Manipulating solid-state spin concentration through charge transport.

Solid-state defects are attractive platforms for quantum sensing and simulation, e.g., in exploring many-body physics and quantum hydrodynamics. However, many interesting properties can be revealed only upon changes in the density of defects, which instead is usually fixed in material systems. Increasing the interaction strength by creating denser defect ensembles also brings more decoherence. Ideally one would like to control the spin concentration at will while keeping fixed decoherence effects. Here, we show that by exploiting charge transport, we can take some steps in this direction, while at the same time characterizing charge transport and its capture by defects. By exploiting the cycling process of ionization and recombination of NV centers in diamond, we pump electrons from the valence band to the conduction band. These charges are then transported to modulate the spin concentration by changing the charge state of material defects. By developing a wide-field imaging setup integrated with a fast single photon detector array, we achieve a direct and efficient characterization of the charge redistribution process by measuring the complete spectrum of the spin bath with micrometer-scale spatial resolution. We demonstrate a two-fold concentration increase of the dominant spin defects while keeping the T2 of the NV center relatively unchanged, which also provides a potential experimental demonstration of the suppression of spin flip-flops via hyperfine interactions. Our work paves the way to studying many-body dynamics with temporally and spatially tunable interaction strengths in hybrid charge-spin systems.

Multipass wide-field phase imager.

Advances in optical imaging always look for an increase in sensitivity and resolution among other practicability aspects. Within the same scope, in this work we report a versatile interference contrast imaging technique, with high phase sensitivity and a large field-of-view of several mm2. Sensitivity is increased through the use of a self-imaging non-resonant cavity, which causes photons to probe the sample in multiple rounds before being detected, where the configuration can be transmissive or reflective. Phase profiles can be resolved individually for each round thanks to a specially designed single-photon camera with time-of-flight capabilities and true pixels-off gating. Measurement noise is reduced by novel data processing combining the retrieved sample profiles from multiple rounds. Our protocol is especially useful under extremely low light conditions as required by biological or photo-sensitive samples. Results demonstrate more than a four-fold reduction in phase measurement noise, compared to single round imaging, and values close to the predicted sensitivity in case of the best possible cavity configuration, where all photons are maintained until n rounds. We also find good agreement with the theoretical predictions for low number of rounds, where experimental imperfections would play a minor role. The absence of a laser or cavity lock-in mechanism makes the technique an easy to use inspection tool.

Corrigendum: In Living Color: Pigment-Based Microbial Ecology At the Mineral-Air Interface.

[This corrects the article DOI: 10.1093/biosci/biac091.].

Progression after First-Line Cyclin-Dependent Kinase 4/6 Inhibitor Treatment: Analysis of Molecular Mechanisms and Clinical Data.

Cyclin-dependent kinase 4/6 inhibitors (CDK4/6iss) are widely used in first-line metastatic breast cancer. For patients with progression under CDK4/6is, there is currently no standard treatment recommended at the category 1 level in international guidelines. The purpose of this article is to review the cellular mechanisms underlying the resistance to CDK4/6is, as well as treatment strategies and the clinical data about the efficacy of subsequent treatments after CDK4/6is-based therapy. In the first part, this review mainly discusses cell-cycle-specific and cell-cycle-non-specific resistance to CDK4/6is, with a focus on early and late progression. In the second part, this review analyzes potential therapeutic approaches and the available clinical data on them: switching to other CDK4/6is, to another single hormonal therapy, to other target therapies (PI3K, mTOR and AKT) and to chemotherapy.

A robust and versatile platform for image scanning microscopy enabling super-resolution FLIM.

Image scanning microscopy (ISM) can improve the effective spatial resolution of confocal microscopy to its theoretical limit. However, current implementations are not robust or versatile, and are incompatible with fluorescence lifetime imaging (FLIM). We describe an implementation of ISM based on a single-photon detector array that enables super-resolution FLIM and improves multicolor, live-cell and in-depth imaging, thereby paving the way for a massive transition from confocal microscopy to ISM.

In Living Color: Pigment-Based Microbial Ecology At the Mineral-Air Interface.

Pigment-based color is one of the most important phenotypic traits of biofilms at the mineral-air interface (subaerial biofilms, SABs), because it reflects the physiology of the microbial community. Because color is the hallmark of all SABs, we argue that pigment-based color could convey the mechanisms that drive microbial adaptation and coexistence across different terrestrial environments and link phenotypic traits to community fitness and ecological dynamics. Within this framework, we present the most relevant microbial pigments at the mineral-air interface and discuss some of the evolutionary landscapes that necessitate pigments as adaptive strategies for resource allocation and survivability. We report several pigment features that reflect SAB communities' structure and function, as well as pigment ecology in the context of microbial life-history strategies and coexistence theory. Finally, we conclude the study of pigment-based ecology by presenting its potential application and some of the key challenges in the research.

