Through tissue imaging of a live breast cancer tumour model using handheld surface enhanced spatially offset Resonance Raman Spectroscopy

In order to improve patient survival and reduce the amount of unnecessary and traumatic biopsies, non-invasive detection of cancerous tumours is of imperative and urgent need. Multicellular tumour spheroids (MTS) can be used as an ex vivo cancer tumour model, to model in vivo nanoparticle (NP) uptake by the enhanced permeability and retention (EPR) effect. Surface enhanced spatially offset Raman spectroscopy (SESORS) combines both surface enhanced Raman spectroscopy (SERS) and spatially offset Raman spectroscopy (SORS) to yield enhanced Raman signals at much greater sub-surface levels. By utilizing a reporter that has an electronic transition in resonance with the laser frequency, surface enhanced resonance Raman scattering (SERRS) yields even greater enhancement in Raman signal. Using a handheld SORS spectrometer with back scattering optics, we demonstrate the detection of live breast cancer 3D multicellular tumour spheroids (MTS) containing SERRS active NPs through 15 mm of porcine tissue. False color 2D heat intensity maps were used to determine tumour model location. In addition, we demonstrate the tracking of SERRS-active NPs through porcine tissue to depths of up to 25 mm. This unprecedented performance is due to the use of red-shifted chalcogenpyrylium-based Raman reporters to demonstrate the novel technique of surface enhanced spatially offset resonance Raman spectroscopy (SESORRS) for the first time. Our results demonstrate a significant step forward in the ability to detect vibrational fingerprints from a tumour model at depth through tissue. Such an approach offers significant promise for the translation of NPs into clinical applications for non-invasive disease diagnostics based on this new chemical principle of measurement

Optimization of electrocatalytic H2O2 production at pilot plant scale for solar-assisted water treatment

This manuscript summarizes the successful start-up and operation of a hybrid eco-engineered water treatmentsystem, at pilot scale. The pilot unit, with 100L capacity, has been devised for the efficient electrocatalyticproduction of H2O2at an air-diffusion cathode, triggering the formation of%OH from Fenton's reaction withadded Fe2+catalyst. These radicals, in combination with those formed at a powerful boron-doped diamond(BDD) anode in an undivided cell, are used to degrade a mixture of model pesticides. The capability of the plantto produce H2O2on site was initially optimized using an experimental design based on central composite design(CCD) coupled with response surface methodology (RSM). This aimed to evaluate the effect of key processparameters like current density (j) and solution pH. The influence of electrolyte concentration as well as liquidand air flow rates on H2O2electrogeneration and current efficiency at optimizedjand pH was also assessed. Thebest operation conditions resulted in H2O2mass production rate of 64.9mgmin−1, 89.3% of current efficiencyand 0.4kWh m-3of energy consumption at short electrolysis time. Performance tests at optimum conditions werecarried out with 75L of a mixture of pesticides (pyrimethanil and methomyl) as a first step towards the elim-ination of organic contaminants by solar photoelectro-Fenton (SPEF) process. The combined action of homo-geneous (%OH) and heterogeneous (BDD(%OH)) catalysis along with photocatalysis (UV photons collected at asolar CPC photoreactor) allowed the removal of more than 50% of both pesticides in 5min, confirming the fastregeneration of Fe2+catalyst through cathodic reduction and photo-Fenton reaction

Membrane and Electrochemical Based Technologies for the Decontamination of Exploitable Streams Produced by Thermochemical Processing of Contaminated Biomass

Phytoremediation is an emerging concept for contaminated soil restoration via the use of resilient plants that can absorb soil contaminants. The harvested contaminated biomass can be thermochemically converted to energy carriers/chemicals, linking soil decontamination with biomass-to-energy and aligning with circular economy principles. Two thermochemical conversion steps of contaminated biomass, both used for contaminated biomass treatment/exploitation, are considered: Supercritical Water Gasification and Fast Pyrolysis. For the former, the vast majority of contaminants are transferred into liquid and gaseous effluents, and thus the application of purification steps is necessary prior to further processing. In Fast Pyrolysis, contaminants are mainly retained in the solid phase, but a part appears in the liquid phase due to fine solids entrainment. Contaminants include heavy metals, particulate matter, and hydrogen sulfide. The purified streams allow the in-process re-use of water for the Super Critical Water Gasification, the sulfur-free catalytic conversion of the fuel-rich gaseous stream of the same process into liquid fuels and recovery of an exploitable bio-oil rich stream from the Fast Pyrolysis. Considering the fundamental importance of purification/decontamination to exploit the aforementioned streams in an integrated context, a review of available such technologies is conducted, and options are shortlisted. Technologies of choice include polymeric-based membrane gas absorption for desulfurization, electrooxidation/electrocoagulation for the liquid product of Supercritical Water Gasification and microfiltration via ceramic membranes for fine solids removal from the Fast Pyrolysis bio-oil. Challenges, risks, and suitable strategies to implement these options in the context of biomass-to-energy conversion are discussed and recommendations are made

Comparative analysis of pSMA198 found in Streptococcus macedonicus ACA-DC 198, the first streptococcal plasmid of the pCI305/pWV02 family of theta-replicating replicons

