Microsphere packages of carotenoids: intact sea urchin eggs tracked by Raman spectroscopy tools.

Although natural exposure to ambient UV radiation in oligotrophic seawater at small depths can reach the levels responsible for cellular damage, the sea urchin Paracentrotus lividus is frequently in such sites, particularly on the southern Adriatic Sea shore. Spawning their eggs and spending their early life stage in rocky shores at depths of 0.5-2 m are the results of their successful adaptation strategies, although adults may dwell at greater depths. Surprisingly, there is a paucity of reports regarding the carotenoid content in sea urchin eggs. Beyond their important role in photoprotection against high UV exposure, cell division and early development, the content and distribution of carotenoids contribute to the successful survival of sea urchins and also determine the color of their gonads (roe), which is of commercial importance as a delicacy. Herein, for the first time, we have described the carotenoid content and distribution in intact, freshly released eggs of P. lividus species, non-destructively employing resonance Raman spectroscopy and imaging; near-infrared Raman spectroscopy revealed additional molecular carotenoid content. Echinenone and ß-carotene resonance Raman signals were the most intense, and they were identified as the principal carotenoids that are preferentially accumulated in eggs rather than in gonads. Raman imaging in confocal mode revealed the uniform distribution of the carotenoid signal over the whole eggs, while the distribution of proteins appeared spotted. Egg carotenoids generally maintained their identity after 2 months of dry storage, with slight signs of C[double bond, length as m-dash]C bond oxidation. The potential utilization of P. lividus sea urchin eggs as valuable microsphere packages of native carotenoids is discussed.

A Novel Nanoporous Adsorbent for Pesticides Obtained from Biogenic Calcium Carbonate Derived from Waste Crab Shells.

A novel nanoporous adsorbent was obtained through the thermal treatment and chemical wash of the wasted crab shells (BC1) and characterized by various techniques. The structure of BC1 at the end of the treatments comprised a mixture of calcite and amorphous CaCO3, as evidenced by X-ray diffraction and Fourier transform infrared absorption. The BET surface area, BET pore volume, and pore diameter were 250.33 m2 g-1, 0.4 cm3 g-1, and <70 nm, respectively. The point of zero charge of BC1 was determined to be around pH 9. The prepared adsorbent was tested for its adsorption efficacy towards the neonicotinoid pesticide acetamiprid. The influence of pH (2-10), temperature (20-45 °C), adsorbent dose (0.2-1.2 g L-1), contact time (5-60 min), and initial pesticide concentration (10-60 mg L-1) on the adsorption process of acetamiprid on BC1 was studied. The adsorption capacity of BC1 was 17.8 mg g-1 under optimum conditions (i.e., 20 mg L-1 initial acetamiprid concentration, pH 8, 1 g L-1 adsorbent dose, 25 °C, and 15 min contact time). The equilibrium data obtained from the adsorption experiment fitted well with the Langmuir isotherm model. We developed an effective nanoporous adsorbent for the recycling of crab shells which can be applied on site with minimal laboratory infrastructure according to local needs.

pH-Dependent Behavior of Novel 5-FU Delivery System in Environmental Conditions Comparable to the Gastro-Intestinal Tract.

A biogenic carrier for 5-fluorouracil (5-FU) loading and subsequent tableting as a new drug formulation for slow release has been proposed using the biomineral from blue crab carapace. Due to its highly ordered 3D porous nanoarchitecture, the biogenic carbonate carrier could achieve increased effectiveness in colorectal cancer cure provided that the formulation would successfully pass through the gastric acid conditions. Following the recently proven viability of the concept by demonstrating the slow release of the drug from the carrier using the highly sensitive SERS technique, here we investigated the 5-FU release from the composite tablet drug in pH conditions replicating the gastric environment. The released drug from the tablet was studied in solutions with three relevant pH values, pH 2, pH 3, and pH 4. The 5-FU SERS spectral signature for each pH value was used to build calibration curves for quantitative SERS analysis. The results suggested a similarly slow-releasing pattern in acid pH environments to that in neutral conditions. Although biogenic calcite dissolution was expected in acid conditions, the X-ray diffraction and Raman spectroscopy showed preservation of calcite mineral along with the monohydrocalcite during acid solution exposure for two hours. The total released amount in a time course of seven hours, however, was lower in acidic pH solutions, with a maximum fraction of ~40% of the total amount of loaded drug, for pH 2, as opposed to ~80% for neutral values. Nonetheless, these results clearly prove that the novel composite drug retains its slow-releasing character in environmental conditions compatible with the gastrointestinal pH and that it is a viable and biocompatible alternative for oral delivery of anticancer drug to reach the lower gastro-intestinal tract.

