A multivariate analysis of Multiple Myeloma subtype plasma cells.

Trusted methods for identifying different Multiple Myeloma (MM) cells and their biological diversity due to their immunophenotypic variety are often little detailed and difficult to find in literature. In this work, we show that micro-Raman spectroscopy can be used to highlight if there is a certain degree of distinction or correlation between the MM subtype plasmacells in relation to the cluster of differentiation (CD45+/CD38+/CD138-) and (CD45-/CD38+/CD138+). After taking samples from the bone marrow of patients with Multiple Myeloma, the PCs were sorted by flow cytometry, selecting the most common CD of the disease, i.e. CD 45, CD38 and CD138. Some spectral differences are observed comparing the Raman spectra of the two set of samples investigated. To better define in which spectral regions there are greater differences and, therefore, to which biological contributions the changes refers, we also explored the principal component analysis (PCA) of the collected Raman data. The spectral variations between the different sorted cells have been highlighted by plotting loading vectors PC1 and PC2, which shows a net differentiation between the two set of cells. Ultimately, the differences shown by PCA have been associated with the spectral variations observed and explained in terms of changes of proteins and lipid contributions. Thus, the differentiation of Multiple Myeloma subtype plasma cells by confocal micro-Raman spectroscopy can be proposed as a diagnostic tool in the speeding up of cell identification, assessing the intracellular biochemical changes that take place in cancer cells.

Plastics occurrence in juveniles of Engraulis encrasicolus and Sardina pilchardus in the Southern Tyrrhenian Sea.

We report the presence of microplastics on the external surface and in the gastrointestinal tract of white late-larval and juvenile stages (fry) of clupeid fishes caught in the Southern Tyrrhenian Sea. The average highest number of plastics debris was recorded on Sardina pilchardus (0.53 items/specimen); a lower average number of items was observed for Engraulis encrasicolus (0.26 items/specimen). The plastics were characterized by fibers that differed in shape, colour and composition. Polyester, polypropylene, polyacrylonitrile, polyethylene, polyamide, nylon, rayon and polyurethane segments were detected by Raman and FTIR spectroscopies. Traces of organic components and dyes, compounds that are generally included in the polymer matrix to modify its base properties, were also identified on microplastics. Our results raise concerns for the potential transfer of synthetic materials through the marine food web and into humans, given the prominent role of S. pilchardus and E. encrasicolus within the food web as main food source for many marine species.

Detection of artificial cellulose microfibers in Boops boops from the northern coasts of Sicily (Central Mediterranean).

Pollution deriving from textile wastes, including industrial and household waste, is recently of great interest due to their environmental impacts. Anthropogenic and synthetic fibers are responsible for negative effects on the quality of water and soil, and, also, their presence damages plant and animal health. In this work, the authors revealed the occurrence of man-made cellulose fibers in specimens of Boops boops from the Northern Sicilian coasts. Bogue was chosen as target species as it has been used as an indicator within the European Marine Strategy Framework Directive (MSFD 2008/56/EC) in order to value the "microplastics status" in the stomach contents. Of the 30 specimens examined, 63.3% of these had ingested fibers items. The number of fibers ranged from 1 to 10 per specimens with an average of 2.7 items/specimen. Fibers length ranged from 0.5 to 30 mm, most of them were black (95%), and a small percentage was red (5%). The ingestion of man-made cellulose fibers, observed for the first time, in Boops boops in the Mediterranean Sea wake-up call and it should attract the attention of the EU for new guidelines where this new type of contaminant is classified harmful as well as plastics.

Flow cytometry and micro-Raman spectroscopy: Identification of hemocyte populations in the mussel Mytilus galloprovincialis (Bivalvia: Mytilidae) from Faro Lake and Tyrrhenian Sea (Sicily, Italy).

Immunological and structural characteristics of hemocyte populations in the mussel Mytilus galloprovincialis (Bivalvia: Mytilidae), going from two different Sicilian habitats (Faro Lake and Tyrrhenian sea), was investigated by means of two different techniques (flow cytometric and micro-Raman spectroscopy analyses). For this purpose, three hundred and sixty mussels Mytilus galloprovincialis were analyzed during November 2017. They were divided into two equal groups (triplicate sample) on the basis of the site of collection (n = 60 caught in Faro Lake - group A, and n = 60 caught in Tyrrhenian Sea - group B). Some several differences between the species of Faro Lake and Tyrrhenian Sea are observed and ascribed to the disruption of immune parameters induced by the variations of some qualitative water parameters (temperature, salinity, dissolved oxygen, pH, ammonium 10, free chlorine, total chlorine, total phosphate, orthofhosphate) recorded in the two habitats. This study is relevant for monitoring the conditions of the sea and Faro Lake, which is strongly influenced by the currents of the Tyrrhenian Sea. Faro lake is well known for the cultivation of mussels and this is part of a coastal habitat of particular interest, consisted of a peculiar biocenotic complex. Further, for the first time, significant different arrangement in the mussels cell structural organization was evidenced by simply following their highly reproducible Raman biomolecular signatures.

Exploring alternative biomarkers of pesticide pollution in clams.