Invasive alien species as an environmental stressor and its effects on coping style in a native competitor, the Eurasian red squirrel.

Free-living animals cope with environmental stressors through physiological and behavioural responses. According to the unidimensional model, these responses are integrated within a coping style: proactive individuals (bold, active-explorative and social) have a lower hypothalamic-pituitary-adrenal (HPA) axis reactivity than reactive ones (shy, less active-explorative, less social). These associations may change when individuals are exposed to human-induced rapid environmental change (HIREC), such as the introduction of invasive alien species (IAS). Here, we studied Eurasian red squirrels to investigate the relationship between personality traits and one integrated measure of HPA axis activity, both in areas uncolonized (natural populations) and colonized by an IAS, the Eastern grey squirrel (invaded populations). We expected an association between physiological and behavioural responses, and that activity, exploration and social tendency would covary, forming a behavioural syndrome in natural populations, while competition with the IAS was predicted to disrupt these associations. We used faecal glucocorticoid metabolites (FGMs) as an integrated measure of adrenocortical activity, and measured the levels of four personality traits (exploration, activity, activity-exploration and social tendency) with an open field test and a mirror image stimulation test. We found no correlation between FGMs and personality traits, neither in natural nor invaded populations. However, we found correlations among personality traits in areas without interspecific competition, indicating a behavioural syndrome, which was disrupted in invaded populations. This is one of the few studies showing that an IAS, acting as an environmental stressor, alters a native species' behavioural syndrome, but does not influence its coping style.

Metabolomic and Proteomic Changes in Candida albicans Biofilm in Response to Zosteric Acid Treatment.

Zosteric acid (ZA) is a secondary metabolite of the seagrass Zostera marina, with antibiofilm activity against fungi. Information concerning its mechanisms of action is lacking and this limits the development of more potent derivatives based on the same target and activity structure. The aim of this work was to investigate the ZA mode of action by analyzing the metabolic status of Candida albicans biofilm and its protein expression profile upon ZA treatment. Fourier-Transform Infrared Spectroscopy confirmed that ZA modified the metabolomic response of treated cells, showing changes in the spectral regions, mainly related to the protein compartment. Nano Liquid Chromatography-High-Resolution Mass Spectrometry highlighted that 10 proteins were differentially expressed in the C. albicans proteome upon ZA treatment. Proteins involved in the biogenesis, structure and integrity of cell walls as well as adhesion and stable attachment of hyphae were found downregulated, whereas some proteins involved in the stress response were found overexpressed. Additionally, ZA was involved in the modulation of non-DNA-based epigenetic regulatory mechanisms triggered by reactive oxygen species. These results partially clarified the ZA mechanism of action against fungi and provided insight into the major C. albicans pathways responsible for biofilm formation.

Single Photon Avalanche Diode Arrays for Time-Resolved Raman Spectroscopy.

The detection of peaks shifts in Raman spectroscopy enables a fingerprint reconstruction to discriminate among molecules with neither labelling nor sample preparation. Time-resolved Raman spectroscopy is an effective technique to reject the strong fluorescence background that profits from the time scale difference in the two responses: Raman photons are scattered almost instantaneously while fluorescence shows a nanoseconds time constant decay. The combination of short laser pulses with time-gated detectors enables the collection of only those photons synchronous with the pulse, thus rejecting fluorescent ones. This review addresses time-gating issues from the sensor standpoint and identifies single photon avalanche diode (SPAD) arrays as the most suitable single-photon detectors to be rapidly and precisely time-gated without bulky, complex, or expensive setups. At first, we discuss the requirements for ideal Raman SPAD arrays, particularly focusing on the design guidelines for optimized on-chip processing electronics. Then we present some existing SPAD-based architectures, featuring specific operation modes which can be usefully exploited for Raman spectroscopy. Finally, we highlight key aspects for future ultrafast Raman platforms and highly integrated sensors capable of undistorted identification of Raman peaks across many pixels.

SPADs and SiPMs Arrays for Long-Range High-Speed Light Detection and Ranging (LiDAR).