Zinc oxide nanostructures for fluorescence and Raman signal enhancement: a review.

Since the initial discovery of surface-enhanced Raman scattering (SERS) and surface-enhanced fluorescence (SEF), these techniques have shown huge potential for applications in biomedicine, biotechnology, and optical sensors. Both methods rely on the high electromagnetic fields created at locations on the surface of plasmonic metal nanoparticles, depending on the geometry of the nanoparticles, their surface features, and the specific location of analyte molecules. Lately, ZnO-based nanostructures have been exploited especially as SERS substrates showing high enhancement factors and increased charge transfer effect. Additionally, applications focused on enhancing the fluorescence of analyte molecules as well as on tuning the photoluminescence properties of ZnO nanostructures through combination with metal nanoparticles. This review covers the major recent results of ZnO-based nanostructures used for fluorescence and Raman signal enhancement. The broad range of ZnO and ZnO-metal nanostructures synthesis methods are discussed, highlighting low-cost methods and the recyclability of ZnO-based nanosubstrates. Also, the SERS signal enhancement by ZnO-based nanostructures and the influences of lattice defects on the SERS signal are described. The photoluminescence enhancement of ZnO in the presence of noble metal nanoparticles and the molecular fluorescence enhancement in the presence of ZnO alone and in combination with metal nanoparticles are also reviewed.

The Influence of Zn Substitution on Physical Properties of CoFe2O4 Nanoparticles.

Co1−xZnxFe2O4 nanoparticles (0 ≤ x ≤ 1) have been synthesized via a green sol−gel combustion method. The prepared samples were studied using X-ray diffraction measurements (XRD), transmission electron microscopy (TEM), Raman, and magnetic measurements. All samples were found to be single phases and have a cubic Fd-3m structure. EDS analysis confirmed the presence of cobalt, zinc, iron, and oxygen in all studied samples. Raman spectra clearly show that Zn ions are preferentially located in T sites for low Zn concentrations. Due to their high crystallinity, the nanoparticles show high values of the magnetization, which increases with the Zn content for x < 0.5. The magnetic properties are discussed based on Raman results. Co ferrite doped with 30% of Zn produced the largest SAR values, which increase linearly from 148 to 840 W/gMNPs as the H is increased from 20 to 60 kA/m.

Comparative screening the life-time composition and crystallinity variation in gilthead seabream otoliths Sparus aurata from different marine environments.

Differences in crystallinity, structure and composition variation along the growing direction in gilthead seabream, Sparus aurata otoliths that inhabited different environments were determined to evaluate the correlation of spectroscopic and chemical data with the lifetime development and movement pattern. The Raman spectroscopy signal provided the characteristic bands whose Full Width at Half Maximum (FWHM) were used to track the signal variability. The FWHM showed an initial increase in the core area, followed by a decrease depicting two minima coinciding growth rings. The crystal discontinuity linked to annual rings was confirmed. The FWHM pattern followed cycle in the individual's activity. However, no significant correlation with FWHM and environmental factors although the slope of the FWHM variation distinguished aquaculture and costal groups from open sea and transitional, estuarine waters. Raman data were further correlated with morphological and elemental composition obtained via SEM-EDX and by LA-ICP-MS. SEM clearly confirmed CRM findings. Finally, multiparameter analysis of Ba/Ca concentrations obtained by LA-ICP-MS indicated the separation of groups associated with aquaculture and transitional waters due lowest variability in the elemental composition. Other groups are more variable possibly due to the water oligotrophic character and greater variability in prey availability in each environment. Results of the present study showed the additional potential of Raman spectroscopy as a complementary tool for inference of migration or origin of fish based on otolith composition and structure like other well-established technique.

Single-cell Raman micro-spectroscopy for tracking of carotenoids in cyanobacteria exposed to Mn and Zn doped ferrite nanoparticles.