Here we analyze pSMA198, the first plasmid isolated from Streptococcus macedonicus ACA-DC 198, and we attempt to clarify the route of its original acquisition. Based on the similarity profiles of the plasmid’s replication initiation protein (Rep) and its origin of replication (ori), pSMA198 was found to be a novel member of the pCI305/pWV02 family of theta-replicating plasmids. The pCI305/pWV02 family consists of plasmids of narrow host range that are mainly found in lactococcal species. Comparative analysis of the pSMA198 revealed a high degree of similarity with plasmids pSK11b, pVF22 and pIL5 over its replication backbone, its mobilization backbone and most of its length, respectively. All these three plasmids have been isolated from Lactococcus lactis strains deriving from milk or its products supporting that S. macedonicus acquired pSMA198 from the latter species and that this acquisition took place in the dairy environment. Both pSMA198 and the chromosome of S. macedonicus exhibit a high degree of pseudogenes, indicating that they must have evolved under the same gene decay processes. Furthermore, we were able to determine chromosomal regions that may have originated from pSMA198, also supporting a long co-existence of the two replicons. In addition, pSMA198 is carried by S. macedonicus strains segregated in five different genotypes by pulsed-field gel electrophoresis (PFGE), showing that pSMA198’s acquisition is not a recent event. We propose that our overall analysis of pSMA198 points towards the habituation of S. macedonicus ACA-DC 198 to the dairy environment

Characterization of plasmid pSMA198 found in Streptococcus macedonicus ACA-DC 198 supports the relation of the species to the milk environment

Background: Streptococcus macedonicus is an intriguing streptococcal species whose most frequent source of isolation is fermented foods similarly to Streptococcus thermophilus. During the genome sequencing of S. macedonicus ACA-DC 198 a plasmid was identified. Objectives: To analyse pSMA198, the first plasmid isolated from S. macedonicus and to shed light onto its acquisition path. Methods: Similarity searches of nucleotide and protein sequences, comparative analysis of whole plasmid sequences and phylogenetic analysis were performed using the appropriate bioinformatics tools. Methods: Based on the similarity profiles of the plasmid’s replication initiation protein (Rep) and its origin of replication (ori), pSMA198 belongs to the narrow host range pCI305/pWV02 family found primarily in lactococci and it is the first such plasmid to be reported in streptococci. Comparative analysis of the pSMA198 over its ori, origin of transfer (oriT) or entire length revealed a high degree of similarity with plasmids pSK11b, pVF22 and pIL5, respectively, all isolated from Lactococcus lactis strains from milk or milk products. Phylogenetic analysis of the pSMA198 Rep showed that the vast majority of closely related proteins derive from lactococcal dairy isolates. Conclusions: Our findings demonstrate that S. macedonicus ACA-DC 198 acquired most probably plasmid pSMA198 from L. lactis during an ancestral genetic exchange event that took place in milk or dairy products. Based on our analysis we provide the first molecular and evolutionary evidence for the habituation of S. macedonicus to the dairy environment

Acquisition through Horizontal Gene Transfer of Plasmid pSMA198 by Streptococcus macedonicus ACA-DC 198 Points towards the Dairy Origin of the Species

Background: Streptococcus macedonicus is an intriguing streptococcal species whose most frequent source of isolation is fermented foods similarly to Streptococcus thermophilus. However, S. macedonicus is closely related to commensal opportunistic pathogens of the Streptococcus bovis/Streptococcus equinus complex. Methodology/Principal Findings: We analyzed the pSMA198 plasmid isolated from the dairy strain Streptococcus macedonicus ACA-DC 198 in order to provide novel clues about the main ecological niche of this bacterium. pSMA198 belongs to the narrow host range pCI305/pWV02 family found primarily in lactococci and to the best of our knowledge it is the first such plasmid to be reported in streptococci. Comparative analysis of the pSMA198 sequence revealed a high degree of similarity with plasmids isolated from Lactococcus lactis strains deriving from milk or its products. Phylogenetic analysis of the pSMA198 Rep showed that the vast majority of closely related proteins derive from lactococcal dairy isolates. Additionally, cloning of the pSMA198 ori in L. lactis revealed a 100% stability of replication over 100 generations. Both pSMA198 and the chromosome of S. macedonicus exhibit a high percentage of potential pseudogenes, indicating that they have co-evolved under the same gene decay processes. We identified chromosomal regions in S. macedonicus that may have originated from pSMA198, also supporting a long co-existence of the two replicons. pSMA198 was also found in divergent biotypes of S. macedonicus and in strains isolated from dispersed geographic locations (e.g. Greece and Switzerland) showing that pSMA198’s acquisition is not a recent event. Conclusions/Significance: Here we propose that S. macedonicus acquired plasmid pSMA198 from L. lactis via an ancestral genetic exchange event that took place most probably in milk or dairy products. We provide important evidence that point towards the dairy origin of this species

Multiplex imaging of live breast cancer tumour models through tissue using handheld surface enhanced spatially offset resonance Raman spectroscopy (SESORRS)

Through utilizing the depth penetration capabilities of SESORS, multiplexed imaging and classification of three singleplex nanotags and a triplex of nanotags within breast cancer tumour models is reported for the first time through depths of 10 mm using a handheld SORS instrument

Towards establishing a minimal nanoparticle concentration for applications involving surface enhanced spatially offset resonance Raman spectroscopy (SESORRS) in vivo

Resonant chalcogenpyrylium nanotags demonstrate an exceptional surface enhanced Raman scattering (SERS) performance for use in SORS applications. Using surface enhanced spatially offset Raman spectroscopy (SESORS), nanotags modified with a chalcogenpyrylium dye were observed at concentrations as low as 1 pM through 5 mm of tissue. Calculated limits of detection suggest that these SERS nanotags can be detected at 104 fM using surface enhanced spatially offset resonance Raman scattering (SESORRS) demonstrating their potential for in vivo applications