Light Detection and Ranging (LiDAR) is a 3D imaging technique, widely used in many applications such as augmented reality, automotive, machine vision, spacecraft navigation and landing. Achieving long-ranges and high-speed, most of all in outdoor applications with strong solar background illumination, are challenging requirements. In the introduction we review different 3D-ranging techniques (stereo-vision, projection with structured light, pulsed-LiDAR, amplitude-modulated continuous-wave LiDAR, frequency-modulated continuous-wave interferometry), illumination schemes (single point and blade scanning, flash-LiDAR) and time-resolved detectors for LiDAR (EM-CCD, I-CCD, APD, SPAD, SiPM). Then, we provide an extensive review of silicon- single photon avalanche diode (SPAD)-based LiDAR detectors (both commercial products and research prototypes) analyzing how each architecture faces the main challenges of LiDAR (i.e., long ranges, centimeter resolution, large field-of-view and high angular resolution, high operation speed, background immunity, eye-safety and multi-camera operation). Recent progresses in 3D stacking technologies provided an important step forward in SPAD array development, allowing to reach smaller pitch, higher pixel count and more complex processing electronics. In the conclusions, we provide some guidelines for the design of next generation SPAD-LiDAR detectors.

Spot Tracking and TDC Sharing in SPAD Arrays for TOF LiDAR.

Light Detection and Ranging (LiDAR) is a widespread technique for 3D ranging and has widespread use in most automated systems that must interact with the external environment, for instance in industrial and security applications. In this work, we study a novel architecture for Single Photon Avalanche Diode (SPAD) arrays suitable for handheld single point rangefinders, which is aimed at the identification of the objects' position in the presence of strong ambient background illumination. The system will be developed for an industrial environment, and the array targets a distance range of about 1 m and a precision of few centimeters. Since the laser spot illuminates only a small portion of the array, while all pixels are exposed to background illumination, we propose and validate through Monte Carlo simulations a novel architecture for the identification of the pixels illuminated by the laser spot to perform an adaptive laser spot tracking and a smart sharing of the timing electronics, thus significantly improving the accuracy of the distance measurement. Such a novel architecture represents a robust and effective approach to develop SPAD arrays for industrial applications with extremely high background illumination.

High concentration factor diffractive microlenses integrated with CMOS single-photon avalanche diode detector arrays for fill-factor improvement.

Large-format single-photon avalanche diode (SPAD) arrays often suffer from low fill-factors-the ratio of the active area to the overall pixel area. The detection efficiency of these detector arrays can be vastly increased with the integration of microlens arrays designed to concentrate incident light onto the active areas and may be refractive or diffractive in nature. The ability of diffractive optical elements (DOEs) to efficiently cover a square or rectangular pixel, combined with their capability of working as fast lenses (i.e., ∼f/3) makes them versatile and practical lens designs for use in sparse photon applications using microscale, large-format detector arrays. Binary-mask-based photolithography was employed to fabricate fast diffractive microlenses for two designs of 32×32 SPAD detector arrays, each design having a different pixel pitch and fill-factor. A spectral characterization of the lenses is performed, as well as analysis of performance under different illumination conditions from wide- to narrow-angle illumination (i.e., f/2 to f/22 optics). The performance of the microlenses presented exceeds previous designs in terms of both concentration factor (i.e., increase in light collection capability) and lens speed. Concentration factors greater than 33× are achieved for focal lengths in the substrate material as short as 190µm, representing a microlens f-number of 3.8 and providing a focal spot diameter of <4µm. These results were achieved while retaining an extremely high degree of performance uniformity across the 1024 devices in each case, which demonstrates the significant benefits to be gained by the implementation of DOEs as part of an integrated detector system using SPAD arrays with very small active areas.

Biometric Signals Estimation Using Single Photon Camera and Deep Learning.

The problem of performing remote biomedical measurements using just a video stream of a subject face is called remote photoplethysmography (rPPG). The aim of this work is to propose a novel method able to perform rPPG using single-photon avalanche diode (SPAD) cameras. These are extremely accurate cameras able to detect even a single photon and are already used in many other applications. Moreover, a novel method that mixes deep learning and traditional signal analysis is proposed in order to extract and study the pulse signal. Experimental results show that this system achieves accurate results in the estimation of biomedical information such as heart rate, respiration rate, and tachogram. Lastly, thanks to the adoption of the deep learning segmentation method and dependability checks, this method could be adopted in non-ideal working conditions-for example, in the presence of partial facial occlusions.

Multispectral Depth-Resolved Fluorescence Lifetime Spectroscopy Using SPAD Array Detectors and Fiber Probes.