Acetylcholinesterase (AChE) is a reliable biomarker of pesticide exposure although in clams this activity is often very low or undetectable. Carboxylesterases (CEs) exhort several physiological roles, but also respond to pesticides. Searching for an AChE alternative, baseline CE activities were characterised in Ruditapes decussatus gills and digestive glands using five substrates suggestive of different isozymes. The long chain p-nitrophenyl butyrate and 1-naphthyl butyrate were the most sensitive. In the digestive gland, their kinetic parameters (Vmax and Km) and in vitro sensitivity to the organophosphorus metabolite chlorpyrifos oxon (CPX) were calculated. IC50 values, in the pM-nM range, suggest a high protection efficiency of CE-related enzymes towards CPX neurotoxicity. Other targeted enzymes were: activities of glutathione reductase, glutathione peroxidase, catalase, glutathione S-transferases (GSTs) and lactate dehydrogenase in gills and digestive glands. The high GSTs activity and CE/AChE ratio suggests that R. decussatus has a great capacity for enduring pesticide exposure.

Evaluation of biological response induced by molybdenum oxide nanocolloids on in vitro cultured NIH/3T3 fibroblast cells by micro-Raman spectroscopy.

Tailored colloids of uniformly sized and engineered molybdenum oxide nanoparticles were produced, for the first time, by pulsed laser ablation in water. This green technique ensures the formation of contaminant-free nanostructures and the absence of by-products, very useful issues in biological applications. A selective tuning of MoO chemical bonding configurations and a suitable control of nanoparticles size distributions were achieved during the ablation processes by varying the water temperature and by applying an external electric field. The metal redox properties are fundamental factors governing both cell uptake and interaction mode with Mo oxide nanoparticles. Micro-Raman spectroscopy was used to investigate the existence of cellular changes induced by Mo oxide colloids on the fibroblast cell line NIH/3T3 in relation to the molecular vibrations due to proteins, lipids and nucleic acids. The label-free micro-Raman spectroscopy provides an easy and noninvasive method to monitor the harmful effect of toxic agents on cells through ROS production or redox-dependent mechanisms. In view of potential biological applications, molybdenum oxide nanoparticles cytotoxicity towards NIH/3T3 cells was also investigated. A statistical analysis shows that, in the 10-100 µg/mL Mo concentration range, all the colloids are cytotoxic, progressively reducing the cell viability down to 75% upon increasing the concentration. The effect is less pronounced for the oxygen deficient MoO3 samples where cell viability does not fall below 85%. These results open the way to identify potential bioactive products affecting cellular redox status, by using only the Raman spectral data, even before performing lengthy and expensive specific clinical analyses.

Quick stimulation of Alcanivorax sp. by bioemulsificant EPS2003 on microcosm oil spill simulation

Oil spill microcosms experiments were carried out to evaluate the effect of bioemulsificant exopolysaccharide (EPS2003) on quick stimulation of hydrocarbonoclastic bacteria. Early hours of oil spill, were stimulated using an experimental seawater microcosm, supplemented with crude oil and EPS2003 (SW+OIL+EPS2003); this system was monitored for 2 days and compared to control microcosm (only oil-polluted seawater, SW+OIL). Determination of bacterial abundance, heterotrophic cultivable and hydrocarbon-degrading bacteria were carried out. Community composition of marine bacterioplankton was determined by 16S rRNA gene clone libraries. Data obtained indicated that bioemulsificant addition stimulated an increase of total bacterial abundance and, in particular, selection of bacteria related to Alcanivorax genus; confirming that EPS2003 could be used for the dispersion of oil slicks and could stimulate the selection of marine hydrocarbon degraders thus increasing bioremediation process.

Quick stimulation of Alcanivorax sp. by bioemulsificant EPS2003 on microcosm oil spill simulation.

Oil spill microcosms experiments were carried out to evaluate the effect of bioemulsificant exopolysaccharide (EPS2003) on quick stimulation of hydrocarbonoclastic bacteria. Early hours of oil spill, were stimulated using an experimental seawater microcosm, supplemented with crude oil and EPS2003 (SW+OIL+EPS2003); this system was monitored for 2 days and compared to control microcosm (only oil-polluted seawater, SW+OIL). Determination of bacterial abundance, heterotrophic cultivable and hydrocarbon-degrading bacteria were carried out. Community composition of marine bacterioplankton was determined by 16S rRNA gene clone libraries. Data obtained indicated that bioemulsificant addition stimulated an increase of total bacterial abundance and, in particular, selection of bacteria related to Alcanivorax genus; confirming that EPS2003 could be used for the dispersion of oil slicks and could stimulate the selection of marine hydrocarbon degraders thus increasing bioremediation process.

Enantioselective reduction of ketoimines promoted by easily available (S)-proline derivatives.

The behavior of readily synthesized and even commercially available (S)-proline derivatives, was studied in the trichlorosilane-mediated reduction of ketoimines. A small library of structurally and electronically modified chiral Lewis bases was considered; such compounds were shown to promote the enantioselective reduction of different substrates in good chemical yields. In the HSiCl3 addition to the model substrate N-phenylacetophenone imine, the organocatalyst of choice led to the formation of the corresponding amine with good stereoselectivity, up to 75% ee. Theoretical studies were also performed in order to elucidate the origin of the stereoselection.

Triclorosilane-mediated stereoselective synthesis of ß-amino esters and their conversion to highly enantiomerically enriched ß-lactams.

A highly stereoselective trichlorosilane-mediated reduction of N-benzyl enamines was developed; the combination of a low cost, easy to make metal-free catalyst and an inexpensive chiral auxiliary allowed to perform the reaction on substrates with different structural features often with total control of the stereoselectivity. By easy deprotection through hydrogenolysis followed by conversion of ß-aminoester to 2-azetidinones, the synthesis of enantiomerically pure ß-lactams (>98% e.e.) was successfully accomplished.