), and separation (CE, FFF, gel electrophoresis) techniques were described. The limitations and possible artifacts of the techniques were mentioned. A large number of presented techniques is a distinguishing feature, which makes the review different from others. Finally, the physicochemical and biological properties of Ag NPs were demonstrated. It was shown, that Ag NPs features are dependent on their basic parameters, such as size, shape, chemical composition, etc. At the end of the review, the modern theories of the Ag NPs toxic mechanism were shown in a way that has never been presented before. The review should be helpful for scientists in their own studies, as it can help to prepare experiments more carefully.

A New Approach for Spontaneous Silver Ions Immobilization onto Casein.

The work presents the kinetic and isotherm studies of silver binding on casein, which was carried out using batch sorption technique. Moreover, the influence of light irradiation on the process was shown. In order to investigate the mechanism of metal ions sorption by casein the zero, pseudo-first order kinetics and Weber-Morris intra-particle diffusion as well as Langmuir and Freundlich isotherm models were used. Furthermore, to specify more precisely, the possible binding mechanism, the spectroscopic (FT-IR-Fourier Transform Infrared Spectroscopy, Raman), spectrometric (MALDI-TOF MS-Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry), microscopic (SEM-Scanning Electron Microscope, TEM/EDX-Transmission Electron Microscopy/Energy Dispersive X-ray detector) and thermal (TGA-Thermogravimetric Analysis, DTG-Derivative Thermogravimetry) analysis were performed. Kinetic study indicates that silver binding onto casein is a heterogeneous process with two main stages: initial rapid stage related to surface adsorption onto casein with immediate creation of silver nanoparticles and slower second stage of intraglobular diffusion with silver binding in chelated form (metalloproteins) or ion-exchange form. Spectroscopic techniques confirmed the binding process and MALDI-TOF MS analysis show the dominant contribution of the α-casein in the process. Moreover, the treatment of silver-casein complex by artificial physiological fluids was performed.