Single Photon Avalanche Diode (SPAD) arrays are increasingly exploited and have demonstrated potential in biochemical and biomedical research, both for imaging and single-point spectroscopy applications. In this study, we explore the application of SPADs together with fiber-optic-based delivery and collection geometry to realize fast and simultaneous single-point time-, spectral-, and depth-resolved fluorescence measurements at 375 nm excitation light. Spectral information is encoded across the columns of the array through grating-based dispersion, while depth information is encoded across the rows thanks to a linear arrangement of probe collecting fibers. The initial characterization and validation were realized against layered fluorescent agarose-based phantoms. To verify the practicality and feasibility of this approach in biological specimens, we measured the fluorescence signature of formalin-fixed rabbit aorta samples derived from an animal model of atherosclerosis. The initial results demonstrate that this detection configuration can report fluorescence spectral and lifetime contrast originating at different depths within the specimens. We believe that our optical scheme, based on SPAD array detectors and fiber-optic probes, constitute a powerful and versatile approach for the deployment of multidimensional fluorescence spectroscopy in clinical applications where information from deeper tissue layers is important for diagnosis.

Promoting Beneficial and Inhibiting Undesirable Biofilm Formation with Mangrove Extracts.

The extracts of two mangrove species, Bruguiera cylindrica and Laguncularia racemosa, have been analyzed at sub-lethal concentrations for their potential to modulate biofilm cycles (i.e., adhesion, maturation, and detachment) on a bacterium, yeast, and filamentous fungus. Methanolic leaf extracts were also characterized, and MS/MS analysis has been used to identify the major compounds. In this study, we showed the following. (i) Adhesion was reduced up to 85.4% in all the models except for E. coli, where adhesion was promoted up to 5.10-fold. (ii) Both the sum and ratio of extracellular polysaccharides and proteins in mature biofilm were increased up to 2.5-fold and 2.6-fold in comparison to the negative control, respectively. Additionally, a shift toward a major production of exopolysaccharides was found coupled with a major production of both intracellular and extracellular reactive oxygen species. (iii) Lastly, detachment was generally promoted. In general, the L. racemosa extract had a higher bioactivity at lower concentrations than the B. cylindrica extract. Overall, our data showed a reduction in cells/conidia adhesion under B. cylindrica and L. racemosa exposure, followed by an increase of exopolysaccharides during biofilm maturation and a variable effect on biofilm dispersal. In conclusion, extracts either inhibited or enhanced biofilm development, and this effect depended on both the microbial taxon and biofilm formation step.

Sub-lethal concentrations of Perilla frutescens essential oils affect phytopathogenic fungal biofilms.

The lack of deep knowledge of plant pathogenic fungal biofilms is reflected in the few existing environmental-friendly options for controlling fungal plant disease. Indeed, chemical fungicides still dominate the market but present-day concerns about their real efficacy, increasing awareness of the risk they pose to human health and the environment, and the incidence of fungicide resistance have all led to the current trend of near zero-market-tolerance for pesticide residues in fruit and vegetables. Here, essential oils (PK and PK-IK) from the edible leaves of two cultivars of Perilla frutescens are proposed as new, effective, non-toxic, eco-friendly pesticide-free options suitable for a preventive or integrative approach for sustainable crop protection and product preservation. PK and PK-IK were extracted and characterized, and their ability to affect the biofilm formation of the phytopathogenic model fungi Colletotrichum musae, Fusarium dimerum and Fusarium oxysporum was studied at non-lethal doses. Both essential oils at 1000 and 2000 mg l-1 showed excellent anti-biofilm performance: i) reducing conidia adhesion up to 80.3 ±â€¯16.2%; ii) inhibiting conidia germination up to 100.0 ±â€¯0.0%; iii) affecting biofilm structural development, with a reduction in dry weight of up to 100.0 ±â€¯0.0% and extracellular polysaccharides and proteins up to 81.4 ±â€¯8.0% and 51.0 ±â€¯6.1% respectively. In all cases PK-IK showed better activity than PK.

Recent progress in bio-inspired biofilm-resistant polymeric surfaces.

Any surface of human interest can serve as a substrate for biofilm growth, sometimes with detrimental effects. The social and economic consequences of biofilm-mediated damage to surfaces are significant, the financial impact being estimated to be billions of dollars every year. After describing traditional biocide-based approaches for the remediation of biofilm-affected surfaces, this review deals with more recent developments in material science, focusing on non-toxic, eco-sustainable nature-inspired biomaterials with anti-biofilm properties superior to the conventional biocide-based approaches in terms of addressing the biofilm problem.

Eribulin Treatment in Patients with Liver Metastatic Breast Cancer: Eight Italian Case Reports.