Ferrite nanoparticles are increasingly produced and exploited as adsorbents for environmental pollutants. However, their impact on the aquatic microbiota such as cyanobacteria, are not yet investigated. Targeting the environmental monitoring context, in this paper we explored for the first time if any change in the carotenoid signal from cyanobacteria Coelomoron pussilum (AICB 1012) exposed to non-lethal doses of Mn and Zn doped ferrite nanoparticles (NPs) can be associated with the nano-aggression on single-cell level, using micro-Raman spectroscopy. UV-Vis absorption spectroscopy of the bulk culture and single-cell Raman microscopy showed that the carotenoid signal increases relative to the chlorophyll contribution upon exposure of the cells to the Mn-ferrite NPs throughout the 7 days of the experiment. The red-shift and broadening of the strongest carotenoid Raman band arising from (CC) stretching modes indicates the change of carotenoid profile towards increased amount of ß-carotene in answer to the NPs stress. The increase of this band intensity relative to the fluorescence background was also observed in Zn-ferrite NPs treatment. Using a simplified and rapid sample preparation procedure, electron microscopy in both transmission and scanning modes, showed greater coverage of the cells by the stable colloidal AgNPs than by the magnetic ferrite NPs. The latter mostly clumped together rather than adhering to the cells. The combined single-cell micro-Raman tracking of physiological response of the unicellular photosynthetic microorganisms coupled with electron microscopy approach to visualise cell-NPs interaction and the extracellular polymeric substance secretion holds the promise for rapid assessment of the NPs-induced environmental stress acting on the unicellular organisms.

Promoting hidden natural design templates in wasted shells of the mantis shrimp into valuable biogenic composite.

The mineralized cuticle of the mantis shrimps Squilla Mantis which serve as natural hammers, spears and armors, have attracted research attention from various fields due to its amazing mechanical properties which were studied from evolutionary and ecological points of view. Here we aimed to valorize the astonishing mantis shrimp shell waste resulted from fishery and seafood industry as valuable biogenic composite derived from nature, potentially re-usable for novel, smart materials or added-value by-products, aspect which was not deeply considered before. Employing multi-laser Raman spectroscopy and imaging, supported by x-ray diffraction and high-resolution electron microscopy, we discover that the peripheral segments anatomically known as claws and telson, featured completely different composition and morphology, suggesting different applicability. The claw presents a bulk Mg-CaCO3 structure reinforced with fluorapatite coating, while the carotenoid-rich telson presents a porous and anisotropic structure of an amorphous mixture of CaCO3 and CaPO4 in gradient deposition on the chitin-protein scaffold. Resonance Raman spectroscopy showed concentrated pools of astaxanthin carotenoid within the bright red spots visible on telson, Based on our findings, we discuss this material's potential for selective applicability, as a natural source of phosphate-carbonate minerals, antioxidants, biofertilizer, pollutant adsorbent, valuable material for regenerative medicine or even as a cell culture substrate. Knowledge-based approach on this bio-template is the basis for smart recycling of such fishery waste for sustainable development, by opening channels for blue bioeconomy avenue.

Wasted Biomaterials from Crustaceans as a Compliant Natural Product Regarding Microbiological, Antibacterial Properties and Heavy Metal Content for Reuse in Blue Bioeconomy: A Preliminary Study.

The compliance of crab shells traditionally used as a complex natural product for agricultural soil amendment with modern biofertilizers' quality and safety requirements was investigated. Shells waste from the Blue crab, Callinectes sapidus and the Green crab, Carcinus aestuarii were tested for macronutrients, heavy metals, bacteria content, and antimicrobial properties. Such information is crucial for further utilization of the biogenic powders for any composite formulation in added-value by-products. The calcium carbonate-rich hard tissue yield was 52.13% ± 0.015 (mean ± S.D.) and 64.71% ± 0.144 from the blue and green crabs, respectively. The contents of Pb, Ni, Zn, Cr (VI), and Cu were several orders of magnitude below the prescribed limit by EU biofertilizer legislation, with Fe, Mn (not prescribed), and As being the most abundant. The content of As and Cd from the material considered here was within limits. The shells contain no colony-forming units of Salmonella spp. and compliant levels of Escherichia coli; moreover, the shell micro-powder showed dose-dependent growth inhibition of Pseudomonas aeruginosa and Staphylococcus aureus. In summary, the waste crab shells present a complex natural product as plant biofertilizer following the circular economy concepts.

Novel Drug Carrier: 5-Fluorouracil Formulation in Nanoporous Biogenic Mg-calcite from Blue Crab Shells-Proof of Concept.

The ever-growing demand for novel, cheaper, and more effective drugs has put nanomedicine and targeted drug delivery to the forefront of scientific innovation. Owing to its porous three-dimensional (3D)-nanostructure and properties, the biogenic calcite from wasted blue crab shells is employed in the present work as a new drug carrier for 5-fluorouracil (5-FU), a drug widely used in cancer therapy. The drug solution has been loaded in the porous nanoarchitecture of the powdered biogenic material and further pelleted in tablets with a 5-FU concentration of 1.748 mg/g. Their structural and morphological properties were characterized using Raman, X-ray diffraction, and scanning electron microscopy. Confocal micro-Raman spectra of tablet surface showed a typical signal of biogenic carbonate with preserved carotenoids and carotenoproteins found in the native waste shell, while the drug Raman signal was absent, indicating its adsorption in the intricate nanoporous biogenic carrier. The slow release of the drug from the newly formulated tablet was investigated by tracking the surface-enhanced Raman scattering (SERS) signal of the tablet solution in a series of time-dependent experiments. The SERS signal quantification is achieved using the well-known SERS spectral fingerprint of 5-fluorouracil aqueous solution adsorbed on Ag nanoparticles. The proof of concept is demonstrated by quantifying the slow release of the drug through the characteristic SERS band intensity of 5-FU in a time course of 26 h. This proof of concept boosted further investigations concerning the released drug identity in simulated solutions that mimic the pH of the upper- and lower gastrointestinal tract, as well as the multiple possibilities to control porosity and composition during powdering and treatment of biogenic material, to achieve the most convenient formulation for relevant biomedical drug delivery. Nonetheless, the present results showed great promise for innovative reusing waste biogenic 3D-nanomaterials of aquatic origin as advantageous drug carriers for slow release purposes, in line with the concept of blue bioeconomy.

Biocompatible Magnetic Colloidal Suspension Used as a Tool for Localized Hyperthermia in Human Breast Adenocarcinoma Cells: Physicochemical Analysis and Complex In Vitro Biological Profile.

Magnetic iron oxide nanoparticles are the most desired nanomaterials for biomedical applications due to their unique physiochemical properties. A facile single-step process for the preparation of a highly stable and biocompatible magnetic colloidal suspension based on citric-acid-coated magnetic iron oxide nanoparticles used as an effective heating source for the hyperthermia treatment of cancer cells is presented. The physicochemical analysis revealed that the magnetic colloidal suspension had a z-average diameter of 72.7 nm at 25 °C with a polydispersity index of 0.179 and a zeta potential of -45.0 mV, superparamagnetic features, and a heating capacity that was quantified by an intrinsic loss power analysis. Raman spectroscopy showed the presence of magnetite and confirmed the presence of citric acid on the surfaces of the magnetic iron oxide nanoparticles. The biological results showed that breast adenocarcinoma cells (MDA-MB-231) were significantly affected after exposure to the magnetic colloidal suspension with a concentration of 30 µg/mL 24 h post-treatment under hyperthermic conditions, while the nontumorigenic (MCF-10A) cells exhibited a viability above 90% under the same thermal setup. Thus, the biological data obtained in the present study clearly endorse the need for further investigations to establish the clinical biological potential of synthesized magnetic colloidal suspension for magnetically triggered hyperthermia.

Development and Characterization of Fe3O4@Carbon Nanoparticles and Their Biological Screening Related to Oral Administration.