Liver metastases are very common in metastatic breast cancer (MBC); current treatments for these lesions are based on systemic chemotherapy, endocrine- or human epidermal growth factor receptor 2 (HER2)-targeted therapy, and palliative therapy. However, no standard approach has been clearly identified for second and further chemotherapy lines in MBC patients. In the phase III clinical trial EMBRACE, eribulin was particularly effective in reducing liver lesions and improving both overall survival and progression-free survival in liver MBC patients. In this series, we collected 8 case reports of Italian clinical practice in which eribulin has shown significant efficacy in reducing liver metastases in MBC patients: complete response was reported in 2 patients, and 4 patients achieved partial response. The treatment was well tolerated, thus confirming that eribulin is a suitable therapeutic option for elderly patients and for those who have metastatic HER2-negative disease. In the setting of MBC, the sequencing of therapeutic agents should consider expected response, side effects, tumor characteristics, and patient's preferences, in order to successfully tailor the most appropriate therapy beyond earlier lines.

Hindering the formation and promoting the dispersion of medical biofilms: non-lethal effects of seagrass extracts.

BACKGROUND: Biofilms have great significance in healthcare-associated infections owing to their inherent tolerance and resistance to antimicrobial therapies. New approaches to prevent and treat unwanted biofilms are urgently required. To this end, three seagrass species (Enhalus acoroides, Halophila ovalis and Halodule pinifolia) collected in Vietnam and in India were investigated for their effects in mediating non-lethal interactions on sessile bacterial (Escherichia coli) and fungal (Candida albicans) cultures. The present study was focused on anti-biofilm activities of seagrass extracts, without killing cells. METHODS: Methanolic extracts were characterized, and major compounds were identified by MS/MS analysis. The antibiofilm properties of the seagrass extracts were tested at sub-lethal concentrations by using microtiter plate adhesion assay. The performance of the most promising extract was further investigated in elegant bioreactors to reproduce mature biofilms both at the solid/liquid and the solid/air interfaces. Dispersion and bioluminescent assays were carried out to decipher the mode of action of the bioactive extract. RESULTS: It was shown that up to 100 ppm of crude extracts did not adversely affect microbial growth, nor do they act as a carbon and energy source for the selected microorganisms. Seagrass extracts appear to be more effective in deterring microbial adhesion on hydrophobic surfaces than on hydrophilic. The results revealed that non-lethal concentrations of E. acoroides leaf extract: i) reduce bacterial and fungal coverage by 60.9 and 73.9%, respectively; ii) affect bacterial biofilm maturation and promote dispersion, up to 70%, in fungal biofilm; iii) increase luminescence in Vibrio harveyi by 25.8%. The characterization of methanolic extracts showed the unique profile of the E. acoroides leaf extract. CONCLUSIONS: E. acoroides leaf extract proved to be the most promising extract among those tested. Indeed, the selected non-lethal concentrations of E. acoroides leaf extract were found to exert an antibiofilm effect on C. albicans and E. coli biofilm in the first phase of biofilm genesis, opening up the possibility of developing preventive strategies to hinder the adhesion of microbial cells to surfaces. The leaf extract also affected the dispersion and maturation steps in C. albicans and E. coli respectively, suggesting an important role in cell signaling processes.

α-Chymotrypsin Immobilized on a Low-Density Polyethylene Surface Successfully Weakens Escherichia coli Biofilm Formation.

The protease α-chymotrypsin (α-CT) was covalently immobilized on a low-density polyethylene (LDPE) surface, providing a new non-leaching material (LDPE-α-CT) able to preserve surfaces from biofilm growth over a long working timescale. The immobilized enzyme showed a transesterification activity of 1.24 nmol/h, confirming that the immobilization protocol did not negatively affect α-CT activity. Plate count viability assays, as well as confocal laser scanner microscopy (CLSM) analysis, showed that LDPE-α-CT significantly impacts Escherichia coli biofilm formation by (i) reducing the number of adhered cells (-70.7 ± 5.0%); (ii) significantly affecting biofilm thickness (-81.8 ± 16.7%), roughness (-13.8 ± 2.8%), substratum coverage (-63.1 ± 1.8%), and surface to bio-volume ratio (+7.1 ± 0.2-fold); and (iii) decreasing the matrix polysaccharide bio-volume (80.2 ± 23.2%). Additionally, CLSM images showed a destabilized biofilm with many cells dispersing from it. Notably, biofilm stained for live and dead cells confirmed that the reduction in the biomass was achieved by a mechanism that did not affect bacterial viability, reducing the chances for the evolution of resistant strains.