The current study presents the effect of naked Fe3O4@Carbon nanoparticles obtained by the combustion method on primary human gingival fibroblasts (HGFs) and primary gingival keratinocytes (PGKs)-relevant cell lines of buccal oral mucosa. In this regard, the objectives of this study were as follows: (i) development via combustion method and characterization of nanosized magnetite particles with carbon on their surface, (ii) biocompatibility assessment of the obtained magnetic nanoparticles on HGF and PGK cell lines and (iii) evaluation of possible irritative reaction of Fe3O4@Carbon nanoparticles on the highly vascularized chorioallantoic membrane of a chick embryo. Physicochemical properties of Fe3O4@Carbon nanoparticles were characterized in terms of phase composition, chemical structure, and polymorphic and molecular interactions of the chemical bonds within the nanomaterial, magnetic measurements, ultrastructure, morphology, and elemental composition. The X-ray diffraction analysis revealed the formation of magnetite as phase pure without any other secondary phases, and Raman spectroscopy exhibit that the pre-formed magnetic nanoparticles were covered with carbon film, resulting from the synthesis method employed. Scanning electron microscopy shown that nanoparticles obtained were uniformly distributed, with a nearly spherical shape with sizes at the nanometric level; iron, oxygen, and carbon were the only elements detected. While biological screening of Fe3O4@Carbon nanoparticles revealed no significant cytotoxic potential on the HGF and PGK cell lines, a slight sign of irritation was observed on a limited area on the chorioallantoic membrane of the chick embryo.

Rapid and Application-Tailored Assessment Tool for Biogenic Powders from Crustacean Shell Waste: Fourier Transform-Infrared Spectroscopy Complemented with X-ray Diffraction, Scanning Electron Microscopy, and Nuclear Magnetic Resonance Spectroscopy.

Due to their chemical composition, richness in calcium carbonate, chitin, proteins, and pigments, and nanoporous structure, crustacean shell waste shows great potential for a wide variety of applications. Large quantities of waste shells are produced annually, meaning that they can be considered a renewable source of ecofriendly biogenic materials, which can be turned into value-added byproducts. In this paper, an IR-based technique is developed to differentiate various biogenic powders originated from crude or food-processed crustacean shells. The validity of the method is supported by cross-checking with XRD, NMR, and SEM-EDX analyses. Our goal was to determine changes in properties of waste crab shells after the two most common treatments, deproteinization and milling. We discovered that deproteinization with NaOH could be tracked from the IR absorbance intensity ratio of the υ(CH2,3) and υasym(CO3 2-) bands while milling time less influenced this ratio but induced changes in powder particle size distribution and morphology. The relative organic/inorganic ratio was different for different colored shells. Unexpectedly, waste shells stored for an average of 6 months or more were found to contain hydrated calcium carbonate (monohydrocalcite), which was absent in equivalent fresh shell composition. Deproteinization caused changes in mechanical properties of shells, making them more brittle, which resulted in a larger fraction of fine particles after powdering.

Color-specific porosity in double pigmented natural 3d-nanoarchitectures of blue crab shell.

3D-engineered nano-architectures with various functionalities are still difficult to obtain and translate for real-world applications. However, such nanomaterials are naturally abundant and yet wasted, but could trigger huge interest for blue bioeconomy, provided that our understanding of their ultrastructure-function is achieved. To date, the Bouligand pattern in crustaceans shell structure is believed to be unique. Here we demonstrated that in blue crab Callinectes sapidus, the 3D-nanoarchitecture is color-specific, while the blue and red-orange pigments interplay in different nano-sized channels and pores. Thinnest pores of about 20 nm are found in blue shell. Additionally, the blue pigment co-existence in specific Bouligand structure is proved for the green crab Carcinus aestuarii, although the crab does not appear blue. The pigments interplay, simultaneously detected by Raman spectroscopy in color-specific native cuticles, overturns our understanding in crustaceans coloration and may trigger the selective use of particular colored natural nanoarchitectures for broaden area of applications.

Citrus fruits freshness assessment using Raman spectroscopy.

The freshness of citrus fruits commonly available in the market was non-destructively assessed by Raman spectroscopy. Intact clementine, mandarin and tangerine species were characterised concerning their carotenoids skin Raman signalling in a time course from the moment they were acquired as fresh stock, supplying the market, to the physical degradation, when they were no longer attractive to consumers. The freshness was found to strongly correlate to the peel Raman signal collected from the same area of the intact fruits in a time course of a maximum of 20days. We have shown that the intensity of the carotenoid Raman signal is indeed a good indicator of fruit freshness and introduced a Raman coefficient of freshness (CFresh), whose time course is linearly decreasing, with different slope for different citrus groups. Additionally, we demonstrated that the freshness assessment could be achieved using a portable Raman instrument. The results could have a strong impact for consumer satisfaction and the food industry.